Search

Abstract: Bacteriophages are considered as a potential alternative to fight pathogenic bacteria during the antibiotic resistance era. With their high specificity, they are widely used in various applications: medicine, food industry, agriculture, animal farms, biotechnology, diagnosis, etc. Many techniques have been designed by different researchers for phage isolation, purification, and amplification, each of which has strengths and weaknesses. However, all aim at having a reasonably pure phage sample that can be further characterized. Phages can be characterized based on their physiological, morphological or inactivation tests. Microscopy, in particular, opened a wide gate, not only for visualizing phage morphological structure, but also for monitoring biochemistry and behavior. Meanwhile, computational analysis of phage genomes provides more details about phage history, lifestyle, and the potential for toxigenic or lysogenic conversion, which translate to safety in biocontrol and phage therapy applications. This review article summarizes phage application pipelines at different levels, and addresses specific restrictions and knowledge gaps in the field. Recently developed computational approaches, which are used in phage genome analysis, are critically assessed. We hope that this assessment provides researchers with useful insights for the selection of suitable approaches for phage-related research aims and applications.

Diseases with a significant loss of neurons, structurally and functionally are termed as neurodegenerative diseases. Due to the present therapeutic interventions and progressive nature of diseases, a variety of side effects have risen up, thus leading the patients to go for an alternative medication. The role of medicinal plants in such cases has been beneficial because of their exhibition via different cellular and molecular mechanisms. Alleviation in inflammatory responses, suppression of the functionary aspect of pro-inflammatory cytokines like a tumor, improvement in antioxidative properties is among few neuroprotective mechanisms of traditional plants. Variation in transcription and transduction pathways plays a vital role in the preventive measures of plants in such diseases. Neurodegenerative diseases are generally caused by the depletion of proteins, oxidative and inflammatory stress, environmental changes and so on, with aging being the most important cause. Natural compounds can be used in order to treat neurodegenerative diseases Medicinal plants such as Ginseng, Withania somnifera, Bacopa monnieri, Ginkgo biloba, etc. are some of the medicinal plants for the prevention of neurological symptoms. This review deals with the use of different medicinal plants for the prevention of neurodegenerative diseases.

Background Population growth, climate change, and food shortage have led to increased demand for alternate sources of lipids to meet our food and energy needs. Oleaginous microorganisms such as yeasts, bacteria, microalgae, fungi, and thraustochytrids are good resources for producing lipids. Scope and approach Many factors affect the amount and composition of lipid accumulation in microorganisms, such as genetic makeup and environmental conditions during growth, such as carbon and nitrogen sources, pH, temperature, and exposure to natural light in growing media. Food biotechnology advances such as improvements in fermentation technology, genetic and metabolic engineering of oleaginous microorganisms, and the use of low-cost carbon sources, such as food and agricultural waste materials, make lipid production using microbial sources more feasible. Key findings and conclusions Microbial lipid production has some unique benefits for the production of specialty fats and oils, including cocoa butter equivalents, polyunsaturated fatty acid oils and biosurfactants. Moreover, microbial oil production has advantages over plant and marine oils, including lower cultivation costs and the ability to engineer oil structure and thus functionality using genetic modification. Microbial oils production allows for the production of more sustainable and safer oils, without pesticides, herbicides or heavy metals, year-round with a small footprint and without the need for land.

Larvae of Tenebrio molitor (yellow mealworm; YM) are an alternative to conventional animal protein as food and feed thanks to their high nutritional value and low environmental impact for production. Since some studies have reported high water activity (aw) with different microbial contamination levels, blanching and drying can be employed to reduce microbial load, enzymatic activity, moisture content, and water activity, ensuring safety and preserving YM for longer periods. This study applied rinsing or blanching followed by vacuum- or multiflash drying, aiming to preserve YM by reducing aw and the microbial load, enabling its use as food and feed. The processing impact on YM colour was evaluated, and they were also subjected to an Escherichia coli contaminated wheat bran to perform a challenge test to evaluate the effectiveness of the decontamination. Drying curves were determined for each situation. Multiflash drying procedure was faster than vacuum drying for reducing YM moisture. Additionally, blanched samples required a shorter period to reach the targeted water activity (0.2-0.3) than rinsed samples. Blanching plus vacuum drying reduced more than 5 log E. coli load in YM in the challenge test. Vacuum drying preserved YM brightness the most. In conclusion, this study showed that drying could provide shelf-stable YM. Besides, only 15 s of blanching before drying gives better brightness and leads to a greater microbial reduction after a relatively short drying time.

In August 2016, tomato plants grown during a hot, wet summer with heavy soil flooding, displaying symptoms of wilting, dead plant, root rot with crown and stem rot, at Beni Suef and Fayoum governorates were examined. A number of 16 fungal isolates were isolated from tomato plants displaying the above symptoms. These isolates were classified as belonging to six species, namely: Alternaria solani, Chaetomium globosum, Fusarium solani, Fusarium oxysporum, Pythium spp. and Rhizoctonia solani. Isolates of Pythium spp. were prevalent and were found to be more pathogenic than the other fungal isolates. This species causes damping-off, root rot, sudden death, stem rot and fruit rot. The pathogen was identified as Pythium aphanidermatum based on morphological, cultural, and molecular characteristics. Biogenic silver nanoparticles (AgNPs) were produced using the F. oxysporum strain and characterized by transmission electron microscopy (TEM). The size of these spherical particles ranged from 10 to 30 nm. In vitro, biogenic AgNPs showed antifungal activity against P. aphanidermatum. In greenhouse and field experiments, AgNPs treatment significantly reduced the incidence of dead tomato plants due to root rot caused by P. aphanidermatum compared to the control. All of the investigated treatments were effective and the treatment of root dipping plus soil drenching was the most effective. To the best of our knowledge, this study describes P. aphanidermatum on tomato in Egypt for the first time. Also, biogenic AgNPs could be used for controlling root rot disease caused by this pathogen.

Ergothioneine is a naturally occurring antioxidant that has shown potential in ameliorating neurodegenerative and cardiovascular diseases. In this study, we investigated the potential of the Crabtree-negative, oleaginous yeast Yarrowia lipolytica as an alternative host for ergothioneine production. We expressed the biosynthetic enzymes EGT1 from Neurospora crassa and EGT2 from Claviceps purpurea to obtain 158 mg/L of ergothioneine in small-scale cultivation, with an additional copy of each gene improving the titer to 205 mg/L. The effect of phosphate limitation on ergothioneine production was studied, and finally, a phosphate-limited fed-batch fermentation in 1 L bioreactors yielded 1.63 ± 0.04 g/L ergothioneine in 220 hours, corresponding to an overall volumetric productivity of 7.41 mg L-1 h-1 , showing that Yarrowia lipolytica is a promising host for ergothioneine production.

BACKGROUND: The major component of lemongrass by weight is dietary fibre (DF), but no literature has been reported on its DF components and fermentation products when ingested. OBJECTIVE: This study analysed DF components of lemongrass, investigated the potential of the major product from total DF (TDF) fermentation to inhibit α-amylase and HMG-CoA reductase, key enzymes of diabetes mellitus and hypercholesterolemia, respectively, and determined the serum glucose- and cholesterol-lowering potential of TDF in an animal model. METHODS: Lemongrass DF components were analysed, TDF was fermented in vitro; the major fermentation product was isolated for enzyme inhibitory assays; and postprandial blood glucose- and cholesterol-lowering potential of TDF was determined in Sprague-Dawley rats. RESULTS: TDF in lemongrass was 65.7 g/100g and soluble DF, 2.8 g/100 g. Significant amount of propionate (10.9 mM/g TDF) was produced after TDF fermentation; propionate inhibited 20.4% α-amylase activity, and 13.1 % HMG-CoA reductase activity in vitro. TDF further exhibited antihyperglycemic and cholesterol-lowering potential in an animal model. CONCLUSIONS: DF from lemongrass was shown to reduce hyperglycemia and hypercholesterolemia in an animal model, through mild inhibition of α-amylase and HMG-CoA reductase. Thus, lemongrass DF may have a significant role in mitigating the risk of type 2 diabetes mellitus and hypercholesterolemia.

