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The study was carried out with the objective of evaluating the effects of using phytase levels at different energy densities in the diet of broilers, from 28 to 35 days of age. The experimental diets contained increasing levels of apparent metabolizable energy corrected for nitrogen balance and different levels of phytase. Growth performance variables, nutrient digestibility, as well as variables related to bone integrity were examined. Diets containing 500 FTU/kg in combination with 3150kcal.kg-1 of AMEn resulted in better growth performance. Supplementation with 500 and 1000 FTU/kg in the diets provide better coefficients of apparent and ileal digestibility of calcium and phosphorus, but negatively influence the deposition of calcium, phosphorus, dry matter, and ash in the tibia, in addition to adversely affecting the breakage and area of these bones in broilers. Bone length and the levels of magnesium and zinc present in the tibias were not influenced by the energy and phytase levels of the diets. There is no interaction between the different energetic densities and the phytase doses in the variables analyzed in the studied period. Increasing the energy density of diets resulted in improved apparent and ileal digestibility for most nutrients. RESUMO O estudo foi realizado com o objetivo de avaliar os efeitos da utilização de níveis de fitase em diferentes densidades energéticas na dieta de frangos de corte, no período de 28 a 35 dias de idade. As dietas experimentais contiveram níveis crescentes de energia metabolizável aparente corrigida para balanço de nitrogênio e diferentes níveis de fitase. Foram avaliadas variáveis de desempenho zootécnico, digestibilidade de nutrientes e variáveis relacionadas à integridade óssea. Dietas contendo 500 FTU/kg de fitase em combinação com 3150kcal.kg-1 de EMAn resultam em melhor desempenho zootécnico. A suplementação com 500 e 1000 FTU/kg nas dietas propicia melhores coeficientes de digestibilidade aparente e ileal do cálcio e do fósforo, mas influencia negativamente na deposição de cálcio, fósforo, matéria seca e cinzas das tíbias, além de afetar adversamente a quebra e a área desses ossos em frangos de corte. O comprimento do osso e os níveis de magnésio e zinco presentes nas tíbias não são influenciados pelos níveis energéticos e de fitase das dietas. Não há interação entre as diferentes densidades energéticas e as doses de fitase nas variáveis analisadas no período estudado. O aumento da densidade energética das dietas resulta em melhora na digestibilidade aparente e ileal para a maioria dos nutrientes.

8 Pags.- 1 Tabl.- 1 Fig., The influence of nine different Prunus rootstocks budded with the ‘Big Top’ nectarine cultivar was evaluated in a rootstock trial established at the Experimental Station of Aula Dei (Zaragoza, Spain). Chilling injury (CI) symptoms as browning, mealiness, leatheriness and bleeding were determined after 28 days of cold storage at 5°C plus 2 days of shelf-life. Agronomical basic fruit quality traits as firmness, soluble solids content (SSC) and titratable acidity (TA) were determined at harvest, after 28 days of fruit storage at 5°C and after 28 days plus 2 days of shelf-life. In addition, biochemical compounds as main individual sugars and organic acids, relative antioxidant capacity (RAC), total phenolics content (TPC), total flavonoids content (TFC), vitamin C (AsA) and anthocyanins content (AC) were also quantified. Moreover, enzymatic activity of key enzymes related with the tolerance against CI such as phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO) and peroxidase (POX) were determined. Significant effect of Prunus rootstocks was found on leatheriness and bleeding symptoms, firmness, TA, sugars and organic acids, AC and enzymatic activities. Sugars, organic acids, TPC, AC and the enzymatic activities of the phenylpropanoid pathway and ROS metabolism enzymes seemed to be important parameters to consider in the prevention of CI susceptibility. These results may be helpful in the choice of rootstocks inducing lower chilling injury susceptibility., This work was funded by the Spanish Ministry of Science Innovation and Universities (MICINN) grant RTI2018-094176-R-C33, cofounded by FEDER, and the Regional Gov. of Aragon (A44, T07_17R, A22_20R).

In this study, we focused on the fact that soil storage conditions in the laboratory have never been considered as a key factor potentially leading to high variation when measuring effects of microplastics on soil microbial activity. We stored field-collected soils under four different conditions [room-temperature storage, low-temperature storage (LS), air drying (AD), and heat drying] prior to the experiment. Each soil was treated with tire wear particles (TWPs), and soil microbial activities and water aggregate stability were investigated after soil incubation. As a result, microbial activities, including soil respiration and three enzyme activities (β-glucosidase, N-acetyl-β-glucosaminidase, and phosphatase), were shown to depend on soil storage conditions. Soil respiration rates increased with the addition of TWPs, and the differences from the control group (no TWPs added) were more pronounced in the AD TWP treatment than in soils stored under other conditions. In contrast, phosphatase activity followed an opposing trend after the addition of TWPs. The AD soil had higher phosphatase activity after the addition of TWPs, while the LS soil had a lower level than the control group. We suggest that microplastic effects in laboratory experiments can strongly depend on soil storage conditions.

The development of an effective, realistic, and sustainable microbial biorefinery depends on several factors, including as one of the key aspects an adequate selection of microbial strain. The oleaginous red yeast Rhodotorula sp. has been studied as one powerful source for a plethora of high added-value biomolecules, such as carotenoids, lipids, and enzymes. Although known for over a century, the use of Rhodotorula sp. as resource for valuable products has not yet commercialized. Current interests for Rhodotorula sp. yeast have sparked from its high nutritional versatility and ability to convert agro-food residues into added-value biomolecules, two attractive characteristics for designing new biorefineries. In addition, as for other yeast-based bioprocesses, the overall process sustainability can be maximized by a proper integration with subsequent downstream processing stages, for example, by using eco-friendly solvents for the recovery of intracellular products from yeast biomass. This review intends to reflect on the current state of the art of microbial bioprocesses using Rhodotorula species. Therefore, we will provide an analysis of bioproduction performance with some insights regarding downstream separation steps for the extraction of high added-value biomolecules (specifically using efficient and sustainable platforms), providing information regarding the potential applications of biomolecules produced by Rhodotorula sp, as well as detailing the strengths and limitations of yeast-based biorefinery approaches. Novel genetic engineering technologies are further discussed, indicating some directions on their possible use for maximizing the potential of Rhodotorula sp. as cell factories. Key points: • Rhodotorula sp. are valuable source of high value-added compounds. • Potential of employing Rhodotorula sp. in a multiple product biorefinery. • Future perspectives in the biorefining of Rhodotorula sp. were discussed.

Resistance responses in the leaves of eight rose genotypes, Knock Out (highly resistant), Arka Nishkant (moderately resistant), R. multiflora (highly susceptible), Arka Swadesh (highly susceptible), IIHRR 13-4 (susceptible), Arka Parimala (susceptible), R. indica (susceptible) and IIHRR 4-15-12 (moderately susceptible), exhibiting varied levels of resistance against black spot were investigated post artificial inoculation with black spot pathogen, Diplocarpon rosae. There was consistent increase in the activities of defense related enzymes such as catalase, peroxidase, polyphenol oxidase, superoxide dismutase and phenylalanine ammonia lyase and other defense related secondary metabolites like phenols and flavonoids at different phases of black spot progression and increase was high in resistant genotypes Knock Out and Arka Nishkant. The peak activity of defense enzymes and high concentration of other metabolites was witnessed during early stages of infection in the resistant genotypes while it was during later phase in the susceptible genotypes. These results suggested that the faster and stronger activation of defense system is associated with the resistance against black spot in the rose genotypes.

Naturally evolved enzymes, despite their astonishingly large variety and functional diversity, operate predominantly through thermochemical activation. Integrating prominent photocatalysis modes into proteins, such as triplet energy transfer, could create artificial photoenzymes that expand the scope of natural biocatalysis

Methionine and taurine are amino acids (AA) that are usually deficient when fish meal is replaced by plant proteins. In this study, three diets were tested in juvenile meagre (initial weight: 13.4 g) for 8 weeks. The D1 diet had 0.2% methionine and 1% taurine supplementation; the D2 and D3 diets had 0.6% methionine and 1% and 2% taurine supplementation, respectively. The results showed that meagre fed the D1 diet had lower specific growth rate (2.2 to 2.5), lower feed efficiency (0.9 to 1.2) and higher food conversion rate (FCR, 1.1 to 0.8) as well as a lower activity of the alanine aminotransferase (ALAT) enzyme. Furthermore, a higher recruitment of muscle fibres (46% compared to 36%) as well as a higher fibre density was observed (1019 compared to 870 fibres mm(-2)). This study shows that meagre requires a sufficient quantity of methionine in plant-based diets to avoid a reduction in fish performance. Furthermore, taurine supplementation in the D1 diet was not able to mitigate the effects of methionine deficiency. A higher taurine supplementation did not improve meagre performance. MAR02.01.01 FEAMP-0175 info:eu-repo/semantics/publishedVersion

There is an increasing interest in the upgrading of inexpensive and abundant C

The demand for novel, robust microbial biocatalysts for use in industrial and pharmaceutical applications continues to increase rapidly. As a result, there is a need to develop advanced tools and technologies to exploit the vast metabolic potential of unculturable microorganisms found in various environments. Single-cell and functional metagenomics studies can explore the enzymatic potential of entire microbial communities in a given environment without the need to culture the microorganisms. This approach has contributed substantially to the discovery of unique microbial genes for industrial and medical applications. Functional metagenomics involves the extraction of microbial DNA directly from environmental samples, constructing expression libraries comprising the entire microbial genome, and screening of the libraries for the presence of desired phenotypes. In this study, lipolytic enzymes from the Red Sea were targeted. A high-throughput single-cell microfluidic platform combined with a laser-based fluorescent screening bioassay was employed to discover new genes encoding lipolytic enzymes. Analysis of the metagenomic library led to the identification of three microbial genes encoding lipases based on their functional similarity and sequence homology to known lipases. The results demonstrated that microfluidics is a robust technology that can be used for screening in functional metagenomics. The results also indicate that the Red Sea is a promising, under-investigated source of new genes and gene products.

