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Enzymes are biological catalysts that play a key role in the food industry, responsible for desirable and undesirable chemical reactions. These compounds can occur spontaneously in food products or even be incorporated on purpose. The main enzyme classes (oxidoreductase, transferase, hydrolase, lyase, isomerase, and ligase) and their subclasses can be obtained from several sources, presenting numerous applications in foods such as inhibition of microorganisms, insertion of aromas, improvement of physico-chemical properties, decreased candy crystallization, meat tenderization, antioxidants, indicators of heat treatment, and so on. Each enzyme acts effectively under specific conditions of pH, temperature, concentration, and water activity. Enzymatic immobilization techniques have been studied to minimize adverse conditions that enzymes are subjected to during food processing. Among the immobilization technique, the enzymes can be immobilized to the packages, being known as enzymatic active packages, the packages allow the enzymes to be migrated to the product or trapped, increasing the shelf life of the products, this technique being innovative both for the packaging market and for the enzyme market. Thus, the main enzymes and their applications in food were briefly discussed in this review article, as well as the main techniques of immobilization and insertion of these compounds in active packaging.

A Plackett-Burman factorial design with 15 experiments was conducted to evaluate the influence of seven factors on lipases production by Fusarium solani. The factors investigated were peptone, tryptone, yeast extract, calcium chloride, potassium phosphate, magnesium sulfate, and copper sulphate. Five fixed variables (cotton oil, pH, temperature, agitation, and time) were maintained and as a response to the enzymatic activity. The concentration of tryptone, calcium chloride, and magnesium sulphate had a significant effect (p < 0.10) on lipase production and was studied consecutively through a complete DCCR (central rotational compound design), to optimize lipase production of the fungi F. solani. After optimization using DCCR, maximum lipolytic activities of 24.84 U/ml were obtained with the use of 10 g.L-1 tryptone, 3.50 g.L-1 calcium chloride and 0.50 g.L-1 magnesium sulfate, 1 g.L-1 potassium phosphate and 1% soybean oil. The statistical model showed a correlation of 85.67% with the experimental data. The biochemical characterization of lipase showed that the enzyme has a better performance at pH 7 at a temperature of 40 °C, where the statistical model had a correlation of 94.15% with the experimental data. In this way, lipases produced by F. solani have potential for application and use in biodiesel production. Se realizó un diseño factorial de Plackett-Burman con 15 experimentos para evaluar la influencia de siete factores en la producción de lipasas por Fusarium solani. Los factores investigados fueron peptona, triptona, extracto de levadura, cloruro de calcio, fosfato de potasio, sulfato de magnesio y sulfato de cobre. Se mantuvieron cinco variables fijas (aceite de algodón, pH, temperatura, agitación y tiempo) y como respuesta a la actividad enzimática. La concentración de triptona, cloruro de calcio y sulfato de magnesio tuvo un efecto significativo (p < 0.10) sobre la producción de lipasa y fue estudiada consecutivamente a través de un DCCR completo (diseño de compuestos rotacionales centrales), para optimizar la producción de lipasa del hongo F. solani. Después de la optimización con DCCR, se obtuvieron actividades lipolíticas máximas de 24,84 U/ml con el uso de 10 g.L-1 de triptona, 3,50 g.L-1 de cloruro de calcio y 0,50 g.L-1 de sulfato de magnesio, 1 g.L-1 de fosfato de potasio y 1% de aceite de soja. El modelo estadístico mostró una correlación del 85,67% con los datos experimentales. La caracterización bioquímica de la lipasa mostró que la enzima tiene un mejor desempeño a pH 7 a una temperatura de 40 °C, donde el modelo estadístico tuvo una correlación de 94.15% con los datos experimentales. De esta forma, las lipasas producidas por F. solani tienen potencial de aplicación y uso en la producción de biodiesel. Um planejamento fatorial Plackett-Burman com 15 experimentos foi conduzido para avaliar a influência de sete fatores na produção de lipases por Fusarium solani. Os fatores investigados foram peptona, triptona, extrato de levedura, cloreto de cálcio, fosfato de potássio, sulfato de magnésio e sulfato de cobre. Cinco variáveis ​​fixas (óleo de algodão, pH, temperatura, agitação e tempo) foram mantidas e como resposta à atividade enzimática. A concentração de triptona, cloreto de cálcio e sulfato de magnésio teve efeito significativo (p < 0,10) na produção de lipase e foi estudada consecutivamente através de um DCCR completo (delineamento de composto rotacional central), para otimizar a produção de lipase do fungo F. solani. Após otimização com DCCR, atividades lipolíticas máximas de 24,84 U/ml foram obtidas com o uso de 10 g.L-1 de triptona, 3,50 g.L-1 de cloreto de cálcio e 0,50 g.L-1 de sulfato de magnésio, 1 g.L-1 de fosfato de potássio e 1% de óleo de soja . O modelo estatístico apresentou uma correlação de 85,67% com os dados experimentais. A caracterização bioquímica da lipase mostrou que a enzima tem um melhor desempenho em pH 7 na temperatura de 40 °C, onde o modelo estatístico teve uma correlação de 94,15% com os dados experimentais. Desta forma, as lipases produzidas por F. solani apresentam potencial para aplicação e uso na produção de biodiesel.