Background Food fermentation has been practiced since prehistoric times and developed over the years with modification in substrates, processes and technologies. It is carried out by microbial cultures with techniques such as back slopping and enrichment, enhancing the organoleptic property, nutrient availability and storage life of food and in many cases contributing beneficial microbes to the diet of the consumer. Scope and approach The microorganisms involved in fermentation greatly vary according to the food product. One of the major groups of microbes used in traditional and industrial fermentation of cereals includes the lactic acid bacteria, many of which are known to have probiotic characteristics followed by yeast and mould. While dairy fermented foods have received attention as a source of probiotic microbes, those in non-dairy foods especially millets and cereal mixtures have not been documented. Traditional fermented foods of animal origin as well as other plant foods such as pulse, fruits and vegetables are also discussed. The review examines the presumptive probiotic bacteria found in non-dairy fermented foods around the globe. Key findings and conclusions This review is an attempt to collate the scientific progress with respect to probiotics in millet fermented foods, particularly of Africa and Asia. India has the largest documented number of fermented foods followed by Korea especially made with cereals. These fermented foods containing presumptive probiotic microorganisms may serve as probiotic supplements that are affordable as opposed to expensive probiotic formulations.

This study was conducted to compare the correlations between free radical antioxidant activity and anti-wrinkling activity. Areca nut was extracted using various concentrations of ethanol (0, 40, 80, and 100%) and methanol (0, 40, 80, and 100%) at 100 °C for 90 min. The ethanolic and methanolic extracts of Areca nut were evaluated for total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity, including total antioxidant capacity (TAC), FRAP, DPPH, ABTS, and superoxide anion radical scavenging activity (SRSA), and anti-wrinkling activity (collagenase inhibition and elastase inhibition). In addition, the antioxidant activities were correlated with collagenase inhibition. The antioxidant activities of the extracts exhibited little correlation (R=0.008-0.0760) with TPC, TFC, and TAC; however, significant correlation (R=0.825-0.891) was shown with collagenase inhibition. The 40% and 80% EtOH extracts showed the highest levels of antioxidant activities, and also had the highest phenolic and flavonoid contents. The antioxidant activities of the 80% EtOH extracts was greater than that of other extracts. These results indicate that the collagenase inhibition effects of Areca nut extracts may be due to free radical antioxidant activities, and the optimal extraction condition was 80% EtOH.

Vine shoots (Vitis vinifera L.) constitute an abundant lignocellulosic source which is frequently underutilised. Alkaline and acidic pretreatments (with and without washing steps) were compared and optimised to release fermentable sugars from vine shoots. An acidic pretreatment using 1.72% H2SO4 at 134 °C for 17 min (with 10% w/w solid biomass), followed by an enzymatic hydrolysis, offered the most cost-effective results, releasing 40.21 g/L sugars. Three thermotolerant strains, namely, Bacillus coagulans DSM 2314, Geobacillus stearothermophilus DSM 2313, and G. stearothermophilus DSM 494, were assessed to produce lactic acid from vine-shoot hydrolysates under aerobic and non-sterile conditions, without the need of detoxification steps. In addition, wine lees were satisfactorily employed as nitrogen sources for the fermentation, providing similar results to yeast extract and being the only nutrient added to vine-shoot hydrolysates. Under optimal conditions, B. coagulans DSM 2314 produced 29.21 ± 0.23 g/L lactic acid in 24 h, with a sugar consumption of 98.74 ± 0.07% and a yield of 96.38 ± 0.76%, when supplemented with red wine lees. The purity of the isomer L( +) reached 97.59 ± 1.35% of the total lactic acid produced. Although G. stearothermophilus was able to transform the hexoses from vine-shoot hydrolysates into lactic acid, it proved to be inefficient for metabolising pentoses, thus obtaining lower lactic acid values (16–18 g/L).

Microalgae can produce industrially relevantmetabolites using atmospheric CO2 and sunlight as carbon and energy sources, respectively. Developing molecular tools for high-throughputgenome engineering could accelerate the generation oftailored strains with improved traits. To this end, we developed a genome editing strategy based on Cas12a ribonucleoprotein (RNPs) and homology-directed repair (HDR) to generate scarlessand markerless mutants of the microalga Nannochloropsis oceanica. We also developed an episomal plasmid-based Cas12a system forefficiently introducing indels at the target site. Additionally, weexploited the ability of Cas12a to process an associated CRISPRarray to perform multiplexed genome engineering. We efficientlytargeted three sites in the host genome in a singletransformation,thereby making a major step toward high-throughput genome engineering in microalgae. Furthermore, a CRISPR interference(CRISPRi) tool based on Cas9 and Cas12a was developed for effective downregulation of target genes. We observed up to 85%reduction in the transcript levels upon performing CRISPRi with dCas9 in N. oceanica. Overall, these developments substantiallyaccelerate genome engineering efforts in N. oceanica and potentially provide a general toolbox for improving othermicroalgal strains.

Introduction. Grape pomace is the most important by-product of winemaking that can be used as an additional raw material. There is a need for an optimal storage technology so that pomace can be further processed to obtain new types of products. We aimed to study the effect of grape pomace treatment on its microflora. Study objects and methods. We identified and quantified microflora on the fresh and one-month-stored pomace samples from white and red grape varieties. The samples were exposed to conventional drying at 60–65°C, infrared drying at 60–65°C, as well as sulfitation with sulfur dioxide and sodium metabisulfite. Results and discussion. The pomace microflora can be considered a microbial community. Almost all the samples stored for one month in an open area contained Saccharomyces cerevisiae yeasts, higher concentrations of filmy yeasts of the Candida, Pichia, Hansenula, Hanseniaspora/Kloeckera, and Torulaspora genera, as well as conidia of Mucor, Aspergillus niger, and Penicillium molds. Prevalent bacteria included acetic acid (mainly Acetobacter aceti) and lactic acid (Lactobacillus plantarum, Pediococcus, Leuconostoc) bacteria. These microorganisms significantly changed concentrations of volatile and non-volatile components, decreasing total polysaccharides, phenolic compounds, and anthocyanins 1.7–1.9, 3.7–4.0, and 4.0–4.5 times, respectively. The contents of micromycetes and bacteria in the one-month-stored samples were significantly higher than in the fresh pomace. Predrying and sulfitation decreased bacterial contamination, but to a lesser extent compared to micromycetes. Conclusion. Long-term storage spoiled pomace, leading to significant changes in its chemical composition. Sulfitation reduced microorganism growth during storage, but did not provide long-term preservation (over a month), while pre-drying at 60–65°C promoted longer storage.

While nepetalactone, the active ingredient in catnip, is a potent insect repellent, its low in planta accumulation limits its commercial viability as an alternative repellent. Here we describe a platform for de novo nepetalactone production in Saccharomyces cerevisiae, enabling sustainable and scalable production. Nepetalactone production required introduction of eight exogenous genes including the cytochrome P450 geraniol-8-hydroxylase, which represented the bottleneck of the heterologous pathway. Combinatorial assessment of geraniol-8-hydroxylase and cytochrome P450 reductase variants, as well as copy-number variations were used to overcome this bottleneck. We found that several reductases improved hydroxylation activity, with a higher geraniol-8-hydroxylase ratio further increasing 8-hydroxygeraniol titers. Another roadblock was the accumulation of an unwanted metabolite that implied inefficient channeling of carbon through the pathway. With the native yeast old yellow enzymes previously shown to use monoterpene intermediates as substrates, both homologs were deleted. These deletions increased 8-hydroxygeraniol yield, resulting in a final de novo accumulation of 3.10 mg/L/OD600 of nepetalactone from simple sugar in microtiter plates. Our pathway optimization will aid in the development of high yielding monoterpene S. cerevisiae strains.