The aim of the present study was to establish the pathohistological and biochemical changes in ewes from the Laucane breed with ketosis. Blood samples were obtained from 106 dairy ewes for determination of β-hydroxybutyrate (ВНВА), non-esterified fatty acids (NEFA), glucose (Gl), aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT) and total bilirubin (Tb). The ewes were divided in three groups: pregnant, recently lambed and lactating. Target ewes were classified as healthy (C), affected with subclinical ketosis (SCK) and with clinical ketosis (CK) depending on their blood BHBA levels. The quantities of NEFA in sheep with SCK were statistically significantly elevated, while in sheep with CK – decreased. The levels of glucose decreased, while the activities of ASAT, ALAT and Tb levels were increased in ewes with SCK and CK ketosis. Histological examination revealed cellular vacuolation in hepatocytes, karyolysis, karyorrhexis, necrotic changes and high-grade fatty dystrophy of the liver and kidneys in ewes with ketosis., {"references":["Anoushepour, A., Mottaghian, P., Sakha, M. (2014). The comparison of some biochemical parameters in hyperketonemic and normal ewes. European journal of experimental biology, 4 (3), pp. 83– 87.","Aschenbach, J. R., Penner, G. B., Stumpff, F., Gäbel, G. (2011). Ruminant nutrition symposium: role of fermentation acid absorption in the regulation of ruminal pH. Journal of animal science, 89, pp. 1092–1107.","Balikci, E., Yildiz, A., Gurdogan, F. (2009). Investigation on some biochemical and clinical parameters for pregnancy toxemia in Akkaraman ewes. Journal of animal and veterinary advances, 8 (7), pp. 1268–1273.","Cal, L., Borteiro1, C., Benech, A., Rodas, E., Abreu, M. N., Cruz, J. C., González Monataña, R. (2009). Histological changes of the liver and metabolic correlates in ewes with pregnancy toxemia. Arquivo brasileiro de medicina veterinária e zootecnia, 61, pp. 306–312.","Clarkson, M. J. (2000). Pregnancy toxaemia. In: Diseases of sheep. Ed. W.B. Martin & I.D. Aitken, UK, pp. 315–317.","Djoković, R., Kurćubić, V., Ilić Z., Cincović, M., Petrović, M., Fratrić, N., Jašović, B. (2013). Evaluation of metabolic status in Simmental dairy cows during late pregnancy and early lactation. Veterinarsky arhiv, 83 (6), pp. 593–602","Dzhurov, A., Alexandrova, E., Alexandrov, M. (1989). Metodi za patohistologichni izsledvania. Zemizdat, Sofia, pp. 28–34. (in Bulgarian).","Feijó, J. O., Schneider, A., Schmitt, E., Brauner, C. C., Martins, C. F., Barbo-sa-Ferreira, M., Del Pino, F. A. B., Faria Junior, S. P., Rabassa, V. R., Corrêa, M. N. (2015). Prepartum administration of recombinant bovine somatotropin (rBST) on adaptation to subclinical ketosis of the ewes and performance of the lambs. Arquivo brasileiro de medicina veterinária e zootecnia, 67 (1), pp. 103– 108.","Ferris F. T., Herdson B. P., Dunnill S. M., Lee, M. R. (1969). Toxemia of Pregnancy in Sheep: a Clinical, Physiological, and Pathological Study. The journal of clinical investigation, 48, pp. 1643– 1655.","Garverick, H. A., Harris, M. N., Vogel-Bluel, R., Sampson, J. D., Bader, J., Lamberson, W. R., Spain, J. N., Lucy, M. C., Youngquist, R. S. (2013). Concentrations of nonesterified fatty acids and glucose in blood of periparturient dairy cows are indicative of pregnancy success at first insemination. Journal of dairy science, 96, pp. 181–188","Harmeyer, J., Schlumbohm, C. (2006). Pregnancy impairs ketone body dispo-sal in late gestation: Implications for onset of pregnancy toxaemia. Research in veterinary science, 81, pp. 254–264.","Hefnawy, A. E., Shousha, S., Youssef, S. (2011). Hematobiochemical profile of pregnant and experimentally pregnancy toxemic goats. Journal of basic and applied chemistry, 1, pp. 65–69","Holtenius, P., Holtenius, K. (1996). New aspects of ketone bodies in energy metabolism of dairy cows: a review. Journal of veterinary medicine series A, 43, pp. 579–587.","Kahn, C. M., Line, S. (2005). Pregnancy toxemia in Ewes. In: The Merck Veterinary Manual. 9th Ed., Merck & Co Whitehouse Station, pp. 828–830","Kaneko, J. J., Harvey, J. W., Bruss, M. L. (2008). Clinical biochemistry of domestic animals. 5th Ed., Academic Press, 3, 4 & appendix No.VIII.","Kuhla, B., Nurnberg, G., Albrecht, D., Görs, S., Hammon, H. M., Metges, C. C. (2011). Involvement of skeletal mus cle protein, glycogen, and fat metabolism in the adaptation on early lactation of dairy cows. Journal of proteome research, 10 (9), pp. 4252–62.","Marutsova, V., Binew, R. (2020). Changes in blood enzyme activities and some liver parameters in goats with subclinical ketosis. Bulgarian journal of veterinary medecine, 23 (1), pp. 70–79.","McCausland, I. P., O'Hara, P. J. (2004). Spontaneous toxaemia of pregnancy in sheep: A study of renal function and glomerular fine structure. Journal of comparative pathology, 84 (3), pp. 375–80.","Moallem, U., Rozov, A., Gootwine, E., Hoing, H. (2015). Plasma concentra-tions of key metabolites and insulin in late-pregnant ewes carrying 1 to 5 fetuses. Journal of animal science, 90 (1), pp. 318– 324.","Moghaddam, G. H., Hassanpour, A. (2008). Comparison of blood serum glucose, beta hydroxybutyric acid, blood urea nitrogen and calcium concentrations in pregnant and lambed ewes. Journal of animal and veterinary advances, 7, pp. 308–311.","Radostits, O. M., Blood, D. C., Henderson, J. A. (2007). Veterinary Medicine. 8th Ed., London: Bailliere & Tindall Publication Ltd, pp. 1450–1452.","Reece, W.O. (2004). Dukes' physiology of domestic animals. 12th Ed. Comstock publishing associates a division of Cornell university press.","Roche, J. R., Bell, A. W., Overton, T. R., Loor, J. J. (2013). Invited review: Nutritional management of the transition cow in the 21st century – a paradigm shift in thinking. Animal production science, 53 (9), pp. 1000–1023.","Schlumbohm, C., Harmeyer J. (2008). Twin-pregnancy increases susceptibility of ewes to hypoglycaemicstress and pregnancy toxaemia. Research in veterinary science, 84, pp. 286–299.","Sigh, S. K., Srivastav, C. P., Lonkar, P. S., Prasad, M. C. (1995). Pregnancy toxaemia in sheep: Retrospective and prospective studies. Indian journal of animal science, 65 (9), pp. 995–997","Simonov, M., Vlizlo, V. (2015). Some blood markers of the functional state of liver in dairy cows with clinical ketosis. Bulgarian journal of veterinary medecine, 18 (1), pp. 74–82.","Song, Y., Li, N., Gu, J., Fu, S., Peng, Z., Zhao, C., Zhang, Y., Li, X., Wang, Z., Li, X. Liu, G. (2016). β-Hydroxybutyrate induces bovine hepatocyte apoptosis via an ROS-p38 signaling pathway. Journal of dairy science, 99 (11), pp. 9184–9198","Sordillo, L. M., Raphael W. (2013). Significance of metabolic stress, lipid mobilization and inflammation on transition cow disorders. Veterinary clinics of North America: food animal practice, 29, pp. 267–278.","Van den Top, A. M., Van Tol, A., Jansen, H., Geelen, M. J. H., Beynen, A. C. (2005). Fatty liver in dairy cows post partum is associated with decreased concentration of plasma triacylglycerols and decreased activity of lipoprotein lipase in adipocytes. Journal of dairy research, 72, pp. 129–137.","Van Saun, R. J. (2000). Pregnancy toxemia in a flock of sheep. Journal of the American veterinary medical association, 217, pp. 1536–1539."]}

Different degrees in the biological activities of Canavalia rosea had been previously reported . In this study, our group assessed the cardioprotective effects of the ethyl acetate fraction (EAcF) of the Canavalia rosea leaves. Firstly, it was confirmed, by in vitro approach, that the EAcF has high antioxidant properties due to the presence of important secondary metabolites, as flavonoids. In order to explore their potential protector against cardiovascular disorders, hearts were previously perfused with EAcF (300 μg.mL-1) and submitted to the global ischemia followed by reperfusion in Langendorff system. The present findings have demonstrated that EAcF restored the left ventricular developed pressure and decreased the arrhythmias severity index. Furthermore, EAcF significantly increased the glutathiones peroxidase activity with decreased malondialdehyde and creatine kinase levels. EAcF was effective upon neither the superoxide dismutase, glutationes reductase nor the catalase activities. In addition, the Western blot analysis revealed that ischemia-reperfusion injury significantly upregulates caspase 3 protein expression, while EAcF abolishes this effect. These results provide evidence that the EAcF reestablishes the cardiac contractility and prevents arrhythmias; it is suggested that EAcF could be used to reduce injury caused by cardiac reperfusion. However more clinical studies should be performed, before applying it in the clinic.

Penaeus spp. are the most cultivated type of shrimp because they have rapid growth and good adaptation to farming conditions. Due to diseases that result in high mortality and a decreased product quality, the cultivation of these shrimp globally, and in Cuba in particular, comes with the risk of significant financial losses. This study examined the effect of fructooligosaccharide (FOS, 1-kestose) on the growth and immune response of shrimp, as well as the multitude of Vibrio spp. and Pseudomonas spp. in Penaeus vannamei post-larvae under culture conditions. Fructooligosaccharide was applied in a completely randomized manner at a concentration of 0.4%, in both experimental groups with seventeen tanks each. In the results of this investigation, animals of greater weight (control 6.8 ± 0.2 mg; FOS 9.5 ± 0.3 mg; p < 0.001), length (control 1.1 ± 0.1 mm; FOS 1.3 ± 0.1 mm; p < 0.001), and survival (control 61.7% (95% CI of median 54.2–70.0); FOS 76.6% (95% CI of median 72.1–84.2); p < 0.001) were obtained when administered FOS relative to the control. An increase in the activity of proteases (p < 0.001), enzymes of the innate immune system such as phenoloxidase (p < 0.001), and lysozymes (p < 0.001) was observed, as well as an increase in the number of lectins (p < 0.001). Changes in the microbiota could be observed, with a reduction in Vibrio spp. and Pseudomonas spp. (control 2.4 × 103 ± 0.5 × 103; FOS 1.1 × 102 ± 0.3 × 102; p < 0.001). FOS improves the quality of the post-larvae of P. vannamei as reflected in the length, weight, and survival of the animals. Moreover, FOS stimulates the P. vannamei immune system through the enzymatic activities of phenoloxidase, lysozyme, and a number of lectins. The reduction in the population load of Vibrio spp. and Pseudomonas spp. might be a consequence of the improvement in the quality and immune system of P. vannamei.

