Commercial and non-commercial pectinase and cellulase on the enzymatic hydrolysis efficacy of rice husk and Tifton 85 hay

The aim of this study was to evaluate the action of commercial and non-commercial cellulase and pectinase on rice husk and Tifton 85 hay hydrolyses. The hydrolysis kinetics of the substrates with commercial cellulase and pectinase were evaluated and the hydrolysis at different temperature and agitation conditions was maximized using experimental design. The combined use of commercial and non-commercial enzymes under optimized conditions was evaluated. The pre-treatment of the residues was also investigated by milling and different concentrations of NaOH. Finally, the effect of the hydrolysis on the bromatological composition of the residues was evaluated. The best hydrolysis times of rice husk and Tifton 85 hay were 10 and 12h for commercial cellulase, 12 and 14h for noncommercial cellulase, 10 and 14h for commercial pectinase and 16 and 20h for non-commercial pectinase, respectively. The highest hydrolysis values were obtained using commercial cellulase with 1:50 (w:v enzyme:water) dilution rate, at 45ºC and 300 rpm agitation for both substrates, reaching 20.6% maximum percentage for Tifton 85 hay and 11.6% for rice husk. The combined use of commercial enzymes did not increase hydrolysis percentage. The pre-treatment using 7.5% NaOH and 0.5 mm grain size significantly increased the rice husk and Tifton 85 hay hydrolyses (60-80%), either using commercial cellulase or pectinase enzymes. The use of non-commercial enzymes provided 18-30% hydrolysis obtained from commercial ones. Bromatological analyzes indicated a reduction in neutral detergent fiber and acid detergent fiber content for rice husk and Tifton 85 hay when using pectinases and commercial cellulases.(AU)

Commercial and non-commercial pectinase and cellulase on the enzymatic hydrolysis efficacy of rice husk and Tifton 85 hay

The aim of this study was to evaluate the action of commercial and non-commercial cellulase and pectinase on rice husk and Tifton 85 hay hydrolyses. The hydrolysis kinetics of the substrates with commercial cellulase and pectinase were evaluated and the hydrolysis at different temperature and agitation conditions was maximized using experimental design. The combined use of commercial and non-commercial enzymes under optimized conditions was evaluated. The pre-treatment of the residues was also investigated by milling and different concentrations of NaOH. Finally, the effect of the hydrolysis on the bromatological composition of the residues was evaluated. The best hydrolysis times of rice husk and Tifton 85 hay were 10 and 12h for commercial cellulase, 12 and 14h for noncommercial cellulase, 10 and 14h for commercial pectinase and 16 and 20h for non-commercial pectinase, respectively. The highest hydrolysis values were obtained using commercial cellulase with 1:50 (w:v enzyme:water) dilution rate, at 45ºC and 300 rpm agitation for both substrates, reaching 20.6% maximum percentage for Tifton 85 hay and 11.6% for rice husk. The combined use of commercial enzymes did not increase hydrolysis percentage. The pre-treatment using 7.5% NaOH and 0.5 mm grain size significantly increased the rice husk and Tifton 85 hay hydrolyses (60-80%), either using commercial cellulase or pectinase enzymes. The use of non-commercial enzymes provided 18-30% hydrolysis obtained from commercial ones. Bromatological analyzes indicated a reduction in neutral detergent fiber and acid detergent fiber content for rice husk and Tifton 85 hay when using pectinases and commercial cellulases.

Effect of dietary fiber concentrates on growth performance, gut morphology and hepatic metabolic intermediates in jundiá (Rhamdia quelen) / Efeito de concentrados de fibra alimentar sobre o desempenho de crescimento, morfologia intestinal e intermediários metabólicos hepáticos de jundiás (Rhamdia quelen)