Introduction. A disadvantage of the ancestral method (la méthode ancestrale), which is widely used in the production of sparkling wine, is that it is difficult to control fermentation. We aimed to identify the optimal yeast race for obtaining high-quality young sparkling wines with varietal aroma without yeast tones. Study objects and methods. Our study objects were base and young sparkling wines from Cabernet-Sauvignon prepared on various yeast races. Organic acids, sugars, and ethanol contents were determined by high performance liquid chromatography. Phenolic and coloring substances were measured by colorimetric method. Foaming properties were determined by air barbotage of a wine sample in a measuring cylinder; sparkling properties, by measuring the CO2 desorption rate; CO2 content, by volumetric method; viscosity, with a viscometer. Sensory evaluation was carried out according to standard methods. Results and discussion. The wines produced on the Odesskiy Chernyi-SD13 yeast race received the highest tasting scores of 7.82 and 9.05 points for base wine and young sparkling wines, respectively. They contained larger amounts of phenolic substances (1103 mg/dm3) and coloring agents (275 mg/dm3) and had higher color intensity (1.614). The panelists rated them highly on their complex varietal aroma and harmonious, velvety flavor, as well as their foaming and sparkling properties. This yeast race ensured intensive fermentation of sugars and a great amount of bound CO2 (up to 24.93%). Conclusion. The Odesskiy Chernyi-SD13 yeast race is optimal for making base and young sparkling wines by the bottle method. This technology can be used to produce high-quality sparkling wines in the crop year by large and small enterprises.

Fungal pre-treatment and fermentation could improve nutritional value of wet corn distillers’ grains and solubles (WDGS), soybean hull (SH), and their blend in swine and poultry diets, which can potentially increase revenues for corn-ethanol and soybean processing industries while reducing feed cost in swine and poultry production systems. This study evaluated the effectiveness of pre-treatment of SH, WDGS, and their mixture by Trichoderma reesei and fermentation by Aspergillus oryzae to improve the nutritional profile and digestibility of these ingredients. T. reesei produced cellulase and xylanase, resulting in structural carbohydrates reduction by 69.2% while concentrating amino acids from 6.8% to 11% in SH. Fermentation with A. oryzae degraded phytate by over 50% and improved in vitro digestibility of amino acids in WDGS and its mixture with SH. Fermenting T. reesei pre-treated SH by A. oryzae showed higher in vitro dry matter digestibility than non-fermented substrate. Moreover, the proportion of key amino acids (arginine, threonine, methionine, and lysine) in both T. reesei and A. oryzae fermented substrates were significantly improved. The results demonstrated the feasibility of combining T. reesei and A. oryzae in improving feeding value of WDGS and SH for potential use as feed ingredients for monogastric animals.

In recent years, attention to m icrobial dehalogenase has continually increased due to its potential application, both in bioremediation and in the biosynthesis of fine chemicals. Many microbial recombinant strains carrying dehalogenase gene have been developed, particularly to increase the dehalogenase production and its quality. In this study, we aimed to find the optimum condition for the production of active haloacid dehalogenase by E. coli BL21 (DE3) harboring recombinant plasmid pET- bcfd1 that carried haloacid dehalogenase gene from Bacillus cereus IndB1 local strain. This would be examined by assessing the ability of whole cell life culture to degrade monochloroacetic acid (MCA) and quantifying the chloride ion released into the medium. Several variables were evaluated to find this optimal condition. We found that the best condition for MCA biodegradation using this recombinant clone was at 0.2 mM MCA, 10 μM of isopropyl β -D-1-thiogalactopyranoside (IPTG), 6 hours of pre-induction incubation at 37 o C with shaking, 2 hours IPTG induction at 30 o C with shaking, at pH 7 in Luria Bertani (LB) liquid medium without NaCl, which produced about 0.056 mM chloride ions. Inducer concentration, pre-induction incubation time and temperature, as well as induction time and temperature were apparent to be associated with the expression of the protein, while the MCA concentration and the pH of the medium influenced the ability of the recombinant E. coli BL21 (DE3)/pET- bcfd1 to grow in toxic environment. Our findings laid the foundation for exploration of dehalogenases from local Bacillus strains through genetic engineering for MCA biodegradation

Made available in DSpace on 2022-04-28T19:46:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-01-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Academy of Finland We investigated whether highly available organic residues in Brazil can be used as substrates for the production of the oyster mushroom Pleurotus ostreatus, instead of the conventional cultivation using the eucalyptus sawdust substrate. We assessed the mushroom yield on 13 substrates, of which 12 were formulated with different concentrations of organic residues and one with pure eucalyptus sawdust, and verified whether the raw material used in the substrate formula and the concentration of such alternative residues influenced their biological efficiencies. Substrates containing eucalyptus bark resulted in higher mushroom yield than those containing eucalyptus sawdust, which generally resulted in similar mushroom yield to the remaining formulas. Moreover, the raw material and the concentration of each residue affected the biological efficiency of the substrates. We show that the conventional substrate for P. ostreatus can be replaced by substrates easily accessible to producers without loss in productivity. Furthermore, that the concentration of these mixtures affects the mushroom productivity and should be considered when formulating the growth medium. Graduate Program in Agriculture – Energy in Agriculture School of Agricultural Sciences São Paulo State University (UNESP) Department of Microbiology University of Helsinki Graduate Program in Biological Sciences (Botany) Institute of Biosciences São Paulo State University (UNESP) Department of Biodiversity Laboratory of Phenology Institute of Biosciences São Paulo State University (UNESP) Graduate Program in Agriculture – Energy in Agriculture School of Agricultural Sciences São Paulo State University (UNESP) Graduate Program in Biological Sciences (Botany) Institute of Biosciences São Paulo State University (UNESP) Department of Biodiversity Laboratory of Phenology Institute of Biosciences São Paulo State University (UNESP) CAPES: 001 Academy of Finland: 308284

This study aimed to investigate the effect of repeated batch fermentation of cocultured Saccharomyces bayanus Y4 and Torulaspora delbrueckii Y7 on the quality of greengage wine. The acidity, as well as citric acid and L-malic acid content, decreased significantly with the batches. The citric acid content of the single culture in each batch was 58.59, 26.27, and 8.45 g/L, respectively. Additionally, the ester content increased with the batches. The respective values were 1627.93, 2249.46, and 2491.17 μg/L for the prophase of coculture. However, volatile content in the fresh wine of Y4 yeast in each batch was high. Ketone and terpene contents in the first batch of coculture significantly increased from 31.64 to 95.60 μg/L and 58.58–95.13 μg/L, respectively. Furthermore, a significant difference was observed between the microbial communities fermented using single culture and coculture. The change in fungal community diversity for coculture pattern was higher than that in the mash-inoculated single strain, which decreased the abundance of dominant fungi. Pichia, Stropharia, Suillus, Malassezia, Trichoderma, Faecalibacterium, and Butyricicoccus were associated with esters, terpenes, phenylethyl alcohol, and β-ionone. Our study provides a new avenue to improve the flavour and quality of wine brewed using a characteristic fruit.