Sporothrix brasiliensis is the main agent of zoonotic sporotrichosis transmitted by domestic cats in South America. In humans, sporotrichosis commonly presents with cutaneous or lymphocutaneous lesions, and in cats, with multiple ulcerated skin lesions associated with enlarged lymph nodes and respiratory signs. Fungal virulence factors may affect the clinical presentation of the mycoses. Sporothrix spp. present some virulence factors. This study aims to compare 24 S. brasiliensis strains from 12 familiar outbreaks of cat-to-human transmitted sporotrichosis. Fungal growth in different substrates, thermotolerance, resistance to oxidative stress, and production of enzymes were evaluated. An invertebrate model of experimental infection was used to compare the virulence of the strains. The strains grew well on glucose and N-acetyl-D-glucosamine but poorly on lactate. Their thermotolerance was moderate to high. All strains were susceptible to hydrogen peroxide, and the majority produced hemolysins but not phospholipase and esterase. There was no significant difference in the putative virulence-associated factors studied among the different hosts. Moreover, strains isolated from a human and a cat from four familiar outbreaks presented a very similar profile of expression of these factors, reinforcing the zoonotic transmission of S. brasiliensis in Brazil and demonstrating the plasticity of this species in the production of virulence factors.

The treatment of saturated horse beds before they arrive at their final destination is necessary to avoid the risk of animal and environmental contamination. For this purpose, the composting process has great functionality due its to low cost, effectiveness, and operational ease. However, because of the nature of the materials used, this process can be long, and it is necessary to improve it to optimize composting cycles. This work aimed to isolate and identify fungi present in the compost piles of saturated equine bedding made with shavings and rice straw, identifying those with the greatest potential for cellulase production. Using specific cellulolytic media containing shavings or rice straw, seven strains were isolated. The total cellulase enzymatic activity of the isolates from the beds made with shavings was lower than that obtained from rice straw beds. Four strains showed high enzymatic potential for use in the shavings substrate (MA -6 2 f1, MA -6 2 f2, MA -7 9, and MA -7 10) and three for the rice straw substrate (PA -7 5, PA -7 7, and PA -7 10). The isolate PA -7 5 reached 0.376 IU mL−1, the best index among all the isolates. These isolates were identified as belonging to the Aspergillus fumigatus species.

Owing to their wide range of applications in the control of ticks and insects in horticulture, forestry, agriculture and food production, pyrethroids pose a significant threat to the environment, including a risk to human health. Hence, it is extremely important to gain a sound understanding of the response of plants and changes in the soil microbiome induced by permethrin. The purpose of this study has been to show the diversity of microorganisms, activity of soil enzymes and growth of Zea mays following the application of permethrin. This article presents the results of the identification of microorganisms with the NGS sequencing method, and of isolated colonies of microorganisms on selective microbiological substrates. Furthermore, the activity of several soil enzymes, such as dehydrogenases (Deh), urease (Ure), catalase (Cat), acid phosphatase (Pac), alkaline phosphatase (Pal), β-glucosidase (Glu) and arylsulfatase (Aryl), as well as the growth of Zea mays and its greenness indicators (SPAD), after 60 days of growth following the application of permethrin, were presented. The research results indicate that permethrin does not have a negative effect on the growth of plants. The metagenomic studies showed that the application of permethrin increases the abundance of Proteobacteria, but decreases the counts of Actinobacteria and Ascomycota. The application of permethrin raised to the highest degree the abundance of bacteria of the genera Cellulomonas, Kaistobacter, Pseudomonas, Rhodanobacter and fungi of the genera Penicillium, Humicola, Iodophanus, Meyerozyma. It has been determined that permethrin stimulates the multiplication of organotrophic bacteria and actinomycetes, decreases the counts of fungi and depresses the activity of all soil enzymes in unseeded soil. Zea mays is able to mitigate the effect of permethrin and can therefore be used as an effective phytoremediation plant.

International audience; This paper aims to investigate the effect of extrusion at high solid loading on corn stover (CS) properties and its enzymatic hydrolysis. This biomass was extruded under different screw speeds and different solid loadings and the impact of these parameters on physicochemical properties was evaluated. It was found that lignocellulosic components were not significantly affected by the pretreatment, while surface area increased with solid loading and rotation speed. Different enzyme cocktails were used for the enzymatic hydrolysis of extruded and untreated CS. Overall, mild twin-extrusion enhanced the enzymatic hydrolysis of corn stover through an increase in glucose and xylose yields by 134-212% and 214-294% respectively when using T. longibrachiatum cellulase. The highest sugar content was obtained from CS extruded under 400 g total solids (TS) per liter and 200 rpm. The energy efficiency of the pretreatment was also assessed and was found to be maximal at 400 gTS/L and 200 rpm.

Aims The aim of this study was to investigate the effectiveness of bread as substrate for γ-aminobutyric acid (GABA) biosynthesis, establishing a valorization strategy for surplus bread, repurposing it within the food chain. Methods and Results Surplus bread was fermented by lactic acid bacteria (LAB) to produce GABA. Pediococcus pentosaceus F01, Levilactobacillus brevis MRS4, Lactiplantibacillus plantarum H64 and C48 were selected among 33 LAB strains for the ability to synthesize GABA. Four fermentation experiments were set up using surplus bread as such, added of amylolytic and proteolytic enzymes, modifying the pH or mixed with wheat bran. Enzyme-treated slurries led to the release of glucose (up to 20 mg g−1) and free amino acid, whereas the addition of wheat bran (30% of bread weight) yielded the highest GABA content (circa 800 mg kg−1 of dry weight) and was the most suitable substrate for LAB growth. The selected slurry was ultimately used as an ingredient in bread making causing an increase in free amino acids. Conclusions Besides the high GABA concentration (148 mg kg−1 dough), the experimental bread developed in this study was characterized by good nutritional properties, highlighting the efficacy of tailored bioprocessing technologies as means to mitigate food wastage. Significance and Impact of Study Our results represent a proof of concept of effective strategies to repurpose food industry side streams.

Mitochondrial iron transporter (MIT) genes are essential for mitochondrial acquisition/import of iron and vital to proper functioning of mitochondria. Unlike other organisms, research on the MITs in plants is limited. The present study provides comparative bioinformatics assays for the potato MIT gene (StMIT) as well as gene expression analyses. The phylogenetic analyses revealed monocots-dicot divergence in MIT proteins and it was also found clade specific motif diversity. In addition, docking analyses indicated that Asp172 and Gly100 residues to be identified as the closest residues binding to ferrous iron. The percentage of structure overlap of the StMIT 3D protein model with Arabidopsis, maize and rice MIT proteins was found between 80.18% and 85.71%. The transcript analyses exhibited that the expression of StMIT was triggered under drought and salinity stresses. The findings of the present study would provide valuable leads for further studies targeting specifically the MIT gene and generally the plant iron metabolism.

Gram-positive bacteria are some of the earliest known life forms, diverging from gram-negative bacteria 2 billion years ago. These organisms utilize sortase enzymes to attach proteins to their peptidoglycan cell wall, a structural feature that distinguishes the two types of bacteria. The transpeptidase activity of sortases make them an important tool in protein engineering applications, e.g., in sortase-mediated ligations or sortagging. However, due to relatively low catalytic efficiency, there are ongoing efforts to create better sortase variants for these uses. Here, we use bioinformatics tools, principal component analysis and ancestral sequence reconstruction, in combination with protein biochemistry, to analyze natural sequence variation in these enzymes. Principal component analysis on the sortase superfamily distinguishes previously described classes and identifies regions of relatively high sequence variation in structurally-conserved loops within each sortase family, including those near the active site. Using ancestral sequence reconstruction, we determined sequences of ancestral Staphylococcus and Streptococcus Class A sortase proteins. Enzyme assays revealed that the ancestral Streptococcus enzyme is relatively active and shares similar sequence variation with other Class A Streptococcus sortases. Taken together, we highlight how natural sequence variation can be utilized to investigate this important protein family, arguing that these and similar techniques may be used to discover or design sortases with increased catalytic efficiency and/or selectivity for sortase-mediated ligation experiments.

Active-site loops play essential roles in various catalytically important enzyme properties like activity, selectivity, and substrate scope. However, their high flexibility and diversity makes them challenging to incorporate into rational enzyme engineering strategies. Here, we report the engineering of hot-spots in loops of the cumene dioxygenase from Pseudomonas fluorescens IP01 with high impact on activity, regio- and enantioselectivity. Libraries based on alanine scan, sequence alignments, and deletions along with a novel insertion approach result in up to 16-fold increases in activity and the formation of novel products and enantiomers. CAVER analysis suggests possible increases in the active pocket volume and formation of new active-site tunnels, suggesting additional degrees of freedom of the substrate in the pocket. The combination of identified hot-spots with the Linker In Loop Insertion approach proves to be a valuable addition to future loop engineering approaches for enhanced biocatalysts., Bundesministerium für Bildung und Forschung, Projekt DEAL

Wheat is the cereal most susceptible to lodging, particularly during the flowering period and at the early ripening stage. The use of plant growth retardants (PGRs) is especially recommended when intensive nitrogen (N) fertilisation is applied, which increases the susceptibility of plants to lodging. This paper presents the results of tests into the effects of PGRs (PGR0—control; PGR1—chlormequat chloride (CCC) + trinexapac-ethyl (TE); PGR2—chlormequat chloride (CCC) + ethephon (ET)), and N dose—N0, N20, N40, and N60 [0, 20, 40, and 60 kg N ha−1] on the content of selected plant secondary metabolites (PSM) in the Indian dwarf wheat (Triticum sphaerococcum Percival) of the Trispa cultivar, and on the abundance of insect pests. In the developmental stage of wheat (BBCH 39), insects were collected with an entomological net. The study also investigated the effect of experimental factors on the physicobiochemical properties of the soil (pH in KC, granulometric composition, total organic carbon TOC, total nitrogen TN, fractional composition of humus, and the activity of enzymes). An increase in the plant secondary metabolite (PSM) and FRAP (ferring reducing ability of plasma) contents following the application of PGRs and N fertilisation already from as low a rate as 20 kg ha−1 was demonstrated. A significant positive correlation was noted between the abundance of Oulema spp. and the contents of total polyphenols, chlorogenic acid, and FRAP. No such relationship was noted for Aphididae or Thysanoptera. TOC content was higher on the plots on which N fertilisation was applied at the highest rate and after the application of PGRs. The factor determining the TN content was N fertilisation. Soil samples of the PGR0 N0 treatment were characterised by the greatest proportion of carbon in the humic and fulvic acid fractions and by the smallest proportion of carbon in the humin fraction. N fertilisation increased the proportion of carbon in the humin fraction on the plots on which no PGRs were applied. The study demonstrated an increase in the activity of oxidoreductive enzymes following the application of higher N rates. The application of PGRs resulted in no inhibition of enzymes in the soil compared to the control (PGRs0).