Background Omega fatty acids are a family of polyunsaturated fats associated with several health benefits. Lipases are enzymes with potential application in several food processes such as flavor and aroma, surfactants and formulations for the dairy and bakery industries. In this study, single cell oil and lipase production by Candida viswanathii CCR8137 were evaluated simultaneously from renewable carbon sources under nitrogen limitation. Methods Enzyme and single cell oil were obtained in submerged cultivations supplemented with triolein, tributyrin, corn oil, sunflower oil, canola oil and olive oil. The effects of glucose on lipid accumulation, fatty acid profile, enzyme production and cell morphology were also evaluated. Results The highest lipid accumulation (44.5%, w/w) was obtained from triolein, whereas olive oil was the best inducer of lipase synthesis (26.8 U/mL). Nitrogen limiting cultivations were a key parameter for an organic source which showed higher lipid accumulation and enzyme production than the tested inorganic nitrogen source. Glucose was a poor inducer of lipase synthesis, though increased values of lipid accumulation were observed from this carbon source with a maximum of 63.1% (w/w). The fatty acid profile of lipids produced by C. viswanathii CCR8137 showed a high content of omega-9 fatty acid (C18:1 n-9). The addition of glucose to the culture media resulted in the synthesis of essential fatty acids: vaccenic, linolenic and eicosadienoic acids. Conclusions Therefore, C. viswanathii CCR8137 strain can be considered as an oleaginous yeast able to accumulate high concentrations of intracellular lipids, which are potential additives for food industry applications as well as being able to simultaneously synthesize high yields of lipase.

Research background. Microbial β-fructofuranosidases are widely employed in food industry to produce inverted sugar or fructooligosaccharides. In this study, a newly isolated Aspergillus carbonarius PC-4 strain was used to optimize the β-fructofuranosidase production in a cost-effective process and the sucrose hydrolysis was evaluated to produce inverted sugars. Experimental approach. Optimization of nutritional components of culture medium was carried out using simplex lattice mixture design for 72 and 120 h at 28 °C. One-factor-at-a-time methodology was used to optimize the physicochemical parameters. Crude enzyme was used for sucrose hydrolysis at different concentrations. Results and conclusions. The optimized conditions of enzyme production were achieved from cultivations containing pineapple crown waste (1.3%, m/V) and yeast extract (0.3%, m/V) after 72 h with an enzyme activity of 9.4 U/mL, obtaining R2=91.85%, R2adjusted=85.06%, highest F-value (13.52) and low p-value (0.003). One-factor-at-a-time used for optimizing the physicochemical conditions showed optimum temperature (20 °C), pH (5.5), agitation (180 rpm) and time course (72 h) with a 3-fold increase of enzyme production. The invertase-induced sucrose hydrolysis showed the maximum yield (3.45 mmol of reducing sugars) using 10% of initial sucrose concentration. Higher sucrose concentrations caused the inhibition of invertase activity, possibly due to the saturation of substrate or formation of sucrose aggregates, making it difficult for the enzyme to access sucrose molecules within the created clusters. Therefore, a cost-effective method was developed for the invertase production using agroindustrial waste and the produced enzyme can be used efficiently for inverted sugar production at high sucrose concentration. Novelty and scientific contribution. This study presents an efficient utilization of pineapple crown waste to produce invertase by a newly isolated Aspergillus carbonarius PC-4 strain. This enzyme exhibited a good potential for inverted sugar production at high initial sucrose concentration, which is interesting for industrial applications., Pozadina istraživanja. Mikrobne β-fruktofuranozidaze imaju široku primjenu u prehrambenoj industriji, i to u proizvodnji invertnog šećera ili fruktooligosaharida. U ovom je radu korišten novi soj gljivice Aspergillus carbonarius PC-4 za optimiranje ekonomski isplativog postupka proizvodnje β-fruktofuranozidaze, te je ispitana hidroliza saharoze radi dobivanja invertnih šećera. Eksperimentalni pristup. Sastav hranjive podloge optimiran je primjenom simpleks-metode tijekom 72 i 420 h pri 28 °C. Jednofaktorskom su metodom optimirani fizikalno-kemijski uvjeti proizvodnje enzima. Sirovi je enzim korišten za hidrolizu saharoze pri različitim koncentracijama. Rezultati i zaključci. Optimirani su uvjeti proizvodnje enzima dobivenog uzgojem na podlozi s krunom ploda ananasa (1,3 % m/V) i ekstraktom kvasca (0,3 % m/V) nakon 72 h, pri čemu je aktivnost enzima iznosila 9,4 U/mL, s R²=91,85 %, R²adj=85,06 %, najvišom F-vrijednosti (13,52) i niskom p-vrijednosti (0,003). Jednofaktorskom su metodom određeni sljedeći optimalni fizikalno-kemijski uvjeti: temperatura od 20 °C; pH-vrijednost od 5,5; brzina miješanja od 180 okretaja u minuti i trajanje postupka od 72 h, pri čemu je proizvedena trostruko veća količina enzima. Najveći prinos reducirajućih šećera pri hidrolizi saharoze pomoću invertaze s početnom količinom od 10 % saharoze bio je 3,45 mmol. Veće količine saharoze inhibirale su aktivnost invertaze, vjerojatno zbog zasićenja podloge ili formiranja nakupina šećera koje su enzimu otežale pristup molekulama saharoze. Zaključeno je da je razvijen ekonomski isplativ postupak proizvodnje invertaze iz agroindustrijskog otpada, a da se dobiveni enzim može učinkovito upotrijebiti za dobivanje invertnog šećera pri većim količinama saharoze. Novina i znanstveni doprinos. U radu je prikazan djelotvoran način iskorištenja krune ploda ananasa kao biootpada u proizvodnji invertaze s pomoću novog soja gljivice Aspergillus carbonarius PC-4. Dobiveni je enzim pokazao potencijal za proizvodnju invertnog šećera pri velikoj početnoj količini saharoze, što je korisno za industrijsku primjenu.

Os alimentos transgênicos vêm ganhando mais espaço na agricultura brasileira, principalmente na região do Cerrado. O objetivo deste trabalho foi analisar a percepção e aceitabilidade da população tocantinense a respeito dos alimentos transgênicos. Aplicou-se questionários contendo 29 questões para 119 pessoas residentes em Gurupi-TO e outras cidades da região sul do Estado. Os questionários foram aplicados nos anos de 2016 a 2018 e analisados pelo método estatístico descritivo. Obteve-se um grau de confiança de 92,9% e margem de erro de 3,15 pontos percentuais. Observou-se que, 74% dos entrevistados conheciam a definição de alimentos transgênicos, 69% afirmaram que optariam por um alimento não transgênico. No entanto, 74% afirmam não leem o rótulo para verificar se o alimento é transgênico. Quanto ao consumo de alimentos transgênicos, 89% acreditam que a população consome sem conhecimento prévio. Portanto, nota-se a necessidade de inserção da população nos debates referentes aos transgênicos, bem como uma maior divulgação científica e midiática a respeito do tema para sanar as dúvidas da população sobre assunto.