Microbial enzymes are crucial for material biotransformation during the composting process. In this study, we investigated the effects of adding bamboo charcoal (BC) (i.e., at 5%, 10%, and 20% corresponding to BC5, BC10, and BC20, respectively) on the enzyme activity levels during chicken manure composting. The results showed that BC10 could increase the cellulose and urease activities by 56% and 96%, respectively. The bacterial community structure in BC10 differed from those in the other treatments, and Luteivirga, Lactobacillus, Paenalcaligenes, Ulvibacter, Bacillus, Facklamia, Pelagibacterium, Sporosarcina, Cellvibrio, and Corynebacterium had the most important roles in composting. Compared with other treatments, BC10 significantly enhanced the average rates of degradation of carbohydrates (D-xylose (40%) and α-D-lactose (44%)) and amino acids (L-arginine (16%), L-asparagine (14%), and L-threonine (52%)). We also explored the associations among the bacterial community and their metabolic functions with the changes in the activities of enzymes. Network analysis demonstrated that BC10 altered the co-occurrence patterns of the bacterial communities, where Ulvibacter and class Bacilli were the keystone bacterial taxa with high capacities for degrading carbon source, and they were related to increases in the activities of cellulase and urease, respectively. The results obtained in this study may help to further enhance the efficiency of composting.

Background: Cancer is a non-communicable disease with increasing incidence and mortality rates worldwide, including Thailand. Its apparent lack of effective treatments is posing challenging public health issues. Introduction: Encouraging research results indicating probable anti-cancer properties of the Delonix regia flower Extract (DRE) have prompted us to evaluate the feasibility of developing a type of product for future cancer prevention or treatment. Methods and Results: In the present report, using High-Performance Liquid Chromatography (HPLC), we demonstrate in the DRE the presence of high concentrations of three identifiable flavonoids, namely rutin 4.15±0.30% w/w, isoquercitrin 3.04±0.02 %w/w, and myricetin 2.61±0.01% w/w, respectively, while the IC50 of DPPH and ABTS assay antioxidation activity was 66.88±6.30 μg/ml and 53.65±7.24 μg/ml, respectively. Discussion: Our cancer cell line studies using the MTT assay demonstrated DREs potent and dosedependent inhibition of murine leukemia cell line (P-388: 35.28±4.07% of cell viability remaining), as well as of human breast adenocarcinoma (MCF-7), human cervical carcinoma (HeLa), human oral cavity carcinoma (KB), and human colon carcinoma (HT-29) cell lines in that order of magnitude. Conclusion: Three identifiable flavonoids (rutin, isoquercitrin and myricetin) with high antioxidation activity and potent and dose-dependent inhibition of murine leukemia cell line and five other cancer cell lines were documented in the DRE. The extract’s lack of cytotoxicity in 3 normal cell lines is a rare advantage not usually seen in current antineoplastic agents. Yet another challenge of the DRE was its low dissolution rate and long-term storage stability, issues to be resolved before a future product can be formulated.

This study aimed to evaluate the effects of by-product molasses and waste orange peels (WOP) as the nutrient medium on lipase production with Rhizopus arrhizus NRRL 2286 and investigate the inductive effect of waste cooking oil (WCO) on activity. The results showed that the highest specific growth rate and the maximum biomass concentration were obtained 0.424 h−1 and 3.83 g dry weight/L for 70 g/L WOP, respectively. Additionally, the highest intracellular and extracellular lipase activities were attained in 50 g/L WOP as 400 U/g and 13.2 U/mL, correspondingly. Next, the inductive effects of WCO and olive oil were tested in lipase production. Intracellular lipase activity enhanced and reached a maximum of 520 U/g in the presence of 10 g/L WCO with an increase of 30% compared to the one without WCO. However, both inducers caused a diminish in extracellular lipase hydrolysis activity. Overall, this study reveals that the lipase production of R. arrhizus can be accomplished using WOP and WCO with the optimum conditions.

Amylases have been widely applied for starch saccharification in the food, paper, pharmaceutical, and textile industries. In the present study, α-amylase DL-3-4−1 from B. amyloliquefaciens was successfully expressed in B. subtilis. The enzymatic activity exhibited high stability in a pH range from 4.0 to 11.0 and moderate thermostability. Furthermore, excellent stability was observed when incubated with metal ions, detergents, inhibitors, and organic solvents. Resistance against ethanol, urea, and guanidine−HCl was also observed. More importantly, a strategy to enhance the extracellular expression of α-amylase DL-3-4−1 was developed using 173 Sec-type signal peptides (SPs) as a library based on a fast and sequence-independent method. Fourteen SPs were identified with significantly increased yields of DL-3-4−1 via a high-throughput assay. Specifically, the enzymatic activity of the α-amylase-producing strain containing the signal peptide SP5D10 (YomL) was 3.74-fold higher than that of the strain with the wild type signal peptide (87.30 U/OD). In brief, the maximum specific activity of an α-amylase-producing strain containing the YomL signal peptide was 326.45 U/OD. This is the first report of an α-amylase that exhibits ethanol, urea, and guanidine−HCl tolerance. Hence, this α-amylase represents a promising candidate for future use in industry.

Biological pretreatment of lignocellulose by white-rot fungi is an environmentally friendly alternative to chemical and physical approaches to enhance enzymatic saccharification. However, inefficient lignin degradation and substantial cellulose consumption during fungal pretreatment can cause low sugar yields. In this study, the combined action of CuSO4 and MnSO4 effectively improved the degree and selectivity of delignification during solid-state fungal pretreatment of poplar wood by Phanerochaete chrysosporium. Compared to pretreatment without this supplementation, a 1.9 fold higher lignin degradation and 2.4 times higher delignification selectivity value were obtained due to the increased and prolonged manganese peroxidase activity. Enzymatic saccharification of supplemented pretreated wood resulted in a 2.9 times higher glucose yield compared to the non-supplemented system. This study has demonstrated that the combined application of Mn2+ and Cu2+ can significantly improve the fungal pretreatment process and that the beneficial effect of Cu2+ on delignification is not restricted to laccase-producing fungi.

Ethyl carbamate (EC), widely found in alcoholic beverages, has been revealed to be a probable carcinogen in humans. Urethanase (EC 3.5.1.75) is an effective enzyme for the degradation of EC; however, the previously identified urethanases exhibited insufficient acid and alcohol resistance. In this study, an enantioselective amidase (AmdA) screened from Agrobacterium tumefaciens d3 exhibited urethanase activity with excellent alcohol resistance. AmdA was first overexpressed in Escherichia coli; however, the recombinant protein was primarily located in inclusion bodies, and thus, co-expression of molecular chaperones was used. The activity of AmdA increased 3.1 fold to 307 U/L, and the specific activity of urethanase with C-terminal His-tags reached 0.62 U/mg after purification through a Ni–NTA column. Subsequently, the enzymatic properties and kinetic constants of AmdA were investigated. The optimum temperature for AmdA was 55 °C, it showed the highest activity at pH 7.5, and the Km was 0.964 mM. Moreover, after 1 h of heat treatment at 37 °C in a 5–20% (v/v) ethanol solution, the residual urethanase activity was higher than 91%, considerably more than that reported thus far.