A study was conducted to investigate the effect of Dietary Fiber Concentrates (DFCs) on growth performance, gut morphology, and hepatic metabolic intermediates in jundiá (Rhamdia quelen). At the end of the trial, growth and intestinal villus height was significantly (P< 0.05) higher in fish fed diets supplemented with DFCs. However, the animals in commercial prebiotic group showed higher values for this variable compared to the other treatments. Regarding the thickness of the epithelium bowel, it was greater in the Control group compared to animals supplemented with ß-glucan+mannan. Likewise, treatment with commercial prebiotic showed higher values of epithelium bowel compared to the DFCs. The fish supplemented with DFCs, had higher glycogen storage compared to the control group. These results indicate that DFCs can be considered as a beneficial dietary supplement for improving growth performance, gut morphology, and hepatic metabolic intermediates of jundiá.(AU)

O presente estudo foi conduzido para investigar o efeito de concentrados de fibras alimentares (CFAs) sobre o desempenho de crescimento, a morfologia intestinal e os parâmetros intermediários metabólicos hepáticos de jundiás (Rhamdia quelen). No final do experimento, o crescimento e a altura das vilosidades intestinais foram significativamente (P<0,05) maiores em peixes alimentados com dietas suplementadas com CFAs. No entanto, os animais suplementados com prebiótico comercial apresentaram valores mais elevados para essa variável em comparação com os outros tratamentos. Em relação à espessura do epitélio intestinal, esta foi maior nos animais do grupo controle em comparação com os animais suplementados com ß-glucano + manano. Da mesma forma, os peixes suplementados com prebiótico comercial apresentaram valores mais elevados do epitélio intestinal em comparação com os peixes suplementados com CFAs. Os peixes suplementados com CFAs obtiveram maior armazenamento de glicogênio em relação ao grupo controle. Esses resultados indicam que os CFAs podem ser utilizados como um suplemento alimentar benéfico para melhorar o desempenho do crescimento, a morfologia intestinal e os intermediários metabólicos hepáticos do jundiá.(AU)

Effect of dietary fiber concentrates on growth performance, gut morphology and hepatic metabolic intermediates in jundiá (Rhamdia quelen) / Efeito de concentrados de fibra alimentar sobre o desempenho de crescimento, morfologia intestinal e intermediários metabólicos hepáticos de jundiás (Rhamdia quelen)

A study was conducted to investigate the effect of Dietary Fiber Concentrates (DFCs) on growth performance, gut morphology, and hepatic metabolic intermediates in jundiá (Rhamdia quelen). At the end of the trial, growth and intestinal villus height was significantly (P< 0.05) higher in fish fed diets supplemented with DFCs. However, the animals in commercial prebiotic group showed higher values for this variable compared to the other treatments. Regarding the thickness of the epithelium bowel, it was greater in the Control group compared to animals supplemented with ß-glucan+mannan. Likewise, treatment with commercial prebiotic showed higher values of epithelium bowel compared to the DFCs. The fish supplemented with DFCs, had higher glycogen storage compared to the control group. These results indicate that DFCs can be considered as a beneficial dietary supplement for improving growth performance, gut morphology, and hepatic metabolic intermediates of jundiá.(AU)

O presente estudo foi conduzido para investigar o efeito de concentrados de fibras alimentares (CFAs) sobre o desempenho de crescimento, a morfologia intestinal e os parâmetros intermediários metabólicos hepáticos de jundiás (Rhamdia quelen). No final do experimento, o crescimento e a altura das vilosidades intestinais foram significativamente (P<0,05) maiores em peixes alimentados com dietas suplementadas com CFAs. No entanto, os animais suplementados com prebiótico comercial apresentaram valores mais elevados para essa variável em comparação com os outros tratamentos. Em relação à espessura do epitélio intestinal, esta foi maior nos animais do grupo controle em comparação com os animais suplementados com ß-glucano + manano. Da mesma forma, os peixes suplementados com prebiótico comercial apresentaram valores mais elevados do epitélio intestinal em comparação com os peixes suplementados com CFAs. Os peixes suplementados com CFAs obtiveram maior armazenamento de glicogênio em relação ao grupo controle. Esses resultados indicam que os CFAs podem ser utilizados como um suplemento alimentar benéfico para melhorar o desempenho do crescimento, a morfologia intestinal e os intermediários metabólicos hepáticos do jundiá.(AU)