The distribution of primary and secondary metabolites within a fruit can affect its nutritional and organoleptic quality, as fruit can vary in size and shape. This study investigated the differences in the distribution of the primary and secondary metabolites in the fruit of four strawberry cultivars (‘Asia’, ‘Clery’, ‘Frederica’, and ‘Sandra’) that were collected at one harvest point. The study included an analysis of the individual sugars, organic acids, phenolic compounds, and enzymes responsible for the degradation of phenolics. All of the studied cultivars showed a lower pH, higher total organic acid content, and lower glucose and fructose content in the outer part of the fruit. Differences were also observed in the total phenolic and anthocyanin contents, which were always higher in the outer part. The absolute differences in the total phenolic content ranged from 3723 to 6154 mg kg$_{-1}$ dry weight. Our results provide a basis for understanding the differences in the biosynthesis of these metabolites within this fruit and prove that it is essential to mix samples well before extractions to obtain results that are representative of the whole fruit. Nasl. z nasl. zaslona. Opis vira z dne 22. 5. 2023. Bibliografija: str. 12-13. Abstract.

An experiment was conducted to evaluate the effect of diets with reduced energy level content, supplemented with carbohydrase, on broiler performance and the coefficient of metabolizability of nutrients. A total of 720 one-day-old male Cobb-500 chicks were distributed in a completely randomized design, with six treatments, eight repetitions of 15 birds each. The treatments were: (1) a positive control, basal diet to meet the requirements of broiler chickens (PC); (2) a negative control, basal diet with a reduction of 80 kcal kg-1 (NC); (3) NC + alphagalactosydase; (4) NC + xylanase; (5) NC + xylanase and alphagalactosydase, and (6) NC + enzymatic blend (alphagalactosydase, xylanase, pectinase and amylase). The nutrient digestibility was not improved by use of enzymes. At 7 days of age, the broilers which were fed diets supplemented with enzymes showed a lower feed intake (FI) and better feed conversion ratio (FCR) than the broilers fed on PC. Both the NC and enzymatic blend resulted in a worse performance of the broilers at 21, 35 and 42 days old. The use of alphagalactosydase and xylanase, isolated or in combination, in a corn-soybean meal-based diet is effective in improving the growth performance of broilers fed energy-reduced diets.

Lysosomal storage disease is caused by the deficiency of a single hydrolase (lysosomal enzymes). GM2 gangliosidoses are autosomal recessive disorders caused by deficiency of β-hexosaminidase and Tay-Sachs disease (TSD) is one of its three forms. Methods: To perform a review of the state of the art on TSD and describe its definition, epidemiology, etiology, physiopathology, clinical manifestations, as well as advances regarding its diagnosis and treatment. A literature search was carried out in PubMed using the MeSH term “Tay-Sachs Disease”. Results: after the initial search was conducted, 1 233 results were retrieved, of which 53 articles were finally selected. TSD is caused by the deficiency of the lysosomal enzyme β-hexosaminidase A (HexA) and is characterized by neurodevelopmental regression, hypotonia, hyperacusis, and cherry-red spots in the macula. Research on molecular pathogenesis and the development of possible treatments has been limited, consequently, there is no treatment established to date. Conclusions: TSD is an autosomal recessive neurodegenerative disorder. Death usually occurs before the age of five. More research and studies on this type of gangliosidosis are needed to find adequate treatment. Trial registration: The study was registered at https://www.chictr.org.cn (ChiCTR1800019663). Keywords: Tay-Sachs Disease; Gangliosidoses; Hexosaminidase A; Enzymes; Jews (MeSH).

Global warming has been raising concerns for the state of the climate and the resulting adverse effects, due to the rise in levels of greenhouse gases including methane, nitrous oxide and carbon dioxide. As the atmospheric concentration of carbon dioxide has been rapidly increasing each year, it is pertinent to work on the decrease of such levels through the development of renewable methods of electricity generation and development of efficient methods of conversion of CO2 to chemicals. Electrochemical conversion of CO2 into reduced C1 compounds is a useful technique to reduce carbon dioxide levels as it is easy to select the appropriate reaction conditions. As electrochemical reduction of CO2 is associated with low product selectivity and efficiency due to competition with hydrogen evolution reactions, enzymatic electroreduction of carbon dioxide can be utilized due to the high selectivity of enzymes. This thesis demonstrates novel approaches to the direct immobilization of metal-independent formate dehydrogenases on electrode surfaces to facilitate the direct catalysis of carbon dioxide reduction to formate, without the need for NADH/NAD+ cofactors. 2024-05-23

Plastic pollution is not inevitable. All the players in the food packaging industry are mobilizing to find alternatives to packaging made from petroleum products while maintaining its primary function of protecting the food. In this context, materials such as nanofibrillated cellulose, alginate and chitosan, which are natural, renewable and biodegradable resources, are showing their full interest and are the subject of much research. These polymers are compatible with each other and make it possible to obtain composite biomaterials with better mechanical, thermal and barrier properties than those used independently. . In this project, the objectives are to compare the water and oxygen barrier properties of films composed of NFC alginate, NFC chitosan and NFC alginate chitosan, by combining different percentages of these three polymers. This study will be completed by measuring the affinity of the lacasse enzyme with the NCF alginate and NCF chitosan films. The protocols used to manufacture the films and the proportions are based on those carried out by Valentina Roxana Birsan. The Moister Content, water solubility, water uptake tests and the angle measurement of a drop of water on the films using the goniometer show that the NCF + Chitosan films from 50% have a behavior hydrophobic whereas NCF + Alginate films are hydrophilic. The analysis of the polarity and the dispersion values of the films highlights that the NCF + Alginate films obtain better results for the oxygen barrier properties compared to the films composed of NCF + Chitosan. Films composed of 50% NFC +25% alginate +25% chitosan improve water barrier properties. Measurement of the oxygen barrier properties at 0% humidity showed that NCF + Alginate films are oxygen impermeable due to their high polarity. At 50% humidity, NCF + Alginate films show an overall decrease in oxygen barrier properties. However, the measurement of this barrier property for NCF + Chitosan films needs to be completed to have conclusive results. Finally, the evaluation of the affinity to the laccase enzyme of the NCF + Alginate films and of the NCF + Chitosan films was carried out by measuring the oxidation of laccase by ABTS with a spectrophotometer. For the films composed of NCF + Chitosan, only the films with 30% chitosan reveal an affinity to this enzyme. These results would require further investigation. Films with alginate show an affinity to laccase proportional to the amount of alginate they contain. The production of packaging films composed of these three polymers and in particular in the proportions of 50% NCF + 25% alginate + 25% chitosan, seems to be a promising avenue. A complementary study on the affinity of the laccase enzyme on these composite films as well as their oxygen barrier properties is necessary. Incoming

The study evaluated the effect of substrate concentration and temperature on the production of amylase enzyme by Enterococcus faecalis from the wastes dump soil Sokoto Metropolis, Nigeria. Five bacteria species were isolated and biochemically characterized which include E. coli, B. subtilis, S. aureus, E. faecalis and P. aeroginosa. The screening of identified bacteria for amylase production was carried out and all bacteria produced a wide halo on a starch agar. E. faecalis produced highest zone of clearance with 70mm,B.subtilis 50mm,S.aureus 34mm, E. coli 29mm,thenB.cereus with the least zone of clearance of 24mm. proximate analysis of the potato peels sample shows that the potatoes peels has high carbohydrate content with value of 75.737 recorded. The effect of substrate concentration was studied at 5g,10g,15g and 20g. The highest amylase production was recorded (20g) with value of 1.218 and 15g with the least absorbance value of 0.973.The effect of temperature were studied at 25˚C, 35˚C, 45˚C, and 55˚C. The temperature of 45˚C has the highest absorbance value of 1.691. While the least was recorded at55˚C with value of 0.970.These shows that temperature and substrate concentration has a vital role to play in the production of microbial enzymes. It’s concluded in this study that waste dump site is a reservoir of important microorganisms and potato peels is a good substrate for enzyme production and therefore can be utilized instead of dumping it for natural degradation.

During the last decade, research into genetic markers in the casein gene cluster has been actively introduced in cattle breeding programs. A special interest has been paid to the polymorphism of the CSN3 gene, responsible for the expression of the k-casein, playing a key role in protein coagulation, interaction with whey proteins, stabilization, and aggregation of casein micelles. This paper aimed to determine the effect of CSN3 genetic polymorphism on acid; rennet; acid–rennet; heat- and acid-induced as well as heat- and calcium-induced coagulation in skimmed milk; and protein-standardized milk systems (UF, NF, RO, VE). The influence of polymorphic variants of the CSN3 gene on the coagulation ability of milk proteins was assessed by the particle size of casein micelles, protein retention factor in the clot, and coagulation ability (duration of induction period, mass coagulation period, dynamic viscosity in gel point). The correlation between CSN3 gene polymorphism and protein coagulation was revealed. Milk systems obtained from CSN3 BB milk were found to have the shortest duration of coagulation, formation of better gel strength values, and increased yield compared to CSN3 AA. This study will improve the efficiency of milk processing and optimize the technology of dairy product production.

Biofilm formation on abiotic surfaces has become a major public health concern because of the serious problems they can cause in various fields. Biofilm cells are extremely resistant to stressful conditions, because of their complex structure impedes antimicrobial penetration to deep-seated cells. The increased resistance of biofilm to currently applied control strategies underscores the urgent need for new alternative and/or supplemental eradication approaches. The combination of two or more methods, known as Hurdle technology, offers an excellent option for the highly effective control of biofilms. In this perspective, the use of functional enzymes combined with biosourced antimicrobial such as essential oil (EO) is a promising alternative anti-biofilm approach. However, these natural antibiofilm agents can be damaged by severe environmental conditions and lose their activity. The microencapsulation of enzymes and EOs is a promising new technology for enhancing their stability and improving their biological activity. This review article highlights the problems related to biofilm in various fields, and the use of encapsulated enzymes with essential oils as antibiofilm agents. KEY POINTS: • Problems associated with biofilms in the food and medical sectors and their subsequent risks on health and food quality. • Hurdle technology using enzymes and essential oils is a promising strategy for an efficient biofilms control. • The microencapsulation of enzymes and essential oils ensures their stability and improves their biological activities.

One of the main environmental stresses that hinder crop development as well as yield is salt stress, while the use of signal molecules such as calcium (Ca) has a substantial impact on reducing the detrimental effects of salt on different crop types. Therefore, a factorial pot experiment in a completely randomized design was conducted to examine the beneficial role of Ca (0, 2.5, and 5 mM) in promoting the physiological, biochemical, and growth traits of the wheat plant under three salt conditions viz. 0, 30, and 60 mM NaCl. Foliar application of Ca increased the growth of salt-stressed wheat plants through increasing photosynthetic pigments, IAA, proline, and total soluble sugars contents and improving antioxidant enzymes in addition to non-enzymatic antioxidants glutathione, phenol and flavonoids, β-carotene, and lycopene contents, thus causing decreases in the over-accumulation of free radicals (ROS). The application of Ca increased the activity of antioxidant enzymes in wheat plants such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), which scavenge reactive oxygen species (ROS) and relieved salt stress. An additional salt tolerance mechanism by Ca increases the non-antioxidant activity of plants by accumulating osmolytes such as free amino acids, proline, and total soluble sugar, which maintain the osmotic adjustment of plants under salinity stress. Exogenous Ca application is a successful method for increasing wheat plants’ ability to withstand salt stress, and it has a considerable impact on the growth of wheat under salt stress.