GLOBOCAN estimates in 2020 on the basis of occurrence, mortality, and development for 36 distinct types of tumor cells worldwide. Chemotherapy is the most preferred treatment for carcinoma, and also for hormonal, radiation, autoimmune, and genetic disorders. We propose a straightforward and cost-effective way to overcome the myriad barriers to the therapeutic potential of irinotecan (SN-38), including the rebuilding of medicines, targeting of supramolecular nanoassemblies, and therapeutic treatments. The drug SN-38 was capable by covalent combination of lipophilic linoleic acid by the end of carboxyl group action to provide an impulsive nanoassembly to extremely stable nanomaterials (NMs). The morphology of the recently synthesized SN-38-NM was tested by electron microscopy. The anticarcinoma properties of both HeLa and CaSki (cervical carcinoma lines) of SN-38 and SN-38-NMs were evaluated after successful synthesis. Further research on mechanistic cell death pathways showed that proliferation of cervical carcinoma cell lines was correlated with cells subjected to apoptosis, including AO/EB staining, nuclear staining, and flow cytometric studies. SN-38-NMs have higher biocompatibility in conjunction with free SN-38. This report describes SN-38-NMs as a positive and secure treatment device for cervical carcinoma, which requires further clinical assessment.

The etiological agent of melioidosis, Burkholderia pseudomallei, utilises a type III secretion system cluster 3 (T3SS3) to deliver proteins termed type III effectors (T3SEs) into the host cytoplasm in order to establish an intracellular lifecycle in phagocytic and non-phagocytic cells, thus playing an important role in pathogenesis. To gain insight into possible functional roles for BapA, a putative T3SE with unknown function, in the intracellular lifecycle of B.pseudomallei, bapA gene knockout mutant was constructed. The effect of the knockout on virulence to the otherwise isogenic parental strain, K96243, was studied by cellular infection assays and Caenorhabditis elegans killing assay. The attachment and subsequent entry into A549 cells was significantly (P < 0.05) attenuated in the Delta bapA compared to K96243. However, intracellular replication was not affected. Furthermore, the cell-to-cell spreading capacity of Delta bapA was impaired although the mutant exhibited no evident defect in its actin tail formation. Additionally, phagocytosis and intracellular replication rates of Delta bapA in U937 macrophage cells were significantly reduced relative to K96243 without phagosomal escape being affected. Based on these observations, we conclude that the BapA T3SE could play an important role in B.pseudomallei intracellular lifecycle, especially, in the early stages of attachment and entry into the host cell.

In human milk fat, palmitic acid (16:0) is esterified to the middle (sn-2 or β) position on the glycerol backbone and oleic acid (18:1) predominantly to the outer positions, giving the triacylglycerol (TG) a distinctive stereoisomeric structure that is believed to assist nutrient absorption in the infant gut. However, the fat used in most infant formulas is derived from plants, which preferentially esterify 16:0 to the outer positions. We have previously showed that the metabolism of the model oilseed Arabidopsis thaliana can be engineered to incorporate 16:0 into the middle position of TG. However, the fatty acyl composition of Arabidopsis seed TG does not mimic human milk, which is rich in both 16:0 and 18:1 and is defined by the high abundance of the TG molecular species 1,3-olein-2-palmitin (OPO). Here we have constructed an Arabidopsis fatty acid biosynthesis 1-1 fatty acid desaturase 2 fatty acid elongase 1 mutant with around 20% 16:0 and 70% 18:1 in its seeds and we have engineered it to esterify more than 80% of the 16:0 to the middle position of TG, using heterologous expression of the human lysophosphatidic acid acyltransferase isoform AGPAT1, combined with suppression of LYSOPHOSPHATIDIC ACID ACYLTRANSFERASE 2 and PHOSPHATIDYLCHOLINE:DIACYLGLYCEROL CHOLINEPHOSPHOTRANSFERASE. Our data show that oilseeds can be engineered to produce TG that is rich in OPO, which is a structured fat ingredient used in infant formulas.

The demand for wild tambaqui has led to overfishing of their natural stocks, and has thus increased the demand for farmed fish. This has raised a question regarding the nutritional value of these groups of individuals; to see if they differ when they grow in wild and farmed environments. The meat of 28 tambaqui was evaluated by measuring the moisture, ash and total proteins among the fish from the two different environments in the states of Amazonas and Rondônia, Brazil. The averages of the centesimal composition showed significant differences, between environments and states. The moisture values of wild fish between the states did not differ, but were higher than those farmed in either of the states. The ash value also showed differences between groups by environment and between states. However, it was found that wild tambaqui from both states exhibited the best levels of protein and did not show statistical dissimilarities. The group farmed in Rondônia was also similar to the wild ones caught in that state. As for farmed fish, the averages between states were different, with the lowest value attributed to fish from Amazonas. The data show that wild tambaqui had a higher nutritional value than farmed fish.

Application of plant-mediated Microbial fuel cell (PMFC) for bioelectricity production with food crop yield has not yet been appropriately investigated. This study aims to reveal the ability of two essential food plants: Amaranthus viridis (P1) and Triticum aestivum (P2), for sustainable bioenergy production and food harvest for 180 days consecutively. The study showed that the highest power density obtained from P2-PMFC was higher than the P1-PMFC, and the organic-rich nutrient called bioslurry in PMFCs shows twofold intensification in the electrochemical reaction. The main role of bioslurry (brown gold) as a magical organic-rich nutrient increases the plant growth and the activity of electrogenic bacteria in PMFCs. Thus the current density of P1 (114.52 ± 20.05 mA/m2) and P2 (185.23 ± 15.10 mA/m2) without bioslurry was increased with a maximum current density of 185.23 ± 15.10 mA/m2 (P1) and 291.23 ± 7.50 mA/m2 (P2) with bioslurry. The high content screening technique and scanning electron micrograph confirm the biofilm formation on the rhizosphere-anode region of both the plants. Culturable rhizosphere-anode region bacterial species were isolated in P1 and P2 PMFCs, and their electrogenic activity was confirmed using the single-chambered Microbial fuel cells (MFCs) through polarization test. The constructed phylogenetic tree for the 31 isolates concludes the diversity of all bacterial species, of which the phylum Firmicutes dominates in both PMFCs. The individual polarization curve efficiency of the isolates ranges from 7.62 to 194.45 mW/m2, and among the 31 isolates, Hydrogenophaga sp. JBP18, Pseudomonas aeruginosa JBP1, Ralstonia sp. JBP24, Ochrobactrum intermedium strain JBP22 and Bacillus safensis JBP6 were selected as potential electrogenic bacteria. Accordingly, this study might lead to the development of MFC technology for sustainable electricity production using food crop plants and determining more rhizosphere-anode electrogenic bacterial species in the future.

In Cognac, the musts are rich in grape solids and fermentations are usually run with turbidities ranging between 500 and 1500 NTU (nephelometric turbidity unit). These conditions, considered favourable for generating the desired organoleptic profiles of the final Eaux-de-vies, are unusual in winemaking, and, consequently, their impact on yeast metabolism is poorly understood. This study aims to better describe and understand the synthesis of fermentative aromas in such lipid-excess conditions, while integrating the effect of two other very important parameters: the initial concentration of assimilable nitrogen and the temperature of fermentation. To reach this objective, a Box-Behnken design was implemented to describe and model the simple effects of these factors as well as their interactions. Although the lipid concentration was very high, impacts on the production of fermentative aromas were observed. Indeed, high lipid levels promoted the synthesis of higher alcohols. Observing this effect was surprising because there is no metabolic connection between the anabolic pathways of production of these alcohols and the lipid pathway. This effect may be partly explained by impairment in the activity of alcohol acetyl transferases in the presence of lipids, which catalyse the conversion of higher alcohols into the corresponding esters. Therefore, in this study, the negative impact of turbidity was very significant on acetate esters related to the production of acetyl-CoA, which was the main molecule disturbed by the strong presence of lipids. Finally, and more surprisingly, lipid intake did not impact the synthesis of ethyl esters, which depended on the concentration of exogenous lipids. KEY POINTS: • Innovative work on the fermentation of white wine musts with very high lipid contents. • Precise fermentation management and monitoring in Cognac-making conditions. • Experimental design to study the impact of lipids, assimilable nitrogen and temperature on fermentative aroma synthesis.