Filamentous fungi producing enzymes under fermentation in cassava liquid waste / Fungos filamentosos produtores de enzimas sob fermentação em manipueira

The conversion of agroindustrial residues by microorganisms has been explored from fermentative processes to obtain several bioactive molecules. The objective of this work was to isolate and select filamentous fungi present in cassava liquid waste for the production of amylase, carboxymethylcellulose (CMCase), pectinase and xylanase using the same residue as induction substrate in fermentative processes. A total of 65 filamentous fungi were isolated and qualitative tests indicated that approximately 86% of these strains were able to produce at least one of the enzymes and 32% capable of producing the four enzymes. Fermentation assays in cassava liquid residue-containing medium showed 6 fungal lines as potential enzyme producers. The maximum activities of pectinase, xylanase, amylase and CMCase were respectively observed at 96 hours of fermentation by the strain by the strain Aspergillus sp. B5C; at 120 hours (163.6 ± 0.13 nKat mL-1), by Aspergillus sp. B4I; at 144 hours (99.8 ± 0.24 nKat mL-1), by Penicillium sp. B3A; and at 48 hours (55.5 ± 0.21 nKat mL-1), by Aspergillus sp. B4O. These results suggest that cassava liquid waste was source of filamentous fungi producing amylase, CMCase, pectinase and xylanase, as well as a promising alternative substrate for bioprocesses aiming the production of enzymes.

A conversão de resíduos agroindustriais por micro-organismos tem sido explorada a partir de processos fermentativos para obtenção de diversas moléculas bioativas. O objetivo deste trabalho foi isolar e selecionar fungos filamentosos presentes em manipueira para produção de amilase, carboximetilcelulase (CMCase), pectinase e xilanase utilizando o próprio resíduo como substrato indutor. Um total de 65 fungos filamentosos foi isolado e testes qualitativos indicaram que, aproximadamente, 86% dessas linhagens foram hábeis em produzir pelo menos uma das enzimas e 32% capazes de produzir as quatro enzimas. Ensaios fermentativos em meio contendo manipueira apontaram 6 linhagens fúngicas como potenciais produtoras de enzimas. As atividades máximas de pectinase, xilanase, amilase e CMCase foram observadas, respectivamente, às 96 horas de fermentação (67.4 ± 0,6 nKat mL-1), pela linhagem Aspergillus sp. B5C; às 120 horas (163.6 ± 0,13 nKat mL-1), por Aspergillus sp. B4I; às 144 horas (99.8 ± 0,24 nKat mL-1), por Penicillium sp. B3A; e às 48 horas (55.5 ± 0,21 nKat mL-1), por Aspergillus sp. B4O. Estes resultados sugerem a manipueira como fonte de fungos filamentosos produtores de amilase, CMCase, pectinase e xilanase, além de um promissor substrato alternativo para bioprocessos visando a produção dessas enzimas.

Filamentous fungi producing enzymes under fermentation in cassava liquid waste / Fungos filamentosos produtores de enzimas sob fermentação em manipueira

The conversion of agroindustrial residues by microorganisms has been explored from fermentative processes to obtain several bioactive molecules. The objective of this work was to isolate and select filamentous fungi present in cassava liquid waste for the production of amylase, carboxymethylcellulose (CMCase), pectinase and xylanase using the same residue as induction substrate in fermentative processes. A total of 65 filamentous fungi were isolated and qualitative tests indicated that approximately 86% of these strains were able to produce at least one of the enzymes and 32% capable of producing the four enzymes. Fermentation assays in cassava liquid residue-containing medium showed 6 fungal lines as potential enzyme producers. The maximum activities of pectinase, xylanase, amylase and CMCase were respectively observed at 96 hours of fermentation by the strain by the strain Aspergillus sp. B5C; at 120 hours (163.6 ± 0.13 nKat mL-1), by Aspergillus sp. B4I; at 144 hours (99.8 ± 0.24 nKat mL-1), by Penicillium sp. B3A; and at 48 hours (55.5 ± 0.21 nKat mL-1), by Aspergillus sp. B4O. These results suggest that cassava liquid waste was source of filamentous fungi producing amylase, CMCase, pectinase and xylanase, as well as a promising alternative substrate for bioprocesses aiming the production of enzymes.(AU)