Extracellular fructosyltransferase (FTase, E.C.2.4.1.9) from Aspergillus oryzae IPT-301 was immobilized on silica gel by adsorption and biochemically characterized aiming at its application in the transfructosylation reaction of sucrose for the production of fructooligossaccarides (FOS). The transfructosylation activity (AT) was maximized by the experimental design in function of the reaction pHs and temperatures. The AT of the immobilized enzyme showed the kinetics behavior described by the Hill model. The immobilized FTase showed reuse capacity for six consecutive reaction cycles and higher pH and thermal stability than the soluble enzyme. These results suggest a high potential of application of silica gel as support for FTase immobilization aiming at FOS production.

A goma xantana é um biopolímero microbiano de grande importância na indústria alimentícia. A produção desse polissacarídeo é influenciada diretamente pelas condições de cultivo, linhagens e substratos empregados como sacarose e glicose, o que tornam o processo oneroso. A busca por linhagens que apresentem boa relação quantidade/qualidade e por substratos que diminuam o custo de produção, como por exemplo os resíduos agroindustriais, tem sido foco de pesquisas recentes. O presente estudo visou produzir goma xantana utilizando o soro de leite em substituição parcial da sacarose no meio de fermentação, na proporção de 1:1. Utilizou-se a cepa selvagem de Xanthomonas sp. MN37.2, isolada de tecidos foliares de mangueira. A Fermentação controle foi conduzida com meio contendo 50g/L de sacarose. As fermentações foram realizadas à 28°C, à 180 rpm por 96 horas. O pH decaiu de 6,54 para 5,18 durante o processo fermentativo da cepa e a população de células viáveis aumentou de 10,15 log UFC/mL para 11,69 log UFC/mL. O rendimento de produção da goma na fermentação com adição de soro de leite foi de 62,13% quando comparado à fermentação com sacarose como única fonte de carbono, sendo 30,35 g/L e 48,85 g/L, respectivamente. Portanto, o soro de leite demonstra potencial para ser empregado como substrato de baixo custo na obtenção de goma xantana por bactérias do gênero Xanthomonas. Entretanto, estudos de otimização da formulação do meio de fermentação, bem como controle das etapas de recuperação da goma devem ser realizados afim de viabilizar a utilização em escala industrial desse subproduto.

The aim of this work was to evaluate the use of different agro-industrial residues for the production of xylanase by Aspergillus sp. ART 100.1, as well as analyzing the biochemical properties of the enzyme. Agroindustrial residues malt bagasse, pineapple crown, açaí bagasse and soybean husk present in large quantities in the Tocantins region were used to evaluate the production of xylanase. Cultivation conditions for xylanase production were evaluated in submerged and solid-state cultivation. The highest production of xylanase in submerged cultivation was obtained using soybean husk residue (23.60 U/mL), while, for solid-state cultivation conditions, the highest production of xylanase was obtained with malt bagasse (110.00 U/g). The effect of additives to the culture medium was also evaluated, with the best result for the use of the xylose additive in the pineapple crown in solid-state cultivation. The enzyme produced in solid-state cultivation was characterized in terms of pH and temperature. The optimum activity pH was observed at 5.0 and for temperature at 55 °C. The xylanase was stable in a pH range between 4 and 5 and retained 50% of its activity at 45 °C after 110 minutes. The Aspergillus sp. ART 500.1 presents potential for the production of xylanase using agro-industrial residues, enabling the development of bioprocesses for the scaling of production.

The development of microbial enzymes is of great importance in biochemical process industries. Among the enzymes of industrial importance, cellulolytic and xylanolytic enzymes stand out, which are responsible for the degradation of lignocellulosic material that is usually discarded by the agro-industrial sector. In order to produce the enzymes CMCase, β-glycosidase, xylanase and β-xylosidase, the fungus Fusarium oxysporum was used in liquid fermentation using pre-treated pineapple crown biomass as a carbon source. In order to optimize the production of these enzymes, cultures were carried out for 168 hours, 30 °C at 180 rpm. The biomass underwent a pre-treatment in two stages, the first being submitted to 121 °C for 7 min, while the second was added diluted sulfuric acid to 1%, in the proportion of 1:5 (m/v), at 121 °C for 27 minutes. The partial characterization of the crude extract showed a temperature of 50 ºC as the most satisfactory for the action of these enzymes, and the ideal pH range ranged from 3.5 to 5.5. The enzymatic hydrolysis of pineapple crown, with and without pretreatment, was performed using lyophilized crude extract (CMCase = 9.78 U/mL, β-glucosidase = 10.86 U/mL, xylanase = 27.90 U/mL and β-xylosidase = 0.018 U/ml). An experimental design of the Central Composite Rotational Design was carried out. It was possible to evaluate the effect and influence of pH, biomass concentration and incubation time on the release of glucose in the medium (R2>0.85); pH and biomass (%) had a significant effect on glucose release.

This study aimed to optimize the production of xylanase, under submerged conditions using Fusarium oxysporum. Firstly, it was carried out the selection of the best source of carbon from agricultural waste, such as soybean husk, cassava peel, pineapple crown, corn straw, rice husk, bacaba, barley bagasse and, corncob. They were milled and sieved with a maximum granulation of 1 mm. The cultures were carried out by 132 hours 30 °C and 180 rpm. Among the agroindustrial residues, corn straw was the one that stood out with a greater enzymatic activity of 12.6 U/mL. In the second step, it was used Plackett-Burman design to screen the nutrients sources important to xylanase production. Thus, independent variables significant were urea and MgSO4. These variables selected by Plackett-Burman were then used in a Central Composite Rotational Design present activity of 26.6 U/mL and the predicted was 34.5 U/mL. So, the xylanase production by F. oxysporum can be optimize using corn straw, a low-cost waste found in large quantities.