Essential oils (EO) are volatile compounds with complex chemical compositions that are derived from the secondary metabolism of aromatic herbs. There are several applications of EO in the industrial and medical sectors. Basil (Ocimum sp.) is one of the most important EO-producing aromatic herbs. In this study, EO content (EOC) and linalool content (LC) in basil irrigated with reclaimed water were investigated. Plant development parameters, nutrient absorption and crude protein (CP) content were also evaluated. The experiment was conducted in a greenhouse, with five different treatments and five repetitions each. Compared with treatments only using water, the results of those with reclaimed water irrigation showed higher nutrient absorption, CP contents (in their foliar tissue), length of plant branches, foliar biomass weights, and EOC and LC. EOC reached up to 0.58% of the dry biomass and LC was up to 5.84% of EO mass. In practice, it is estimated that around 5.8 kg of EO and 0.34 kg of linalool can be obtained from one ton of dry basil biomass. HIGHLIGHTS Higher essential oil and linalool contents were obtained in plants irrigated with higher percentage of reclaimed water used.; Reclaimed water use resulted in the highest length of branches and biomass production of dry leaves, up to 30 cm and 28 g per plant, respectively.; Crude protein content in leaves increased proportionally to the percentage of reclaimed water used.; Approximately 5.8 kg of EO and 0.34 kg of linalool can be obtained per ton of basil dry mass.

Monascus pigments are the important natural additives in food industrial production. To obtain more economic pigments production processes, the present study was performed to evaluate the feasibility of using pomace resource as substrate for pigments production. Petri dish fermentation was designed to seek the optimal process parameters, and the value of red, yellow and total pigments per dry fermented substrate could achieve 654.6, 1268.1 and 1922.7 OD units/g, respectively. Shallow tray fermentation experiments were used for investigating the potential industrial production of pigments using potato pomace as sole carbon. The final total pigments of 200 g and 1000 g shallow tray experiments could reach 1886.9 and 1737.4 OD units/g. The results in this work indicating that potato pomace could be an efficient and low cost substrate for the production of Monascus pigments, and will supply a valuable reference for the comprehensive utilization of potato resources and seeking the economical natural pigments process.

Alcaligenaceae and Chromatiaceae were previously reported as the specific pollution bioindicators in the receiving river water contaminated by palm oil mill effluent (POME) final discharge. Considering the inevitable sensitivity of bacteria under environmental stresses, it is crucial to assess the survivability of both bacteria in the fluctuated environmental factors, proving their credibility as POME pollution bioindicators in the environment. In this study, the survivability of Alcaligenaceae and Chromatiaceae from facultative pond, algae (aerobic) pond and final discharge were evaluated under varying sets of temperature (25-40°C), pH (pH 7-9) and low/high total suspended solid (TSS) contents of POME collected during low/high crop seasons of oil palm, respectively. Following treatment, the viability status and compositions of the bacterial community were assessed using flow cytometry-based assay and high-throughput Illumina MiSeq, respectively, in correlation with the changes of physicochemical properties. The changes in temperature, pH and TSS indeed changed the physicochemical properties of POME. The functionality of bacterial cells was also shifted where the viable cells and high nucleic acid contents reduced at elevated levels of temperature and pH but increased at high TSS content. Interestingly, the Alcaligenaceae and Chromatiaceae continuously detected in the samples which accounted for more than 0.5% of relative abundance, with a positive correlation with biological oxygen demand (BOD5) concentration. Therefore, either Alcaligenaceae or Chromatiaceae or both could be regarded as the reliable and specific bacterial indicators to indicate the pollution in river water due to POME final discharge despite the fluctuations in temperature, pH and TSS.

The application of zinc oxide nanoparticles (ZnO NPs) in agricultural field is emerging and relatively new. In this work, a simple, cost-efficient, non-toxic and eco-friendly method for the green synthesis of ZnO NPs by Senna occidentalis leaf extract has been described. Techniques used to characterize nanoparticles (NPs) were X-ray diffractometer (XRD), UV visible spectroscopy, Particle size analyzer (PSA), Fourier transform infrared spectroscopy (FTIR) and Scanning electron microscopy (SEM). In this study, green synthesized ZnO NPs at 20-40 mg/l solution was used to prime aged seeds of early flowering homozygous mutant (BM6) of Pusa basmati (Oryza sativa), which enhanced germination performance and seedling vigor significantly as compared to zinc sulphate (ZnSO4) priming and conventional hydropriming. The effect of treatment was analyzed by measuring biophysical and biochemical parameter of germinating rice seeds. The seeds treated with ZnO NPs of 20 mg/l concentration showed more than 50 % stimulation in dry weight, relative water uptake of seeds and radicle length of seedling in comparison to other priming solution and control (hydro-primed). Significant growth was also observed in plumule length and fresh weight of seeds in ZnO NPs at 20 mg/l concentration in comparison to control and other priming treatments. At the same concentration of ZnO NPs, there was 23 % stimulation reported in total soluble sugar content and 45 % stimulation in amylase activity. There was also a substantial increase in antioxidant enzymes i.e. superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) activity. Seed priming represents an innovative user-friendly approach to enhance the germination rate, starch metabolic process and triggered zinc acquisition of rice aged seed as an alternative to the conventional priming method.

Background Jew's ear is an important cultivated edible mushroom. It contains high levels of melanin, carbohydrates, fiber, protein, amino acids, ergosterol, and ascorbic acids, and a very lower level of fat. Various in vitro and in vivo studies have shown that Jew's ear exhibits numerous health-promoting properties. It is known to exhibit antioxidant properties to prevent free radical-induced oxidative damages; balance the blood glucose and lipid levels to alleviate diabetes, hyperlipidemia, and obesity; stimulate macrophage and inflammatory cytokines to prevent inflammation; exhibit anti-cancer properties by stimulating cellular apoptosis and prevent gut microbial dysbiosis by stimulating the growth of healthy gut microbiota. Scope and approach The data regarding A. auricula-judae and its health-promoting effects was obtained through library databases and several electronic searches, such as Google, Google Scholar, PubMed, Scopus, ResearchGate, and journal websites. Key findings and conclusions Our review presented a comprehensive insight into the chemical composition, health-promoting effects, and bioactive mechanisms of A. auricula-judae for the prevention of common diseases.

Growth on solid media as sessile cells is believed to increase the desiccation tolerance of Salmonella enterica. However, the reasons behind increased resistance have not been well explored. In addition, the same effect has not been examined for other foodborne pathogens such as pathogenic Escherichia coli or Listeria monocytogenes. The purpose of this research was twofold: first, to determine the role of oxygenation during growth on the desiccation resistance of S. enterica, E. coli, and L. monocytogenes, and second, to determine the effect of sessile versus planktonic growth on the desiccation resistance of these pathogens. Three different serotypes each of Salmonella, E. coli, and L. monocytogenes were cultured in Trypticase soy broth with 0.6% yeast extract (TSBYE), with (aerobic) shaking or on TSBYE with agar under either aerobic or anaerobic conditions and harvested in the stationary phase. After adding cell suspensions to cellulose filter disks, pathogen survival was determined by enumeration before drying (0 h) and after drying for 24 h. Results showed statistical differences in harvested initial populations before drying. For Salmonella, a correlation was found between high initial population and greater survival on desiccation (P = 0.05). In addition, statistical differences (P ≤ 0.05) between survival based on growth type were identified. However, differences found were not the same for the three pathogens, or between their serotypes. In general, Salmonella and E. coli desiccation resistance followed the pattern of aerobic agar media ≥ liquid media ≥ anaerobic agar media. For L. monocytogenes serotypes, resistance to desiccation was not statistically different based on mode of growth. These results indicate growth on solid media under aerobic conditions is not always necessary for optimal desiccation survival, but may be beneficial when the desiccation resistance of the test serotype is unknown. HIGHLIGHTS