A conversão de resíduos agroindustriais por micro-organismos tem sido explorada a partir de processos fermentativos para obtenção de diversas moléculas bioativas. O objetivo deste trabalho foi isolar e selecionar fungos filamentosos presentes em manipueira para produção de amilase, carboximetilcelulase (CMCase), pectinase e xilanase utilizando o próprio resíduo como substrato indutor. Um total de 65 fungos filamentosos foi isolado e testes qualitativos indicaram que, aproximadamente, 86% dessas linhagens foram hábeis em produzir pelo menos uma das enzimas e 32% capazes de produzir as quatro enzimas. Ensaios fermentativos em meio contendo manipueira apontaram 6 linhagens fúngicas como potenciais produtoras de enzimas. As atividades máximas de pectinase, xilanase, amilase e CMCase foram observadas, respectivamente, às 96 horas de fermentação (67.4 ± 0,6 nKat mL-1), pela linhagem Aspergillus sp. B5C; às 120 horas (163.6 ± 0,13 nKat mL-1), por Aspergillus sp. B4I; às 144 horas (99.8 ± 0,24 nKat mL-1), por Penicillium sp. B3A; e às 48 horas (55.5 ± 0,21 nKat mL-1), por Aspergillus sp. B4O. Estes resultados sugerem a manipueira como fonte de fungos filamentosos produtores de amilase, CMCase, pectinase e xilanase, além de um promissor substrato alternativo para bioprocessos visando a produção dessas enzimas.(AU)

Qualidade de melões Louis armazenados em quatro temperaturas / Quality of Louis melons stored under four storage temperatures

Este trabalho teve como objetivo estabelecer a melhor temperatura para a conservação de melões Louis. Os frutos foram armazenados sob condição de ambiente (22±2°C e 80±5% UR), sob refrigeração (3±1°C e 80±5% UR; 6±1°C e 80±5% UR; e 9±1°C e 80±5% UR) e avaliados quanto à coloração e firmeza da polpa, teores de sólidos solúveis, acidez titulável, atividade das enzimas peroxidase, polifenoloxidase, poligalacturonase e pectinametilesterase e perda de massa fresca. Os melões Louis armazenados a 22°C apresentaram maior redução nos teores de sólidos solúveis e acidez titulável, além de apresentarem maior atividade da enzima poligalacturonase, com conservação por até 18 dias. Os frutos armazenados a 6°C mantiveram a qualidade comercial durante 25 dias de armazenamento, apresentando os maiores teores de sólidos solúveis e menor atividade das enzimas peroxidase e poligalacturonase.(AU)

This research aimed to establish the best temperature for the conservation of Louis melons. Fruit were stored under ambient condition (22°C and 80% RH) and under refrigeration (3°C - 80% RH, 6°C - 90% RH e 9°C - 80% RH) and evaluated for color and resistance of pulp, soluble solids, titratable acidity content, peroxidase, polyphenol oxidase, polygalacturonase and pectin methyl esterase activity and weight loss. Louis melons at 22°C showed greater reduction in soluble solids and titratable acidity content, and had higher activity of the enzyme polygalacturonase, culminating in conservation for 18 days. Melons stored at 6°C preserved the shelf-life for 25 days, showed the highest soluble solids and lower activity of peroxidase and polygalacturonase.(AU)

Application of decolourized and partially purified polygalacturonase and alfa-amylase in apple juice clarification