Lipases are an important group of enzymes that catalyze the hydrolysis of triacylglycerol and there are many industrial applications. The aim of this work was to produce the lipase by the yeast Candida viswanathii using solid state culture with agro-industrial wastes (barley bagasse, corn husk, corncob, soybean seed coat and soybean husk). The biomass pretreatment methods were evaluated, as well as the media supplementation with nitrogen and mixing substrates. Also, the efficiency of olive oil and poultry fat was evaluated on the induction of lipase production, followed by the scale-up from 20 g to 100 g. The enzyme activities in the cultures without pretreatement were higher when soybean seed coat supplemented with both olive oil (7.06 U/gss) and poultry fat (8.40 U/gss) were used. However, the pretreated substrates did not demonstrate a satisfying induction of lipolytic activity. From the nitrogen sources, yeast extract showed an increase of approximately twice the original production with both olive oil (18.12 U/gss) and poultry fat (15.98 U/gss) supplementation. On the scale-up step, the results demonstrated that, for the 20 g culture, the best lipase production was observed on the 7th day (33.52 U/gss), while for the 100 g culture the highest lipase activity was after 5 days (17.88 U/gss). The cultivation of ground soybean skin without pretreatment supplemented with yeast extract as a source of nitrogen, with fresh barley bagasse and poultry fat was the best combination.

Made available in DSpace on 2022-04-28T19:29:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-05-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Agroindustrial wastes is an interesting opportunity for enzymes production, reducing operational costs and strengthening the biorefineries concept. The conversion of lignocellulosic waste to fermentable sugars is current challenge for biorefineries industries. Enzyme cocktail used in delignification and saccharification platform are produced by a combinatory of several lignocellulolytic enzymes. The aim of this study was to develop filamentous fungi compatible-consortia isolated from pineapple wastes for cellulose-degrading enzymes production using pineapple crown waste in solid-state cultivation and its potential for saccharification of pineapple crown waste. Isolates were screened in agar-plate and in solid-state cultivation for cellulases, xylanase and pectinases productions. Six compatible consortia of Trichoderma strains with Aspergillus niger or Pleurotus ostreatus were evaluated for SSC using pineapple crown. All consortia increased the enzymes production compared to monoculture. Consortia of Trichoderma asperellum PEC-6 and P. ostreatus increased 1.60-fold the xylanase and 1.42-fold the β-glucosidase productions. T. asperellum PEC-17 and A. niger increased 1.16-fold endoglucanase and 1.28-fold pectinase. Saccharification of pineapple crown waste using enzyme cocktail produced by filamentous fungi consortia increased the total reducing sugar released in 12.50–13.93% compared to enzymes produced by monocultures. This study provides an alternative model to the cocktail enzyme production by mixed cultures development with lower cost of on-site enzyme manufacture by use of agroindustrial wastes eliminating an enzyme blended step. Moreover, the procedure used in this work can be potentially cost saving and environmentally friendly and should be explored on other bioenergy feedstocks and feed production. Graphic Abstract: [Figure not available: see fulltext.] Bioprocess Engineering and Biotechnology Federal University of Tocantins Post-Graduate Food Science and Technology Federal University of Tocantins Habite - Biotechnology-Based Incubator Companies Federal University of Tocantins Biochemical and Microbiology Department Univ. Est. Paulista Federal University of Bahia Vitória da Conquista Embrapa Agroenergia CAPES: Finance Code 001

The selection of microorganisms with the greatest potential of composer enables to increase efficiency and reduce the time of the composting process. The isolation of fungi has received special attention because they are the main responsible for the degradation of substances of difficult decomposition during the composting process. This review presents a survey of fungi with cellulolytic enzymatic potential, the methods of isolation and screening of these fungi and the methods for determining the potential enzymatic assay. The fungi that decompose rich cellulose residues are the main decomposing agents of ruminal residue, have a diversity of enzymes with degradation capacity of complex organic compounds, such as cellulose, hemicellulose, acids aromatics, and some proteins. Thus, with the indication of fungi that increase the composting efficiency, specific glasses can be developed for the transformation of this type of residue.

In this study, Penicillium sp. section Gracilenta CBMAI 1583 was used to produce lipase under submerged conditions. The enzyme was purified and the biochemical properties of both the crude and purified enzymes were evaluated. Maximum lipase production (1.62 U mL−1) was obtained using olive oil 0.5% (w v−1) after 72 h of cultivation, representing a 90% increase in the lipase initially produced. The enzyme was purified using hydrophobic interaction chromatography (phenyl Sepharose) under conditions which allowed its interfacial activation. The partially purified sample showed an enzyme with esterase activity (65.4 kDa) on α- and β-naphthyl acetate and other with lipase activity (52.9 kDa) on octyl oleate. Optimum activity of crude and purified lipase was observed at pH 4.0 and 70 °C. The purified lipase was activated by NaCl, BaCl2, NH4Cl, MnSO4 and MgSO4; it also presented high stability in organic solvents such as hexane, 2.2.4-trimethylpentane, acetone, DMSO and toluene. Maximum enzyme activity was observed with p-nitrophenyl decanoate as substrate; and the enzyme kinetics showed to be directly affected by Triton X-100. The enzyme shows potential application in processes that operate in acid pH, such as treatment of dairy and industry effluents, resolution of esters in the pharmaceutical industry or in the food industry, as well in synthesis reaction under non-aqueous conditions., The authors gratefully acknowledge to São Paulo Research Foundation - FAPESP, Brazil, for the scholarship granted to the first author and Spanish Ministry of Science and Innovation - MICINN, Spain (Project BIO-2012–36861).