A reliance on fossil fuel has led to the increased emission of greenhouse gases (GHGs). The excessive consumption of raw materials today makes the search for sustainable resources more pressing than ever. Technical lignins are mainly used in low-value applications such as heat and electricity generation. Green enzyme-based modifications of technical lignin have generated a number of functional lignin-based polymers, fillers, coatings, and many other applications and materials. These bio-modified technical lignins often display similar properties in terms of their durability and elasticity as fossil-based materials while also being biodegradable. Therefore, it is possible to replace a wide range of environmentally damaging materials with lignin-based ones. By researching publications from the last 20 years focusing on the latest findings utilizing databases, a comprehensive collection on this topic was crafted. This review summarizes the recent progress made in enzymatically modifying technical lignins utilizing laccases, peroxidases, and lipases. The underlying enzymatic reaction mechanisms and processes are being elucidated and the application possibilities discussed. In addition, the environmental assessment of novel technical lignin-based products as well as the developments, opportunities, and challenges are highlighted.

The immobilization of enzymes on solid supports is an important challenge in biotechnology and biomedicine. In contrast to other methods, enzyme deposition in polymer brushes offers the benefit of high protein loading that preserves enzymatic activity in part due to the hydrated 3D environment that is available within the brush structure. The authors equipped planar and colloidal silica surfaces with poly(2-(diethylamino)ethyl methacrylate)-based brushes to immobilize Thermoplasma acidophilum histidine ammonia lyase, and analyzed the amount and activity of the immobilized enzyme. The poly(2-(diethylamino)ethyl methacrylate) brushes are attached to the solid silica supports either via a "grafting-to" or a "grafting-from" method. It is found that the grafting-from method results in higher amounts of deposited polymer and, consequently, higher amounts of Thermoplasma acidophilum histidine ammonia lyase. All polymer brush-modified surfaces show preserved catalytic activity of the deposited Thermoplasma acidophilum histidine ammonia lyase. However, immobilizing the enzyme in polymer brushes using the grafting-from method resulted in twice the enzymatic activity from the grafting-to approach, illustrating a successful enzyme deposition on a solid support.

Digestion is a fundamentally important process for an individual's life. However, the physical process of digestion is hidden inside the body, making it challenging to understand and a particularly difficult topic for students to learn in the classroom. Traditional approaches to teaching body processes include a mixture of textbook teaching and visual learning. However, digestion is not particularly visual. This activity is designed to engage students using a combination of visual, inquiry-based, and experiential learning approaches and introduces the scientific method to students in secondary school. The laboratory simulates digestion, creating a "stomach" inside of a clear vial. Students fill the vials with a protease solution and visually observe the digestion of food. By making predictions about the types of biomolecules that will be digested, students begin to learn and understand basic biochemistry in a relatable context, while simultaneously understanding anatomical and physiological concepts. We trialled this activity at two schools, where we received positive feedback from teachers and students, indicating that the practical enhanced student understanding of the digestion process. We see this lab as a valuable learning activity that can be extended broadly across multiple classrooms around the world.

Η παρούσα διδακτορική διατριβή στοχεύει στην παραγωγή βιομορίων και υψηλής αξίας προϊόντων από μικροοργανισμούς και πιο συγκεκριμένα από κυανοβακτήρια. Το στέλεχος του κυανοβακτηρίου που χρησιμοποιήθηκε στην παρούσα δουλειά έχει απομονωθεί από το επιβαρυμένο οικοσύστημα της λίμνης Παμβώτιδας. Τα παραγόμενα βιομόρια και προϊόντα είναι ένζυμα, λίπη και νανουλικά, που βρίσκουν εφαρμογές στους τομείς της βιομηχανίας, της ενέργειας και της ιατρικής. Τα ιδιαίτερα χαρακτηριστικά των κυανοβακτηριακών κυττάρων, όπως η ικανότητα ανάπτυξής τους σε μια πληθώρα από διαφορετικά ενδιαιτήματα, χρησιμοποιώντας ακόμα και λύματα, καθώς και η ικανότητά τους να παράγουν υψηλής αξίας προϊόντα την ίδια στιγμή, έχουν προσεγγίσει το ενδιαφέρον της ερευνητικής κοινότητας. Στην παρούσα μελέτη, μελετήθηκε ένα νέο στέλεχος κυανοβακτηρίου, απομονωμένου από τη λίμνη Παμβώτιδα. Η φυλογενετική του ανάλυση, το κατέταξε ως ένα στέλεχος Pseudanabaena sp., το οποίο ονοματίστηκε ως Pamv7. Τα κύτταρά του έχουν την ικανότητα να μεγαλώνουν τόσο σε αυτοτροφικές, όσο και ετεροτροφικές συνθήκες. Στην δεύτερη περίπτωση, μπορούν να μεγαλώσουν σε πληθώρα διαφορετικών υποστρωμάτων, αλλά όταν χρησιμοποιείται σελλοβιόζη, έχουν την ικανότητα να παράγουν β-γλυκοζιδάση. Το παρόν ένζυμο βρίσκει πληθώρα εφαρμογών στη βιομηχανία τροφίμων, αλλά και χαρτιού, χυμών, κρασιού κλπ, ενώ δεν έχει παραχθεί κατά το παρελθόν από το συγκεκριμένο τύπο μικροοργανισμών. Έτσι, μετά το φυλογενετικό χαρακτηρισμό του Pamv7 και της βιομηχανική του ανάλυσης, έλαβε χώρα μια αναλυτική μελέτη του ενζύμου. Αφού εξακριβώθηκε η παραγωγή του ενζύμου και της δράσης του, έλαβε χώρα η αριστοποίηση των συνθηκών ανάπτυξης (συγκέντρωση υποστρώματος, επιλογή πηγής αζώτου και συγκέντρωσής της, θερμοκρασία) του μικροοργανισμού και της παραγωγής του ενζύμου, με στόχο την αναβάθμιση της παραγωγής (scale-up). Χρησιμοποιώντας τις άριστες συνθήκες ανάπτυξης, έλαβε χώρα μεγαλύτερη παραγωγή του ενζύμου σε σύστημα βιοαντιδραστήρα 2L. Εν συνεχεία, πραγματοποιήθηκε ο καθαρισμός της παραγόμενης β-γλυκοζιδάσης και προσδιορίστηκε το μοριακό της βάρος και τα θερμοδυναμικά της χαρακτηριστικά. Το ένζυμο με μοριακό βάρος 66 kDa και ενζυμική ενεργότητα 40U/L, με βέλτιστη θερμοκρασία δράσης και pH τους 40oCκαι 6.0 αντίστοιχα, εμφανίζεται ιδιαιτέρως σταθερό στο εύρος του όξινου0ουδέτερου pHs (6.0-7.0), ιδιότητα που το κάνει ιδιαιτέρως χρήσιμο για χρήση στη βιομηχανία. Καθώς τα Pseudanabaena sp. κύτταρα είχαν την ικανότητα να παράγουν β-γλυκοζιδάση, γεννήθηκε το ερώτημα του αν θα μπορούσαν να παράγουν και άλλα ένζυμα, όπως η α-γλυκοζιδάση. Έτσι, χρησιμοποιώντας μαλτόζη ως πηγή άνθρακα, παράχθηκε α-γλυκοζιδάση με ενζυμική ενεργότητα περίπου 1800 U/L. Καθώς και η α-γλυκοζιδάση εμφανίζει πληθώρα εφαρμογών, η μελέτη συνεχίστηκε ως προς την αριστοποίηση των συνθηκών παραγωγής της. Στη συνέχεια, χρησιμοποιώντας αυτές τις συνθήκες, έγινε η παραγωγή μεγάλης ποσότητας του ενζύμου σε σύστημα βιοαντιδραστήρα 2L, ο καθαρισμό του και η μελάτη των κινητικών και θερμοδυναμικών του χαρακτηριστικών. Η παραγόμενη α-γλυκοζιδάση είχε μοριακό βάρος 52 kDa, άριστες συνθήκες δράσης τους 40oC και το pH 7.0, αλλά σταθερότητα στην όξινη κλίμακα του pH (4.0-6.0), κάτι που την κάνει κατάλληλη για διεργασίες όπου χρησιμοποιούνται τέτοιες συνθήκες. Εκτός των παραπάνω, τα μικροφύκη και τα κυανοβακτήρια αποτελούν καλούς παραγωγούς λιπιδίων. Επιπλέον η περιβαλλοντική μόλυνση και ιδιαιτέρως το φαινόμενο του θερμοκηπίου, καθώς και η μείωση των φυσικών πόρων, έχει οδηγήσει στην αναζήτηση νέων, ανανεώσιμων μορφών ενέργειας, όπως το βιοντίζελ. Το γεγονός αυτό, σε συνδυασμό με την ικανότητα των κυανοβακτηρίων να χρησιμοποιούν ακόμα και λύματα ως υποστρώματα για την ανάπτυξή τους, έθεσε τα θεμέλια για την εφαρμογή ενός σχεδίου βιο-επεξεργασίας και βιο-οικονομίας, που μειώνει το περιβαλλοντικό αποτύπωμα, ενώ ταυτοχρόνως οδηγεί στην παραγωγή βιοντίζελ . Έτσι στην παρούσα διατριβή, τα κύτταρα χρησιμοποίησαν υδρολύματα από δύο σκανδιναβικά φυτά (silverbirchandNorwayspruce) για την ανάπτυξή τους και την παραγωγή λιπιδίων και βιοκαυσίμου. Η ανάλυση των παραγόμενων λιπιδίων υπέδειξε ένα λιπιδιακό προφίλ κατάλληλο για την παραγωγή βιοκαυσίμου, ενώ η ανάλυση των ιδιοτήτων του παραγόμενου βιοκαυσίμου υπέδειξε ικανοποιητική ποιότητα για πιθανή χρήση του. Τέλος τα κύτταρα Pseudanabaena sp. χρησιμοποιήθηκαν ως ‘εργοστάσια’ για την παραγωγή πολύ μικρών νανοσωματιδίων μέσω μιας πράσινης συνθετικής διαδικασίας. Το υδατικό εκχύλισμα του κυανοβακτηρίου δρα ως αναγωγικός και σταθεροποιητικός παράγοντας για τα Ag+ του AgNO3 προς τον σχηματισμό νανοσωματιδίων Agoκαι AgO. Επιπροσθέτων οι βιολογικές ενώσεις δρουν ως προστατευτικοί παράγοντες για τα νανοσωματίδια, προσδίδοντας τους πολλαπλές λειτουργίες. Τα μορφολογικά και δομικά χαρακτηριστικά τους προσδιορίστηκαν με τη χρήση τεχνικών όπως φασματομετρίες FTIR και XPS, X-rayDiffraction, μικροσκοπίες AFM και TEM. Επιπλέον τα παραγόμενα νανοσωματίδια εμφάνισαν εξαιρετικά ισχυρές αντιμοκροβιακές ιδιότητες που σχετίζονται άμεσα και με το χρόνο και με τη δόση, έναντι Gram-negativeEscherichiacoliκαιGram- positiveCorynebacteriumglutamicumβακτηριακών στελεχών (15μg/mlAgNPs (δηλ 3 μg/mlAg+ οδηγεί σε βακτηριακή μείωση πληθυσμού της τάξης του 95%). Εκτός από τις εφαρμογές όπου γίνεται χρήση της αντιμικροβιακής τους ικανότητας, τα νανοσωματίδια αργύρου μπουρούν να βρουν εφαρμογή και σαν νανοδομές σε περιβαλλοντικές, βιολογικές ή ιατρικές εφαρμογές. Συμπερασματικά, τα κύτταρα του κυανοβακτηρίου μπορούν να χρησιμοποιηθούν ως ‘εργοστάσιο’ για την παραγωγή μια πληθώρας βιομορίων και προϊόντων. Η παρούσα εργασία αποτελεί την πρώτη λεπτομερή έρευνα που αφορά την παραγωγή ενζύμων από τον συγκεκριμένο τύπο μικροοργανισμού. Επιπλέον τα κύτταρα εκυπηρετούν την ιδέα της βιοοικονομίας, καταναλώνοντας υπολείμματα και παράγοντας βιώσιμα και φιλικά προς το περιβάλλον καύσιμα. Τέλος, χρησιμοποιήθηκαν ως εναλλακτική πλατφόρμα για την παραγωγή εξαιρετικά μικρών νανοσωματιδίων αργύρου, μέσω μιας διαδικασίας χαμηλού κόστους, γρήγορης και φιλικής προς το περιβάλλον. The current PhD thesis aims at producing biomolecules and high added value products from microorganisms and more specific from cyanobacteria. The used strain is isolated from lake Pamvotis in Greece, that constitute a burdened ecosystem. The produced molecules and products were enzymes, lipids and nanomaterials, that find applications on both industry, energy and medicine domains. The special characteristics of cyanobacterium cells like their ability to grow under a plethora of different habitants, using even wastes, and their ability to produce at the same time different high value products have attached considerable research interest. In this study, a novel cyanobacterium strain, which was isolated from Lake Pamvotis, was studied. After its phylogenetic analysis, it was as Pseudanabaenasp. strain, named Pamv7. Pseudanabaenasp. cells had the ability to grow both in autotrophic and heterotrophic conditions. At the second circumstance, they could grow at a plethora of different substrates, but when at cellobiose, they were producing β-glucosidase. β-glucosidase finds a plethora of application on feed, food, wine, textile, juice industries, and it was not produced before by this type of microorganisms. The current work aimed to this production. So, after the phylogenetic characterization of Pseudanabaena sp. and its biochemical composition, the analytic study of β-glucosidase took place. After the above verification of both enzyme production and its action, the purification of the cultural conditions took place. It constitutes an extremely important step; it is important the achievement of higher production for a scale up. These parameters are substrate concentration, nitrogen source and its concentration and temperature of culture. After the optimizing of them, a scale up of the culture at a bioreactor system of 2L took place. The produced β-glucosidase was purified and its molecular weight was determined, while its kinetic and thermodynamic characteristics were determined. The enzyme had a reduced molecular mass of 66 kDa and activity of about 40U/L, while it was optimally active at 40 oC and pH 6.0 and stable in acidic-neural pHs (6.0-7.0), something that make it applicable to many industrial uses. As Pseudanabaena sp cells were able to produce β-glucosidase, a question was born. Why not and other enzymes, like a-glucosidases? So, using maltose as carbon substrate for the heterotrophic cultivation of Pseudanabaenasp. cells, a-glucosidase was produced with an activity of about 1800 U/L. As a-glucosidase has also great application in industry, the optimization of cell cultural conditions for the greatest enzyme production took place. After that, a scale up of the culture at a bioreactor system of 2L took place, and the produced enzyme was purified and their kinetic and thermodynamic properties were determined. The purified enzyme appeared a reduced molecular mass of 52 kDa, while it was optimally active at 40oC and pH 7.0 and stable in acidic pHs (4.0-6.0), that make it appropriate for applications which demand acidic conditions. Except of enzymes, microalgae and cyanobacteria are known as lipid producers. Moreover, environmental pollution and especially the greenhouse gases, the decrease of fossil fuels in parallel with the population increase, has already lead to search of new and renewable source of energy, like biodiesel. This fact in combination with the fact that cyanobacteria are able to use of wastes as substrates for their growth, can lay the foundation for the adaption of a biorefinery concept, with reduction of environmental footprint, towards the production of biodiesel. So, in this study, Pseudanabaena sp. cells used pretreated hydrolysates from forest biomasses (silver birch and Norway spruce) towards the production of lipids and so as biodiesel. The lipid analysis showed a lipid profile suitable for biodiesel production, while the analysis of the properties of the produced after transesterification biodiesel showed a satisfy quality. Finally, Pseudanabaenasp.cells were used as the factory for the production of extra small silver nanoparticles via a green synthetic process. The aqua cyanobacterial extract acts as reductant and stabilizing agent for Ag+ of AgNO3 to form Ago and AgO nanoparticles. Moreover, its biological compounds act as capping factors for AgNPs creating in such way multiple functionalities. The morphological and structural characteristics of the produced nanoparticles have been characterized using FTIR and XPS spectroscopy, X-ray Diffraction, AFM and TEM microscopy. Further the Ag NPs have been evaluated for their antibacterial action against Gram-negative Escherichia coli and Gram- positive Corynebacterium glutamicum bacterial cells, presented an extremely time and dose dependent lethal effect (15μg/ml of AgNPs (that is 3 μg/ml of Ag+) are able to decrease the cell population about 95%). The silver nanoparticles except of their excellent bactericidal properties, could be also used as an excellent nanosupport for a series of environmental, biological and medical applications. As a conclusion, cyanobacterium cells can act as a bio-factory for a plethora of different bio-molecules and products. This work constitutes the first detailed investigation that concerns the production of enzymes from this type of microorganisms. Moreover, they cells were serving the biorefinery concept, consuming residues and producing sustainable and ecofriendly fuel. Finally, Pseudanabaena sp. cells were used as an alternative platform for the preparation of ultra-small silver nanoparticle, through a low-cost, rapid and eco-friendly synthetic procedure. 264 p.