A novel β-glucosidase was purified from pumpkin (Cucurbita moschata) seed by anion exchange chromatography and gel permeation chromatography, and its molecular mass was determined to be 42.8 kDa by gel permeation chromatography. The heterodimeric structure consisting of two subunits, free from disulfide bonds, was determined by native-PAGE analysis followed by zymography. The enzyme was maximally active at pH 4.0 and 70°C, and Vmax, Km, and kcat values were 0.078 units mg−1 protein, 2.22 mM, and 13.29 min−1, respectively, employing p-nitrophenyl-β- d -glucopyranoside as the substrate. The high content of glycine determined by amino acid analysis implies that the enzyme possesses flexible conformations and interacts with cell membranes and walls in nature. Circular dichroism studies revealed that the high stability of the enzyme within the pH range of 2.0–10.0 is due to its reversible pH-responsive characteristics for α-helix–antiparallel β-sheet interconversion.

Food waste is a global issue costing roughly US$ 680 billion annually in industrialized countries. Food spoilage is a major contribution to food waste but artificial food preservatives are often not well accepted by the public. The use of natural alternatives to control spoilage is therefore of interest to the food industry. Lactiplantibacillus plantarum B21, originally isolated from a Vietnamese fermented meat product, produces an antimicrobial bacteriocin peptide which is pH and temperature stable and capable of inhibiting spoilage and pathogenic bacterial strains, including Listeria. In this study the sources of nitrogen and Tween are demonstrated to be essential for growth and bacteriocin production in L. plantarum B21. Conversely, carbohydrate source was not critical for bacteriocin production but was essential for growth. The bacteriocin was found to interact with cell membranes of target microorganisms resulting in cell leakage and subsequent cell death. This study provides insight into the growth conditions needed for the potential development of L. plantarum B21 as a natural biopreservative for food products.

β-1,3-oligoglucoside is widely used as a functional food additive and a physiologically active compound. To efficiently produce β-1,3-oligoglucosides through curdlan hydrolysis, the endo-β-1,3-glucanase gene from Trichoderma harzianum was expressed in Pichia pastoris KM71 for the first time in order to construct a high-yield endo-β-1,3-glucanase strain. Optimal conditions for cell culture and endo-β-1,3-glucanase induction were subsequently determined in shake flask experiments. Recombinant endo-β-1,3-glucanase activity was then evaluated in a 7 L bioreactor, reaching a maximum of 198.57 U/mL after of 118 h fermentation. The recombinant endo-β-1,3-glucanase optimally functioned at pH 5.5 and 50 °C in addition to being stable over a pH range of 4.0–6.0 and a temperature range of 30 °C – 50 °C. In addition, the expressed glucanase could efficiently hydrolyze scleroglucan and schizophyllan to produce branched oligoglucosides exhibiting different degrees of polymerization (DP 3–10) and structures. The high enzyme activity and specificity towards polysaccharides with β-1,3-linked glucose residues as primary chains renders recombinant endo-β-1,3-glucanase an ideal candidate enzyme for the industrial production of oligoglucosides.

Background Black fungi (Auricularia auricula) was known as one of the major mushrooms grown and cultivated all over the world, especially northeast Asia. It has been proved that black fungi had tremendous biological functions, and thus was being remarked to be a potential ingredient for the functional food. Scope and approach This review comprises a comprehensive research for the isolation and characterization of bioactive components in black fungi and their potent physiological activities. Meanwhile, chemical and metabolic methodologies are introduced to clarify the specified mechanism and future challenges. Key findings and conclusions Integrated research results showed that black fungi contain some bioactive components such as polysaccharide, melanin and phenolic. According to origin and isolation method, the structural features of polysaccharide and melanin were significantly different. Health benefits such as antioxidant, hypoglycemic, antitumor and immunostimulation could be exerted with black fungi components and their derivatives. Besides, biological activity and physicochemical property of black fungi enabled the application in the drug delivery and food production. Detailed mechanisms confirmed that oxidative damage removal, immune signal transduction and gut microbiota enhancement supports the promising functionality. However, present research of black fungi still demands the more efficient extraction and purification including subcritical water, ohmic, high hydrostatic pressure (HHP) and two-phase partitioning. Challenges to some health benefits such as gut microbiota-immune, dosage suitable for clinical practice and especially structure-function relationship should also be worthwhile to increase the total functional value of black fungi.

The rhamnogalacturonan acetylesterase gene (named pp1113), encoding a novel cold-adapted rhamnogalacturonan acetylesterase, was cloned from Paenibacillus polymyxa. pp1113 was functionally expressed in Escherichia coli by inducing expression at 20 °C for 16 h. The pH and temperature optimums for the recombinant enzyme were 8.5 and 30 °C, respectively. As an SGNH-type acetylesterase, pp1113 demonstrated high activity against acetyl ester substrates and could maintain approximately 60 % activity at 0 °C.The specific activity of purified pp1113 was 65.95 U/mg (C2). The Km and Vmax values of purified pp1113 were 3.89 mM and 81.07 μmol/min, respectively. We obtained an R232L variant with improved thermal stability and catalytic activity through site-directed mutagenesis. T1/2 of R232L at 50 °C increased from less than 30 min to 6 h when compared to wild-type pp1113, and T1/2 extended to 8 h after immobilization. The R232L mutant improved the conversion rate of 0.1 M cinnamyl alcohol at 40 °C to 94 % in the reaction of synthesis of cinnamyl acetate. After purification, the R232L mutant was immobilized (pH 8.0 phosphate buffer 0.5 M, 20 h at 20 °C, protein loading reached 17 mg/g). This was the first report of the rhamnogalacturonan acetylesterase gene obtained from Paenibacillus polymyxa with detailed enzymatic properties.

The pulp and paper industry in Indonesia is the tenth largest producer in the world, with Acacia and Eucalyptus as the main genera used for production; however, limited publications exist related to Eucalyptus growth models in Indonesia compared with other regions. Time-based models have been developed in which height, stand density, and basal area are predicted based on initial conditions and age. In contrast, a state–space approach utilizes the rate of change of these three state variables. Previous direct comparisons of these two approaches are generally limited. Consequently, the objective of this study was to compare two stand-level growth modeling approaches for Eucalyptus hybrid species on Sumatera (Sumatra) Island using both time-based and state–space methods. Our results indicate that dynamic models using either time-based or state–space approaches are adequate for predicting stand parameters to rotation age. A modified Bazukis matrix indicated that the behavior of both methods produced reliable predictions that were biologically reasonable in terms of stand development; however, the time-based approach provided better performance than the state–space approach on a variety of equivalence tests and goodness-of-fit statistics. Overall, the analysis highlights the advantages and disadvantages of these two commonly used, yet highly contrasting, stand-level growth modeling approaches, which need further consideration and evaluation.