Polyhydroxyalkanoates (PHAs) are intracellular microbial polyesters synthesized by many species of Bacteria and Archaea, generally under nutrient limitation and excess of carbon source as storage granules of energy and also conferring stress resistance to prokaryotes. PHAs are naturally occurring, and the choice of microbial strains and carbon sources for cell cultivations results in biopolymers with different physicochemical properties. Cupriavidus necator is a bacterial strain able to synthesize a high crystalline polymer, poly(3-hydroxybutyrate), the most common of all PHAs; whereas Pseudomonas species produce PHAs with medium chain-length monomers, which exhibit elastomeric properties as natural rubbers. This variety of monomer composition associated with high biocompatibility and biodegradability makes PHAs also feasible for compostable applications and meet the standard specification for marine degradability. Due to their high versatility as natural chemicals and their ecologically friendly properties, PHAs have drawn attention for a wide range of applications from which the PHA nanocomposites have become a promising alternative to petrochemical polymers for nanoscale applications, especially those involved in biomedical materials. This review chapter introduces PHAs from their microbial synthesis, polymer structure to recent applications, with special focus on nanoscience applications. PHA as scaffolds for tissue engineering, nanofibers and particles for catalyst immobilization and drug delivery systems are topics addressed in this chapter. Finally, the future prospects and economical challenges to be overcome by the PHA industry are discussed in this review chapter from a green chemistry background.

In this work, it was determined the influence of temperature, pH and storage time on the enzymatic activity and stability of an extracellular fructosyltransferase (FTase E.C.2.4.1.9) from Aspergillus oryzae IPT-301 produced by submerged fermentation. The thermodynamic parameters showed a tendency for increasing enzyme denaturation with the rise in temperature. The maximum transfructosylation activity was obtained at the incubation pH 5.5. During storage at 4 °C, the transfructosylation activity decreased, whereas the hydrolytic activity increased, especially in the first nine hours, a time in which the enzyme presented 45.6% of its initial transfructosylation activity. These results contributed to the improvement of the conditions of storage, immobilization and use of the soluble fructosyltransferases (FTase) in fructooligosaccharide (FOS) production. Resumo Neste trabalho foi determinada a influência da temperatura, do pH e do tempo de armazenamento na estabilidade enzimática de frutosiltransferase (E.C.2.4.1.9) extracelular de Aspergillus oryzae IPT-301, produzida por fermentação submersa. Os parâmetros termodinâmicos mostraram tendência de aumento da desnaturação da enzima com o aumento da temperatura. A atividade de transfrutosilação máxima foi obtida para a enzima incubada em pH 5,5. Durante o armazenamento a 4 °C, a atividade de transfrutosilação diminuiu, enquanto a atividade hidrolítica aumentou, principalmente nas primeiras nove horas, período em que a enzima apresentou 45,6% de sua atividade inicial de transfructosilação. Esses resultados contribuem para a melhoria das condições de armazenamento, imobilização e utilização das frutosiltransferases solúveis na produção de frutooligossacarídeos.

Isolating high quality RNA is a limiting factor in molecular analysis, since it is the base for transcriptional studies. The RNA extraction method can directly affect the RNA quality and quantity, as well as, its overall cost. The industrial importance of the yeast genus Candida in several sectors comes from their capacity to produce Lipases. These enzymes are one of the main metabolites produced by some Candida species, and it has been shown that Candida yeast can biodegrade petroleum hydrocarbons and diesel oil from biosurfactants that they can produce, a feature that turns these organisms into potential combatants for bioremediation techniques. Thus, this study aimed to determine an efficient method for isolating high quality RNA from Candida viswanathii biomass. To achieve this aim, three different RNA extraction methods, TRIzol, Hot Acid Phenol, and CTAB (Cetyltrimethylammonium Bromide), were tested. The three tested methods allowed the isolation of high-quality RNA from C. viswanathii biomass and yielded suitable RNA quantity for carrying out RT-qPCR studies. In addition, all methods displayed high sensitivity for the expression analysis of the CvGPH1 gene through RT-qPCR, with TRIzol and CTAB showing the best results and the CTAB method displaying the best cost-benefit ratio (US$0.35/sample).

Polyhydroxyalkanoate (PHA) bioplastic was synthesized by Burkholderia glumae MA13 from carbon sources and industrial byproducts related to sugarcane biorefineries: sucrose, xylose, molasses, vinasse, bagasse hydrolysate, yeast extract, yeast autolysate, and inactivated dry yeast besides different inorganic nitrogen sources. Sugarcane molasses free of pre-treatment was the best carbon source, even compared to pure sucrose, with intracellular polymer accumulation values of 41.1–46.6% cell dry weight. Whereas, xylose and bagasse hydrolysate were poor inducers of microbial growth and polymer synthesis, the addition of 25% (v/v) sugarcane vinasse to the culture media containing molasses was not deleterious and resulted in a statistically similar maximum polymer content of 44.8% and a maximum PHA yield of 0.18 g/g, at 34°C and initial pH of 6.5, which is economic and ecologically interesting to save water required for the industrial processes and especially to offer a fermentative recycling for this final byproduct from bioethanol industry, as an alternative to its inappropriate disposal in water bodies and soil contamination. Ammonium sulfate was better even than tested organic nitrogen sources to trigger the PHA synthesis with polymer content ranging from 29.7 to 44.8%. GC-MS analysis showed a biopolymer constituted mainly of poly(3-hydroxybutyrate) although low fractions of 3-hydroxyvalerate monomer were achieved, which were not higher than 1.5 mol% free of copolymer precursors. B. glumae MA13 has been demonstrated to be adapted to synthesize bioplastics from different sugarcane feedstocks and corroborates to support a biorefinery concept with value-added green chemicals for the sugarcane productive chain with additional ecologic benefits into a sustainable model.