29 pags., 4 figs., 3 tabs., 1 graf., Accumulated greenhouse gas emissions are expected to increase from 36.2 Giga-tons (Gt) to 60 Gt over the next three decades. The global surface temperature has increased by¿+¿1.09¿°C since 2001, and might increase by¿+¿2.2¿°C in 2100, +3.6¿°C in 2200 and +4.6¿°C in 2500. These emissions and temperature rises cannot be reduced in their entirety, but they can be lowered by using enzymes. Enzymes are proteins that catalyze biochemical reactions that make life possible since 3.8 billion years ago. Scientists have been able to "domesticate" them in such a way that enzymes, and their engineered variants, are now key players of the circular economy. With a world production of 117 Kilo-tons and a trade of 14.5 Billion-dollars, they have the potential to annually decrease CO2 emissions by 1 to 2.5 Billion-tons (Bt), the carbon demand to synthesise chemicals by 200 Million tons (Mt), the amount of chemicals by 90¿Mt, and the economic losses derived from global warming by 0.5%, while promoting biodiversity and our planet¿s health. Our success to increase these benefits will depend on better integration of enzymatic solutions in different sectors., This study was conducted under the auspices of the FuturEnzyme Project funded by the European Union’s Horizon 2020 Research and Innovation Programme under Grant Agreement No. 101000327. MF also acknowledges Grants PID2020-112758RB-I00, PDC2021-121534-I00, and TED2021-130544B-I00 from the MCIN/AEI/10.13039/501100011033 and the European Union (“NextGenerationEU/PRTR”).

La transición energética en el mundo ha impulsado la producción de los biocombustibles. Colombia lleva un gran camino recorrido al ser el principal productor de aceite de palma en Latinoamérica y el cuarto a nivel mundial. El aceite de palma un aceite de primera generación, es materia prima en la producción de biodiesel junto con un alcohol y el catalizador. Estas materias primas deben cumplir ciertas características fisicoquímicas para aumentar la conversión de triglicéridos, diglicéridos y monoglicéridos a biodiesel. Para reducir aún más el impacto ambiental la producción de biocombustibles está migrando a nuevas metodologías que evalúan utilizar aceites de primera, segunda, tercer y cuarta generación; alcoholes diferentes al metanol y etanol. Así mismo, el catalizador de tipo enzima inmovilizada es una tecnología que genera buenas conversiones según la selección de enzima para la reacción de transesterificación o esterificación. Lipasas de tipo bacteriano como la Pseudomona Cándida Antártida logra conversiones cercanas al 80% con aceite de palma usado (tercera generación), una conversión del 97% con aceite de semilla de algodón y metanol. Así mismo, los reactores han pasado de ser reactores Batch, CSTR, PFR o quimiostatos a reactores biológicos de lecho empacado - PBR, reactores de elevación por aire – ALR y reactor de columna de burbujeo – BCR, que disminuyen los esfuerzos cortantes manteniendo la integridad de las enzimas. El seguimiento y análisis de estas nuevas tecnologías se realizó con herramientas bibliométricas bajo la ecuación de búsqueda “biodiesel and reactor and enzyme or lipase and pubyear > 2011” generando una revisión de 256 publicaciones. The energy transition in the world has boosted the production of biofuels. Colombia has come a long way as it is the main producer of palm oil in Latin America and the second in worldwide. Palm oil, a first generation oil, is a raw material in the production of biodiesel together with an alcohol and the catalyst. These raw materials must meet certain physicochemical characteristics to increase the conversion of triglycerides, diglycerides and monoglycerides to biodiesel. To further reduce the environmental impact, the production of biofuels is migrating to new methodologies that evaluate the use of first, second, third and fourth generation oils; alcohols other than methanol and ethanol. Likewise, the immobilized enzyme catalyst type is a technology that generates good conversions depending on the selection of enzyme for the transesterification or esterification reaction. Bacterial-type lipases such as Pseudomonas Candida Antarctica achieve conversions close to 80% with used palm oil (third generation), a conversion of 97% with cottonseed oil and methanol. Likewise, the reactors have gone from being Batch, CSTR, PFR or chemostat reactors to packed bed biological reactors - PBR, air lift reactors - ALR and bubble column reactor - BCR, which reduce shear stresses while maintaining integrity. of the enzymes. The monitoring and analysis of these new technologies was carried out with bibliometric tools under the search equation "biodiesel and reactor and enzyme or lipase and pubyear > 2011" generating a review of 256 publications. http://www.ustabuca.edu.co/ustabmanga/presentacion Maestría