Yeasts are anhydrobiotes that accumulate large amounts of trehalose, which is involved in the vitrification of the cytoplasm during drastic desiccation. The effect of devitrification, which can be induced by the transient exposure of desiccated yeasts to increased humidity or elevated temperature, on the survival of yeast has been studied. A glass transition temperature (Tg)/water activity (aw) diagram of yeast was constructed based on differential scanning calorimetry analysis. The survival rate of yeasts that were equilibrated at different relative humidities (RHs) and temperature values over their Tg range was measured. The results revealed a long period of cell preservation at an intermediate RH (55%), with 100% survival observed after 3 months, a loss of 1.24 log colony-forming units/g recorded after 1 year at 25 °C and full preservation of viability at 75 °C for 60 min and at 100 °C and 12% RH for up to 10 min. These findings led us to conclude that dried yeast can resist low or intermediate RH values and elevated temperatures in the devitrified state. Considering the thermal and humidity fluctuations occurring in the yeast environments, we hypothesized that the supercooled state, which occurs immediately above the Tg after rehydration or heating, is a protective state that is involved in the persistence of yeasts at intermediate humidity levels. KEY POINTS: • Yeast survival for months in a supercooled state is observed at room temperature. • Dried yeasts survive a 10-min exposure to 100 °C in the supercooled state. • The supercooled state is suitable for yeast preservation.

Crop plants have an innate capacity to acclimatize and survive myriad stresses in field conditions. This acclimatization to stress enhances crop stand in field and productivity of plant. Inter alia field crops withstand drought stress (hydropenia) by inducing synthesis or accumulation of osmolytes such as (i) proline and other amino acids, (ii) glycine betaine (GB), (iii) soluble carbohydrates, and (iv) reactive oxygen species (ROS) scavenging system as intrinsic drought antagonizing molecules. Precise in vivo induction of osmolytes and their effect on ROS scavenging system in flax/linseed has not been elucidated. The investigation was carried out to identify a tolerant and susceptible cultivar of flax from a core collection of 53 core accessions and evaluate the role of compatible osmolytes in Linum usitatissimum under hydropenia. We screened eight morphometrically diverse flax genotypes in field under irrigated and un-irrigated condition and classified them as tolerant and susceptible genotypes. Further, we examined the effect of ex-foliar glycine betaine application - a signature molecule involved in drought tolerance, on selected tolerant and susceptible varieties. Our results showed stimulatory impact of glycine betaine on accumulation of ROS scavenging antioxidants, total soluble protein and on its own accumulation. While the ex-foliar application had no inhibitory effect on the growth of plants; accumulation of free proline, amino acids and carbohydrates are inhibited par se in flax. Our findings reveal, flax is a non-accumulator of glycine betaine and exogenous application of glycine betaine enhances its own levels during drought stress.

The use of deep eutectic solvents (DES) with buffer as cosolvent (up to 10 % v/v) leads to low-viscous media in which lipases can perform synthetic reactions, instead of hydrolysis. This paper explores the immobilization of Pseudomonas stutzeri lipase (TL) in cross-linking aggregates (CLEA) to deliver robust derivatives that are active in media like choline chloride – glycerol DES with buffer as cosolvent. While the free TL enzyme was markedly inactive in these media, TL-CLEA derivatives perform esterifications and can be reused several times. Overall, results are consistent with previous experiments reported for other lipases in these DES-water media and confirm that CLEA immobilization turns out a very useful and straightforward alternative for generating active (bio)catalysts for DES-aqueous media systems. Immobilized systems open the possibility of performing continuous processes in low-viscous DES-buffer media.

Aim. To determine the total polyphenols and antioxidant capacity and their correlation with antimicrobial and fungicidal properties in some species of the genus Allium growing in Tajikistan.Materials and methods. The objects of the study were 15 kinds of the Amaryllidaceae family. Antibacterial onions were studied by the disk diffusion method against 4 types of microorganisms (test strains) and 1 type of yeast fungus. In the presence of total polyphenols, the content in μg of Gallic acid (GA) per ml of plant extract was measured. To determine the antioxidant activity, a solution of ABTS K2S2O8 was used. Trolox was used as a standard.Results. The maximum content of total polyphenols is characterized by ethanol extracts of A. Carolinianum, A. altaicum, A. elatum, A. oschaninii, A. ramosum, A. hymenorhisum. At the same time, their lowest amount was found in the bulbs of A. longicuspis. A. shoenoprasum, which is statistically significantly lower than in all the above-listed species of this plant group. The extracts from the stems of A. carolinianum, A. elatum, and A. pamiricum had the highest antioxidant effect. There is a direct weak correlation between biologically active components with antimicrobial and fungicidal activity in A. oshaninii and A. suworowii, depending on the altitude gradient.Conclusions. The results allow suggesting that the above-ground and underground parts of some wild-growing endemic species of the genus Allium have different degrees of antioxidant activity due to total polyphenols. The content of total polyphenols and the antioxidant capacity of extracts obtained from various plants in different in large ranges. In some cases, there are direct correlations between the content of polyphenols, antioxidant properties, antimicrobial and fungal properties.

Introduction: One of plants that may have therapeutic potential is the Cymbopogon schoenanthus (L.) Spreng, also known as camel grass. Aim: This review aims to investigate and gather comprehensive information about camel grass plants and their potential to be developed into a nanotechnology drug delivery system. Methods: This review examined a variety of online literature. Results: It was found that camel grass contains essential oil such as piperitone. Piperitone is efficacious as an antioxidant, antimicrobial and anti-inflammatory, in addition to other properties . The development of camel grass essential oil into lipid-based nanotechnology preparations can improve its bioavailability, solubility, and stability, thereby improving its potential effectiveness.

The growth of man-caused load on natural objects, including surface waters, gave impetus to the studying for issue of water pollution in Ukraine by various pollutants. Especially relevant for us are the studies of fishery reservoirs as a direct environment for growing quality aquaculture products. Among the toxic compounds contained in reservoirs, one of the first places is occupied by heavy metals. On the one hand, heavy metals, as pollutants in natural waters, pose a great danger because even in relatively small concentrations they can adversely affect aquatic organisms. On the other hand, in microquantities most heavy metals (except mercury, cadmium and lead) are a natural and even essential component of living cells of aquatic organisms, including fish [17,19].Zn, Mn, Fe, Cu, Pb, Co, Ni, Cd in the organs and tissues of the bighead carphybrid form, which was cultivated in ponds from highly urbanized territory, are distributed heterogeneously and their level of content depends not only on the physical and chemical characteristics of the aquatic environment or this heavy metals biochemical activities force in fish organisms, but also on functional features of concrete organs and tissues of the latter. Results of our investigations showed that above mentioned heavy metals were characterised by high levels of content in the organs and tissues of hybrid silver x bigheadcarp. Highest content in organism of hybrid are lead, cobalt, and cadmium. Highest concentration of heavy metals in our investigations was recorded in gills, because it is the first defence line of fish organism from environmental pollutants.

International audience; Beardless barb is a common fish species used in fermentation of fish paste Ka-pi-plaa. Autolytic profile of beardless barb muscle showed the maximum autolysis was at 50 °C, at both acidic and alkaline pH values. With augmentation concentration of NaCl, autolytic activity slightly decreased. Endogenous proteinases isolated from fish muscle in crude extract forms were also characterised. The acidic proteinases had optimum activity at pH 3.0 and 50°C, and they showed higher proteolytic activity than the alkaline proteinases which were optimally active at pH 9.0 and 50 °C. Proteinases in peak at pH 3.0 were inhibited by pepstatin A, but those in peak at pH 9.0 were highly inhibited by PMSF, TLCK and soybean trypsin inhibitor, suggesting that both aspartic and serine proteinases were existed in beardless barb muscle. The proteinases were stable in pH range of 2.0-5.0 but unstable at the temperatures higher than 40 °C. NaCl suppressed the proteolytic activity, ATP activated the proteinase activity, while CaCl2, MgCl2 and CoCl2 exhibited no influence on the activity. The results implied that cathepsin D is the predominant proteinase responsible for autolysis in beardless barb. The findings were useful to improve the processing and qualities of Ka-pi-plaa product using beardless barb as raw material.