The demand for functional foods is increasing each year because consumers are gaining awareness about the importance of a healthy diet in the proper functioning of the body. Probiotics are among the most commonly known, commercialized, and studied foods. However, the loss of viability of probiotic products is observed during their formulation, processing, and storage. This study aimed to investigate the co-encapsulation of two Lactobacillus paracasei probiotic strains (LBC81 and ELBAL) with fructooligosaccharides (FOS) in a calcium alginate matrix using extrusion technology with gelatin as a coating material. The viability of the strains under gastrointestinal conditions and in storage at low temperature was also assessed. An immobilization yield of more than 59% was observed for both bacterial strains. Exposure to 2% biliary salts led to a decrease in the viability of free cells in the two L. paracasei strains, whereas the viability of microencapsulated cells increased up to 47%. After 35 days of storage at 4°C, the population of free cells was reduced, but microencapsulated cells remained stable after storage at low temperature. LBC81 bacteria microencapsulated with 1.5% FOS coated with gelatin were the most resistant to the stressful environments tested. Therefore, these results showed that co-encapsulation with FOS in a calcium alginate matrix coated with gelatin improved L. paracasei survival and may be useful for the development of more resistant probiotics and new functional foods. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s42770-021-00484-5.

SUMMARY Research background Microbial β-fructofuranosidases are widely employed in food industry to produce inverted sugar or fructooligosaccharides. In this study, a newly isolated Aspergillus carbonarius PC-4 strain was used to optimize the β-fructofuranosidase production in a cost-effective process and the sucrose hydrolysis was evaluated to produce inverted sugars. Experimental approach Optimization of nutritional components of culture medium was carried out using simplex lattice mixture design for 72 and 120 h at 28 °C. One-factor-at-a-time methodology was used to optimize the physicochemical parameters. Crude enzyme was used for sucrose hydrolysis at different concentrations. Results and conclusions The optimized conditions of enzyme production were achieved from cultivations containing pineapple crown waste (1.3%, m/V) and yeast extract (0.3%, m/V) after 72 h with an enzyme activity of 9.4 U/mL, obtaining R2=91.85%, R2adjusted=85.06%, highest F-value (13.52) and low p-value (0.003). One-factor-at-a-time used for optimizing the physicochemical conditions showed optimum temperature (20 °C), pH (5.5), agitation (180 rpm) and time course (72 h) with a 3-fold increase of enzyme production. The invertase-induced sucrose hydrolysis showed the maximum yield (3.45 mmol of reducing sugars) using 10% of initial sucrose concentration. Higher sucrose concentrations caused the inhibition of invertase activity, possibly due to the saturation of substrate or formation of sucrose aggregates, making it difficult for the enzyme to access sucrose molecules within the created clusters. Therefore, a cost-effective method was developed for the invertase production using agroindustrial waste and the produced enzyme can be used efficiently for inverted sugar production at high sucrose concentration. Novelty and scientific contribution This study presents an efficient utilization of pineapple crown waste to produce invertase by a newly isolated Aspergillus carbonarius PC-4 strain. This enzyme exhibited a good potential for inverted sugar production at high initial sucrose concentration, which is interesting for industrial applications.

O mercado global de alimentos prevê taxas contínuas de crescimento entre os anos de 2019 a 2024. Este crescimento se deve à demanda por personalização e inovação na variedade, conveniência e qualidade dos produtos, que pressionam a indústria de alimentos e bebidas a diferenciar seus produtos por meio de processos inovadores. Essa tendência traduz também, a necessidade de garantir a segurança alimentar e a qualidade nutricional dos produtos alimentícios. O desenvolvimento da inovação, tecnologia e sustentabilidade no setor de alimentos requer o emprego de áreas multidisciplinares não somente para desenvolver o setor industrial, mas também para o desenvolvimento da agricultura familiar e dos pequenos e médios produtores que tem como objetivo a produção de alimentos e bebidas regionais e artesanais. Com o objetivo de discutir as tendências no setor de alimentos, o tema “Inovação tecnológica e sustentabilidade: bases para o futuro da indústria de alimentos” foi abordado no evento XV Semana Acadêmica de Engenharia de Alimentos (SEMEALI) e II Congresso Tocantinense de Ciência e Tecnologia de Alimentos (CTOCTA), realizado entre os dias 04 e 08 de novembro de 2019, na Universidade Federal do Tocantins, no munícipio de Palmas. O evento organizado pelo curso de Engenharia de Alimentos e pelo Programa de Pós-Graduação em Ciência e Tecnologia de Alimentos contou com a participação de palestrantes de renome internacional e nacional, além de profissionais atuantes nos seguimentos de regulamentação, aproveitamento e inserção de produtos regionais. Os participantes tiveram a oportunidade de debater e trocar experiências com investidores e empresários do setor e outros profissionais da área de alimentos em um ambiente onde a ciência e o setor econômico se encontraram, oportunizando a discussão de políticas que incentivam a transferência de tecnologias geradas nas instituições de pesquisa, transformando-as em produtos comerciais.

Microbial lipases are prominent biocatalysts able to catalyze a wide variety of reactions in aqueous and nonaqueous media. In this work, filamentous fungi isolated from leaves decomposed in an aquatic environment were screened for lipase production with hydrolytic activity and esterification. Agar plates with Tween 20 and Rhodamine B were used for selection, while submerged cultures with olive oil were subsequently used to select 38 filamentous fungi. Trichoderma harzianum, Fusarium solani, Trichoderma harzianum F5, and Penicillium sp. F36 were grown in six different culture media. F. solani presented the highest lipase production (2.37 U/mL) with esterification activity of 0.07 U/mL using medium composed of (g.L−1) KH2PO4 1.00, MgSO4 H2O 1.123, and CuSO4 0.06. Supplementation of this culture medium with organic nitrogen sources increased lipase production by 461.3% using tryptone and by 419.4% using yeast extract. Among the vegetable oils from the Amazon region, degummed cotton oil induced lipase production up to 8.14 U/mL. The lipase produced by F. solani F61 has great potential to application in conventional processes and biodiesel production by transesterification of vegetable oils, as well as food industries in the production of fatty acid esters by hydrolysis and esterification.