The Botrytis bunch mold, Botrytis cinerea pathogen is a necrotrophic ascomycete that infects hundreds of plant species, including hazelnut. B. cinerea produces toxins that induce cell wall degrading enzymes. In the current research work, we used eight hazelnut varieties and recorded their resistance levels in response to B. cinerea infection. Results showed that different varieties respond differently to B. cinerea infection. Disease index analysis revealed the resistance level of eight hazelnut varieties in the order of DW > OZ > L3 > PZ > XD1 > YZ > L1 > QX. Moreover, cell membrane permeability as well as the activities of cell wall degrading enzymes were measured. The increased level of cell wall degrading enzymes facilitates the fungal pathogens’ ability to colonize plants and cause infection. According to the results obtained through enzyme analysis, the hazelnut varieties L1 and QX, which were proved to be highly susceptible against B. cinerea, had the highest cell wall degrading enzyme production. DW and OZ, which were revealed to be resistant varieties through disease index data, also showed relatively lower activity of degrading enzymes as compared to other varieties. Our comparison analysis between the disease index and enzyme production confirms that disease occurrence and plant susceptibility strongly depend upon cell wall permeability. Our enzyme activity results validated the resistance order revealed by disease index assessment results (DW > OZ > L3 > PZ > XD1 > YZ > L1 > QX), and varieties DW and OZ were found to be the most resistant, while QX and L1 were found to be the most susceptible varieties against B. cinerea infection. Our study lays the foundation to further explore other factors involved in grey mold resistance in hazelnut.

Antiviral compounds are crucial to controlling the SARS-CoV-2 pandemic. Approved drugs have been tested for their efficacy against COVID-19, and new pharmaceuticals are being developed as a complementary tool to vaccines However, there are not any effective treatment against this disease yet. In this work, a cheap and fast purification method of natural tyrosinase from Agaricus bisporus fresh mushrooms was developed in order to evaluate the potential of this enzyme as a therapeutic protein by the inhibition of SARS-CoV-2 3CLpro protease activity in vitro. Tyrosinase showed a mild inhibition of 3CLpro of around 15%. Thus, different variants of this protein were synthesized through chemical modifications, covalently binding different tailormade glycans and peptides to the amino terminal groups of the protein. These new tyrosinase conjugates were purified and characterized by circular dichroism and fluorescence spectroscopy analyses, and their stability under different conditions. Then all these tyrosinase conjugates were tested in 3CLpro protease inhibition. From them, the conjugate between tyrosinase and dextranaspartic acid (6kDa) polymer showed the highest inhibition, with an IC50 of 2.5 µg/ml and IC90 of 5 µg/ml, results that highlight the potential use of modified tyrosinase as a therapeutic protein and opens the possibility of developing this and other enzymes as pharmaceutical drugs against diseases., This work was supported by the Spanish National Research Council (CSIC) (project CSIC PTI-Global Health SGL2103036 (J.M.P). This work was supported by Fundación hna (grant to A.V.-C. and O.A.).

Expression of recombinant enzymes in bacteria – principally E. coli - is an important method to generate high yields of industrially valuable proteins in short manufacturing timescales. While an extensive understanding of recombinant protein expression at industrial scale in E. coli has been achieved, challenges such as the formation of inclusion bodies, present limitations that inhibit streamlined manufacturing of recombinant enzymes. The lack of consistent workflows to address enzyme solubility issues often results in difficulties in producing high quantities of catalytically active protein. Commercial alcohol dehydrogenases are enzymes often produced in recombinant systems that are primarily employed in industrial chemical catalysis; the formation of inclusion bodies currently limits product yield of these enzymes. The primary aim of the work described in this thesis was to develop a robust strategy for heterologous expression of aggregate-prone industrially relevant alcohol dehydrogenases to improve their solubility in E. coli, thereby alleviating time-consuming troubleshooting steps in the future. A systematic literature analysis was first carried out to identify routinely adopted strategies used to minimise the likelihood of inclusion body formation in E. coli. This revealed that expression methods typically relied upon ad hoc, trial-and-error approaches with limited guarantees of success. This led to the emergence of the need to develop a systematic approach to handle this problem. Select identified strategies were then used to guide a more systematic experimental approach to cytosolic protein expression, which considered the impact of parameters such as plasmid design, strain variation, media formulation, and expression conditions. Protein expression at relatively high medium pH (pH = 9) led to a 4.47-fold and 5.46-fold improvement in soluble yield for two alcohol dehydrogenase models. The experimental design was further developed to improve the cellular density of the high pH system through fitness modifications to the E. coli strains of interest via adaptive laboratory evolution; the final evolved strain provided a further 4.15-fold and 4.87-fold improvement to the soluble yield of the system for the two alcohol dehydrogenase models. The evolved expression strain, E. coli ALE strain ‘D9’, showed key biological differences compared to the parental strain, E. coli BL21 (DE3). While E. coli BL21 (DE3) responded to protein expression at high pH by reducing/modifying its ribosomal profile and translational processes, the evolved strain reduced pathways related to energy intensive metabolic processes to maintain cytoplasmic pH in alkaline growth conditions. This evolved strain contained seven key mutations that conferred changes to cell envelope stress responses, morphology, and transcriptional/translational regulation that contributed to its fitness. The final E. coli expression system developed in this thesis ultimately improved the soluble yield of two model ‘difficult to express’ alcohol dehydrogenases. At the same time, the work exemplified an alternative view to protein expression that stresses the importance of understanding how E. coli biology, enzyme physicochemical properties, and fermentation conditions can affect the solubility of a DtE enzyme; furthermore, this work highlights the importance matching the requirements of an enzyme of interest with the characteristics of the expression host employed., Johnson Matthey Cambridge. Grant: NQAG/260

The functional state of enrichment cultures of the Eustigmatophycean strains Vischeria vischeri MZ–E3 and MZ–E4 after 25-day cultivation in the BBM medium was studied. The concentrations of chlorophyll a, total carotenoids, protein, vitamins A and E, fatty acid peroxidation product content, an antioxidant enzyme, and succinate dehydrogenase activity were measured. MZ–E3 succinate dehydrogenase activity was significantly higher by 2.21 times; the MZ–E4 strain had 2.94 times higher glutathione peroxidase activity. The MZ–E3 antioxidant activity index and the MZ–E3 unsaturation of fatty acids were 1.3 and 1.25 times higher than the MZ–E4. The retinol and α-tocopherol content of the MZ–E3 was 28.6% and 38.76% higher than MZ–E4. The main fatty acid profile differences were the 3.46-fold and 3.92-fold higher stearic and eicosapentaenoic acid content in the MZ–E4 biomass. MZ–E3 had higher antioxidant, energy, and metabolic and photosynthetic status than MZ–E4. The antioxidant status of the studied strains showed the dependence of the adaptive mechanisms of each, associated with differences in the ecological conditions of the biotopes from which they were isolated. These strains are promising for producing α-tocopherol and biomass enriched with omega-3 and omega-6 fatty acids.

Food industries have been exploited enzymes as green catalysts because enzymes from microbial, animal or plants origins offer unsurpassed physiochemical, biological and biocatalytic functionalities that allow biocatalysis with selectivity and specificity in converting raw materials. Enzyme-catalyzed processing has been used to extract bioactive substances, improve sensory, flavor and texture features, generate nutrient-rich foods, and eradicate food safety hazards. Enzymes give various benefits over traditional catalytic agents regarding process efficiency, sustainability, reusability, and cost-effective ratio. Over the years, some technologies have been used and optimized in order to improve biocatalysis for industrial applications. Multi-enzyme catalytic cascade based on a combination of enzymes offers diverse prospects in biosynthesis, biocatalysis, and biotransformation. Enzyme immobilization has allowed robustness and optimal functions in extreme industrial environments (e.g. high temperature) with no activity loss. Besides that, protein engineering has been provided exceptional tools for designing and producing novel biocatalysts with improved properties at industrial-scale food processing or green and sustainable chemical manufacturing. The progress in the random mutagenesis, screening, selection and computational project augmented the versatility and the rapid development of enzymes. Therefore, this chapter focuses on the emerging enzyme engineering trends and approaches to construct innovative biocatalysts with enhanced catalytic activity and highlights the food industry's recent application. The challenges and opportunities in developing biocatalytic procedures are also explored.

Colocalization of cascade enzymes is broadly discussed as a phenomenon that can boost the cascade reaction throughput, although a direct experimental verification is often challenging. This is mainly due to difficulties in establishing proper size regimes and in the analytical quantification of colocalization effect with adequate experimental systems and simulations. In this study, by taking advantage of reversible DNA-directed colocalization of enzymes on microspheres, we established a cascade system that can be used to directly evaluate the colocalization effect with exactly the same experimental settings except for the state of enzyme dispersion. In the regime of highly dilute microspheres of particular sizes, the colocalized cascade shows enhanced activity compared with the freely diffusing cascade, as evidenced by a shortened lag phase in the time-course production. Reaction-diffusion modeling reveals that the enhancement can be ascribed to the initial accumulation of intermediate substrate around the colocalized enzymes and is found to be carrier-size-dependent. This work demonstrates the dependence of the colocalization effect of enzyme cascades on an interplay of nano- and microscales, lending theoretical support to the rational design of highly efficient multienzyme catalysts.

Plants produce a broad variety of specialized metabolites with distinct biological activities and potential applications. Despite this potential, most biosynthetic pathways governing specialized metabolite production remain largely unresolved across the plant kingdom. The rapid advancement of genetics and biochemical tools has enhanced our ability to identify plant specialized metabolic pathways. Further advancements in transgenic technology and synthetic biology approaches have extended this to a desire to design new pathways or move existing pathways into new systems to address long-running difficulties in crop systems. This includes improving abiotic and biotic stress resistance, boosting nutritional content, etc. In this review, we assess the potential and limitations for (1) identifying specialized metabolic pathways in plants with multi-omics tools and (2) using these enzymes in synthetic biology or crop engineering. The goal of these topics is to highlight areas of research that may need further investment to enhance the successful application of synthetic biology for exploiting the myriad of specialized metabolic pathways.

Achieving convergent synthetic strategies has long been a gold standard in constructing complex molecular skeletons, allowing for the rapid generation of complexity in comparatively streamlined synthetic routes. Traditionally, biocatalysis has not played a prominent role in convergent laboratory synthesis, with the application of biocatalysts in convergent strategies primarily limited to the synthesis of chiral fragments. Although the use of enzymes to enable convergent synthetic approaches is relatively new and emerging, combining the efficiency of convergent transformations with the selectivity achievable through biocatalysis creates new opportunities for efficient synthetic strategies. This Perspective provides an overview of recent developments in biocatalytic strategies for convergent transformations and offers insights into the advantages of these methods compared to their small molecule-based counterparts.