β-fructofuranosidase (invertase) andβ-D-fructosyltransferase (FTase) are enzymes used in industrial processes to hydrolyze sucrose aiming to produce inverted sugar syrup or fructooligosaccharides. In this work, a blackAspergillussp. PC-4 was selected among six filamentous fungi isolated from canned peach syrup which were initially screened for invertase production. Cultivations with pure carbon sources showed that invertase and FTase were produced from glucose and sucrose, but high levels were also obtained from raffinose and inulin. Pineapple crown was the best complex carbon source for invertase (6.71 U/mL after 3 days of cultivation) and FTase production (14.60 U/mL after 5 days of cultivation). Yeast extract and ammonium chloride nitrogen sources provided higher production of invertase (6.80 U/mL and 6.30 U/mL, respectively), whereas ammonium nitrate and soybean protein were the best nitrogen sources for FTase production (24.00 U/mL and 24.90 U/mL, respectively). Fermentation parameters for invertase using yeast extract wereYP/S= 536.85 U/g andPP= 1.49 U/g/h. FTase production showed values ofYP/S= 2,627.93 U/g andPP= 4.4 U/h using soybean protein. The screening for best culture conditions showed an increase of invertase production values by 5.10-fold after 96 h cultivation compared to initial experiments (fungi bioprospection), while FTase production increased by 14.60-fold (44.40 U/mL) after 168 h cultivation.A. carbonariusPC-4 is a new promising strain for invertase and FTase production from low cost carbon sources, whose synthesized enzymes are suitable for the production of inverted sugar, fructose syrups, and fructooligosaccharides.

Made available in DSpace on 2020-12-11T01:57:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-07-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Burkholderia glumae MA13 was isolated from soil samples of Atlantic rain forest ecosystem as a new bacterial strain for polyhydroxyalkanoate (PHA) production from crude glycerol as sole carbon source. Among 107 glycerol consuming bacterial isolates, B. glumae MA13 was cultivated in shake flask experiments in order to verify its PHA production capability from crude glycerol besides waste cooking oil and sugarcane molasses free of pretreatment, showing intracellular poly(3-hydroxybutyrate) [P(3HB)] accumulation values of 51.3, 51.4 and 49% of cell dry weight (CDW), respectively. The addition of propionic or hexanoic acids to the media containing crude glycerol resulted in the copolymers poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] comprised of up to 32.1 mol% of 3HV monomer or poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] with a maximum of 0.3 mol% of 3HHx, respectively. Fed-batch cultivation performed in a 10 L-scale bioreactor showed a maximum P(3HB-co-3HV) production of 9.9 g/L, which was 65.1% of total biomass and consisted of 21.8 mol% of 3HV monomer, with a maximum volumetric productivity of 0.22 g/(L h). The 3HV conversion yield achieved 1.04 g/g after 48 h cultivation, which was 76.7% of the maximum theoretical yield. B. glumae MA13 has showed to be an adapted bacterial strain to the synthesis of bioplastics from biofuel byproducts, and so from here it has been revealed as a promising PHA producer for an associated production set which has been considered as a prominent and ecologically friendly alternative to petrochemical plastics and fuels. Fed Univ Tocantins, Palmas Campus,Av NS 15,Quadra 109 Norte, BR-77001090 Palmas, TO, Brazil Sao Paulo State Univ, Inst Biosci, Av-24A,1515, BR-13506900 Rio Claro, SP, Brazil Sao Paulo State Univ, Bioenergy Res Inst, Av-24A,1515, BR-13506900 Rio Claro, SP, Brazil Fed Univ Tocantins, Gurupi Campus,Chacaras 69-72, BR-77920090 Gurupi, TO, Brazil Univ Sao Paulo, Inst Biomed Sci, Av Prof Lineu Prestes 1374, BR-05508900 Sao Paulo, SP, Brazil Sao Paulo State Univ, Inst Biosci, Av-24A,1515, BR-13506900 Rio Claro, SP, Brazil Sao Paulo State Univ, Bioenergy Res Inst, Av-24A,1515, BR-13506900 Rio Claro, SP, Brazil

The aim of this study was to evaluate eight plants extract for porcine pancreatic lipase inhibition, characterizing biochemically the extract with high inhibitory activity and its effects on preventing weight gain in female rat fed a high-fat diet (HFD). In vitro pancreatic lipase inhibition was carried out in ρ-nitrophenyl-laurate substrates and a double-reciprocal plot was used for inhibition mechanism identification. In vivo experiments, female rats were fed with a standard diet or high fat diet (HFD), HFD+orlistat, HFD+22.5 mg/ml GT, and HFD+112.5 mg/ml GT. Feed intake, weight body gain, fecal lipid excretion and biochemistry parameters were analyzed. Camellia sinensis extract had the highest inhibitory lipase activity (76.65 ± 2.04%) with a non-competitive inhibition. C. sinensis administration, equivalent to 112.5 mg/ml, promoted weight loss, while 22.5 mg/ml increased fecal excretion of lipids in 31.41%. C. sinensis extract is certainly a promising alternative for preventive obesity treatment, since biochemical parameters analyzed showed significantly, reduction in the serum triglycerides levels and significantly decreased the LDL-cholesterol basal levels when compared with animals that did not receive a fat diet. Key words: Enzyme inhibition, plant extracts, animal, preventive obesity.

Made available in DSpace on 2018-12-11T17:15:50Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-02-01 Microbial lipases are important enzymes in food and pharmaceutical industries. In this work, a Candida viswanathii lipase was purified by hydrophobic interaction chromatography on octyl Sepharose. The purification presented 78.4% yield and the enzyme was 8.7-fold purified, with specific activity 700.4 U/mg protein and 69 kDa molecular weight. Immobilization of the enzyme on the same support presented 72.5% yield and a derivative with 101% expressed activity (109.2 U/g support), indicating hyperactivation of the enzyme. Optimal activity for both free and immobilized lipase was observed at pH 4.0 and 45 °C. The free and immobilized lipase showed broad-range stability from acid to neutral pH, and apparent activation and stability on organic solvents. The derivative was 60-fold thermostabilized in relation to the free enzyme and fully retained its activity after four cycles of p-nitrophenyl palmitate hydrolysis. Slight activation was observed with dithiothreitol and β-mercaptoethanol. The free and immobilized lipase efficiently hydrolyzed monoesters, simple and mixed long chain triacylglycerols, as well as soy lecithin. The activity and stability in acid pH, the organic solvent tolerance and the lecithin hydrolysis indicate high potential application of the enzyme and its derivative in textile, food and pharma industries and for chemical synthesis. Bioprocess Engineering and Biotechnology Federal University of Tocantins – UFT, Rua Badejós, Chácaras 69/72, Zona Rural Biochemistry and Microbiology Department Bioscience Institute São Paulo State University IB/UNESP, Av. 24-A, 1515, Bela Vista Environmental Studies Center São Paulo State University CEA/UNESP, Av. 24-A, 1515, Bela Vista Biochemistry and Microbiology Department Bioscience Institute São Paulo State University IB/UNESP, Av. 24-A, 1515, Bela Vista Environmental Studies Center São Paulo State University CEA/UNESP, Av. 24-A, 1515, Bela Vista