The influence of ten agro-residues, i.e., wheat straw (WS), poplar wood sawdust (PWS), grape pomace (GP), beech wood shavings (BWS), cotton cake (CK), corn cobs (CC), coffee residue (CR), olive pulp (OP), barley and oats straw (BOS) and rice bark (RB), used as alternative substrates for the cultivation of Pleurotus mushrooms, was examined during mycelial growth (colonization phase). Several native and commercial strains of P. ostreatus and P. eryngii were subjected to screening regarding their mycelial growth rates, biomass production and endoglucanase and laccase activities. The experimental results showed that the highest growth rates of P. ostreatus strains were noticed on BWS, BOS, CC and RB substrates, with the time of complete colonization varying between 16 and 36 days, whereas for P. eryngii a high linear growth rate was observed on CC, OP, CR and BOS with the colonization period ranging between 26 and 51 days. The maximum biomass production obtained on the various substrates for both Pleurotus species ranged from 115.32 to 454.42 mg/g d.w. for P. ostreatus and from 108.50 to 422.59 mg/g d.w. for P. eryngii. As for endoglucanase, the highest activities were observed in P. eryngii AMRL 163 cultures on PWS and BOS, i.e., 0.19 and 1.18 U/g d.w., respectively. P. eryngii AMRL 173–6 and P. ostreatus AMRL 150 cultivated on RB and BOS, synthesized significant laccase amounts, i.e., 2172.28 and 1987.25 U/g d.w., respectively. The effect of the substrate components on the growth parameters was considered and discussed. This study showed the industrial potential to convert important low-value agro-residues to fungal biomass and enzymes and eventually to valuable food products.

RESUMEN La butirilcolinesterasa es una enzima que metaboliza relajantes neuromusculares despolarizantes como la succinilcolina, fármaco de elección para procedimientos que requieran parálisis muscular a corto plazo como facilitar la intubación endotraqueal en pacientes sometidos a procedimientos de emergencia. La deficiencia de butirilcolinesterasa se define como la reducción cuantitativa de dicha enzima y su actividad para hidrolizar moléculas, constituyéndose en la principal causa de bloqueo neuromuscular prolongado tras la administración de relajantes neuromusculares como la succinilcolina. Es una condición patológica que puede ser de origen hereditario o adquirido; siendo más común la deficiencia enzimática de origen genético y de carácter autosómico recesivo, la cual se presenta aproximadamente en una de cada 3200 a 5000 personas en todo el mundo. Su manifestación clínica se caracteriza por relajación muscular persistente, la cual puede producir insuficiencia respiratoria aguda. El diagnóstico debe estar orientado a la identificación de sus características clínicas, la cuantificación serológica y el monitoreo neuromuscular. Debido a que no existe cura para esta deficiencia, el manejo debe estar orientado a realizar ventilación mecánica del paciente hasta que el medicamento empleado se metabolice por completo. Este artículo tiene como objetivo realizar una revisión del estado del arte, describiendo su epidemiología, etiología, fisiopatología, manifestaciones clínicas y actualidades en su diagnóstico y tratamiento. ABSTRACT Butyrylcholinesterase is an enzyme that metabolizes depolarizing neuromuscular relaxants, such as succinylcholine, a chosen medication for procedures that require short-term muscular paralysis, to facilitate endotracheal intubation in patients undergoing emergency procedures, for example. Butyrylcholinesterase deficiency can be defined as a quantitative reduction of the enzyme and its activity to hydrolyze molecules, becoming the main cause of prolonged neuromuscular blockade after the administration of neuromuscular relaxants such as succinylcholine. It is a pathological condition that can be of either hereditary or acquired origin; being more common the enzymatic deficiency of genetic origin and of auto-somal recessive character, occurring in approximately one in 3,200 to 5,000 people worldwide. Its clinical manifestation is characterized by persistent muscle relaxation which can lead to acute respiratory failure. The diagnosis must be oriented to the identification of its clinical characteristics, serological quantification, and neuromuscular monitoring. Because a cure does not exist for this deficiency, management should be directed to mechanical ventilation of the patient, until the used drug is fully metabolized. This article aims to review the state of the art, describing its epidemiology, etiology, pathophysiology, clinical manifestations, and updates in its diagnosis and treatment.

A diet low in fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAPs) is a promising therapeutic approach to reduce gastrointestinal symptoms associated with irritable bowel syndrome (IBS). However, a shift toward a more sustainable, healthy diet with higher inclusion of whole-grain cereals (i.e., wheat, rye, barley) and pulses, naturally rich in FODMAPs, poses a severe challenge for susceptible individuals. Dietary restriction of fermentable carbohydrates (commonly called the "low FODMAP diet") has received significant consideration. Hence, the development of functional low FODMAP products is emerging in food science and the food industry. In this review, we evaluate the most promising yet neglected (bio)-technological strategies adopted for modulating the FODMAP contents in complex food systems and the extent of their uptake in the global food market. We extensively investigated the global low FODMAP market, contrasted with the status quo in food science and discussed the key principles and concomitant challenges of targeted FODMAP reduction strategies. Powerful tools are available which are based either on the use of ingredients where FODMAPs have been physically removed (e.g., by membrane filtration) or biotechnologically reduced during the food processing, mediated by added enzymes, microbial enzymes during a fermentation process, and seed endogenous enzymes. However, 10% of the small market of functional products with a low FODMAP claim (total ∼800 products) used any of the targeted FODMAP reduction techniques. The global market is currently dominated by gluten-free products, which are naturally low in FODMAPs and characterized by inferior sensory attributes.

The quest for novel broad spectrum bioactive compounds is needed continuously because of the rapid advent of pathogenic multi drug resistant organisms. Actinomycetes, isolated from unexplored habitats can be a solution of this problem. The motive of this research work was isolation of actinomycetes having potential antimicrobial activities from unexplored regions of Devbag and Tilmati beach. The isolated actinomycetes were screened against pathogenic microbes for antimicrobial activities through cross streak method. Enzyme production activity was checked for these actinomycetes for amylase, protease, cellulase and lipase enzymes. Further antimicrobial activity of ethyl acetate extract of the potent strain KS46 was performed. The strain KS46 was identified with 16S rRNA gene sequencing and secondary structure was analysed. Gas chromatography-Mass spectrometry (GC-MS) profiling was conducted to ascertain the presence of bioactive metabolites in the ethyl acetate extract. The collected samples were pre-treated and 70 actinomycetes were isolated. The Streptomyces sp. strain KS46 showed the best antimicrobial activity in primary screening. Ethyl acetate extract of the strain KS46 revealed antimicrobial activity against S. aureus, B. subtilis, B. cereus, E. faecalis, K. pneumoniae, E. coli, S. flexneri, C. albicans and C. glabrata. The 16S rRNA gene sequencing identified the strain KS46 as Streptomyces levis strain KS46. The GC-MS metabolite profiling of the ethyl acetate extract revealed the availability of 42 compounds including fatty acid esters, fatty acid anhydrides, alkanes, steroids, esters, alcohols, carboxylic ester, etc. having antibacterial, antifungal, antiproliferative, antioxidant activities. This study indicated that Devbag and Tilmati beaches being untapped habitats have enormous diversity of promising antimicrobial metabolite producing actinomycetes. Therefore, further exploration should be carried out to characterize the potential actinomycetes, which can be optimistic candidates for generation of novel antimicrobial drugs.

Dyslipidemia, a major contributor to cardiovascular diseases, is rapidly increasing in Asian countries including Bangladesh. In addition to the cardiovascular system, abnormal lipid levels are also known to cause complications in renal and hepatic systems. The data regarding dyslipidemia and its relationship with liver enzymes are scarce for the Bangladeshi population. Therefore, this study was conducted to estimate the prevalence of dyslipidemia and determine the relationship between lipid profile and liver enzymes in Bangladeshi adults. A total of 405 participants (318 males and 87 females) were enrolled in the study. Serum levels of TG, TC, LDL, HDL and liver enzymes including ALT, AST, GGT and ALP were analyzed using standard methods. Dyslipidemia and liver function tests abnormalities were defined according to the international standard guidelines. The association between elevated lipid profile markers and liver enzyme abnormalities was assessed by logistic regression analysis. Overall, the prevalence of elevated TG, TC, LDL and low HDL were 30.9%, 23.7%, 26.2% and 78.8%, respectively. On the other hand, the prevalence of elevated liver enzymes ALT, AST, GGT and ALP were 18.8%, 21.6%, 12.9% and 21.9%, respectively. Dyslipidemia and liver enzyme abnormalities were higher in diabetic and hypertensive participants than in the healthy participants. About 61% of participants with dyslipidemia had at least one or more elevated liver enzymes. In regression analysis, an independent association was observed between serum GGT and all lipid components. In conclusion, a high prevalence of dyslipidemia and liver enzyme abnormalities were observed among the study participants. Of the four liver enzymes, the serum levels of GGT showed an independent association with all lipid components. Moreover, this study indicates that subjects with dyslipidemia often have a higher chance of having liver diseases than subjects with no dyslipidemia. However, large-scale prospective studies are needed to understand the underlying mechanisms of lipid-induced hepatic dysfunction in the Bangladeshi population.

Superoxide dismutases are important group of antioxidant metallozyme and play important role in ROS homeostasis in salinity stress. The present study reports the biochemical properties of a salt-tolerant Cu, Zn-superoxide from Avicennia marina (Am_SOD). Am_SOD was purified from the leaf and identified by mass-spectrometry. Recombinant Am_SOD cDNA was bacterially expressed as a homodimeric protein. Enzyme kinetics revealed a high substrate affinity and specific activity of Am_SOD as compared to many earlier reported SODs. An electronic transition in 360–400 nm spectra of Am_SOD is indicative of Cu2+-binding. Am_SOD activity was potentially inhibited by diethyldithiocarbamate and H2O2, a characteristic of Cu, Zn-SOD. Am_SOD exhibited conformational and functional stability at high NaCl concentration as well in alkaline pH. Introgression of Am_SOD in E. coli conferred tolerance to oxidative stress under highly saline condition. Am_SOD was moderately thermostable and retained functional activity at ~ 60 °C. In-silico analyses revealed 5 solvent-accessible N-terminal residues of Am_SOD that were less hydrophobic than those at similar positions of non-halophilic SODs. Substituting these 5 residues with non-halophilic counterparts resulted in > 50% reduction in salt-tolerance of Am_SOD. This indicates a cumulative role of these residues in maintaining low surface hydrophobicity of Am_SOD and consequently high salt tolerance. The molecular information on antioxidant activity and salt-tolerance of Am_SOD may have potential application in biotechnology research. To our knowledge, this is the first report on salt-tolerant SOD from mangrove.