Microorganisms can produce lipases with different biochemical characteristics making necessary the screening of new lipase-producing strains for different industrial applications. In this study, 90 microbial strains were screened as potential lipase producers using a sensitive agar plate method with a suitable medium supplemented with Tween 20 and also a liquid culture supplemented with olive oil. The highest cell growth and lipase production for Candida viswanathii were observed in triolein and oleic acid when used as the only pure carbon source. Renewable low-cost triacylglycerols supported the best cell growth, and olive oil was found to be the best inducer for lipase production (19.50 g/L and 58.50 U). The selected conditions for enzyme production were found with yeast extract as nitrogen source and 1.5 % (w/v) olive oil (85.70 U) that resulted in a good cell growth yield (YX/S = 1.234 g/g) and lipase productivity (1.204 U/h) after 72 h of shake-flask cultivation. C. viswanathii lipase presented high hydrolytic activity on esters bonds of triacylglycerols of long-chain, and this strain can be considered an important candidate for future applications in chemical industries.

Made available in DSpace on 2018-12-11T17:06:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01 The aims of this work were to establish improved conditions for lipase production by Candida viswanathii using agroindustrial wastes in solid-state cultivation and to purify and evaluate the application of this enzyme for poultry fat hydrolysis. Mixed wheat bran plus spent barley grain (1: 1, w/w) supplemented with 25.0% (w/w) olive oil increased the lipase production to 322.4%, compared to the initial conditions. When olive oil was replaced by poultry fat, the highest lipase production found at 40% (w/w) was 31.43 U/gds. By selecting, yeast extract supplementation (3.5%, w/w), cultivation temperature (30�C), and substrate moisture (40%, w/v), lipase production reached 157.33 U/gds. Lipase was purified by hydrophobic interaction chromatography, presenting a molecular weight of 18.5 kDa as determined by SDS-PAGE. The crude and purified enzyme showed optimum activity at pH 5.0 and 50�C and at pH 5.5 and 45�C, respectively. The estimated half-life at 50�C was of 23.5 h for crude lipase and 6.7 h at 40�C for purified lipase. Lipase presented high activity and stability in many organic solvents. Poultry fat hydrolysis was maximum at pH 4.0, reaching initial hydrolysis rate of 33.17 mmol/L/min. Thus, C. viswanathii lipase can be successfully produced by an economic and sustainable process and advantageously applied for poultry fat hydrolysis without an additional acidification step to recover the released fatty acids. Bioprocess Engineering and Biotechnology Federal University of Tocantins (UFT), Rua Badej�s, Ch�caras 69/72 Environmental Studies Center (CEA) Universidade Estadual Paulista (UNESP), Avenida 24-A, 1515 Bela Vista Biochemistry and Microbiology Department Bioscience Institute (IB) Universidade Estadual Paulista (UNESP), Avenida 24-A, 1515 Bela Vista Environmental Studies Center (CEA) Universidade Estadual Paulista (UNESP), Avenida 24-A, 1515 Bela Vista Biochemistry and Microbiology Department Bioscience Institute (IB) Universidade Estadual Paulista (UNESP), Avenida 24-A, 1515 Bela Vista

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:29:40Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:37:38Z : No. of bitstreams: 1 2-s2.0-84878682173.pdf: 1570551 bytes, checksum: aa05798f0135544fb1eeaaa44465d8b8 (MD5) Made available in DSpace on 2014-05-27T11:29:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-06-12 In recent decades, xylanases have been used in many processing industries. This study describes the xylanase production by Penicillium glabrum using brewer's spent grain as substrate. Additionally, this is the first work that reports the purification and characterization of a xylanase using this agroindustrial waste. Optimal production was obtained when P. glabrum was grown in liquid medium in pH 5.5, at 25 °C, under stationary condition for six days. The xylanase from P. glabrum was purified to homogeneity by a rapid and inexpensive procedure, using ammonium sulfate fractionation and molecular exclusion chromatography. SDS-PAGE analysis revealed one band with estimated molecular mass of 18.36 kDa. The optimum activity was observed at 60 °C, in pH 3.0. The enzyme was very stable at 50 °C, and high pH stability was verified from pH 2.5 to 5.0. The ion Mn2+ and the reducing agents β-mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg2+, Zn2+, and Cu2+ as well as the detergent SDS were strong inhibitors of the enzyme. The use of brewer's spent grain as substrate for xylanase production cannot only add value and decrease the amount of this waste but also reduce the xylanase production cost. © 2013 Adriana Knob et al. Department of Biological Sciences Midwest State University, Camargo Varela de Sá Street 03, 85040-080 Guarapuava, PR Department of Biology Midwest State University, Camargo Varela de Sá Street 03, 85040-080 Guarapuava, PR Department of Chemical Engineering Federal University of São Carlos Rodovia Washington Luís, km 235, SP-310, 13565-905 São Carlos, SP Department of Biochemistry and Microbiology São Paulo State University, 24-A Avenue 1515, 13506-900 Rio Claro, SP Department of Biochemistry and Microbiology São Paulo State University, 24-A Avenue 1515, 13506-900 Rio Claro, SP