Polygalacturonase and α-amylase play vital role in fruit juice industry. In the present study, polygalacturonase was produced by Aspergillus awamori Nakazawa MTCC 6652 utilizing apple pomace and mosambi orange (Citrus sinensis var mosambi) peels as solid substrate whereas, α-amylase was produced from A. oryzae (IFO-30103) using wheat bran by solid state fermentation (SSF) process. These carbohydrases were decolourized and purified 8.6-fold, 34.8-fold and 3.5-fold, respectively by activated charcoal powder in a single step with 65.1%, 69.8% and 60% recoveries, respectively. Apple juice was clarified by these decolourized and partially purified enzymes. In presence of 1% polygalacturonase from mosambi peels (9.87 U/mL) and 0.4% α-amylase (899 U/mL), maximum clarity (%T660nm = 97.0%) of juice was attained after 2 h of incubation at 50 ºC in presence of 10 mM CaCl2. Total phenolic content of juice was reduced by 19.8% after clarification, yet with slightly higher %DPPH radical scavenging property.(AU)

Caracterização física, química e bioquímica do mesocarpo interno de frutos do pequizeiro colhidos em diferentes estádios de desenvolvimento

The aim of this study was to characterize physical, chemical and biochemical pequi fruit produced in southern Minas Gerais, Brazil, during their development stages. Fruits collected in the 8th, 9th, 10th, 12th and 13th weeks after anthesis, were selected for size, skin color, and absence of injuries and evaluated according to firmness, levels of beta-carotene, total soluble sugars, and total and soluble pectins as well as the activity of pectinmethylesterase (PME), polygalacturonase (PG), peroxidase (POD) and polyphenoloxidase (PPO). It was found that with the advancement of days after anthesis occurs in pequi fruits the synthesis of beta-carotene and total soluble sugars, reducing firmness, pectic solubilization, absence of PME activity, decrease in PG activity and increased in activities POD and PPO in the pulp.

O objetivo deste trabalho foi a caracterização física, química e bioquímica do pequi, produzido no sul de Minas Gerais, ao longo do seu desenvolvimento. Frutos coletados na 8ª, 9ª, 10ª, 12ª e 13ª semanas após a antese foram selecionados quanto ao tamanho, cor da casca, ausência de injúrias e avaliados quanto à firmeza da polpa, teores de beta-caroteno, de açúcares solúveis totais e de pectinas total e solúvel, assim como quanto à atividade das enzimas pectinametilesterase (PME), poligalacturonase (PG), peroxidase (POD) e polifenoloxidase (PPO). Verificou-se que, com o avanço dos dias, após a antese nos frutos do pequizeiro, ocorre a síntese do beta-caroteno e de açúcares solúveis totais, redução da firmeza, solubilização das pectinas, ausência de PME, diminuição na atividade da PG e aumento nas atividades da POD e da PPO na polpa.

Caracterização física, química e bioquímica do mesocarpo interno de frutos do pequizeiro colhidos em diferentes estádios de desenvolvimento

The aim of this study was to characterize physical, chemical and biochemical pequi fruit produced in southern Minas Gerais, Brazil, during their development stages. Fruits collected in the 8th, 9th, 10th, 12th and 13th weeks after anthesis, were selected for size, skin color, and absence of injuries and evaluated according to firmness, levels of beta-carotene, total soluble sugars, and total and soluble pectins as well as the activity of pectinmethylesterase (PME), polygalacturonase (PG), peroxidase (POD) and polyphenoloxidase (PPO). It was found that with the advancement of days after anthesis occurs in pequi fruits the synthesis of beta-carotene and total soluble sugars, reducing firmness, pectic solubilization, absence of PME activity, decrease in PG activity and increased in activities POD and PPO in the pulp.

O objetivo deste trabalho foi a caracterização física, química e bioquímica do pequi, produzido no sul de Minas Gerais, ao longo do seu desenvolvimento. Frutos coletados na 8ª, 9ª, 10ª, 12ª e 13ª semanas após a antese foram selecionados quanto ao tamanho, cor da casca, ausência de injúrias e avaliados quanto à firmeza da polpa, teores de beta-caroteno, de açúcares solúveis totais e de pectinas total e solúvel, assim como quanto à atividade das enzimas pectinametilesterase (PME), poligalacturonase (PG), peroxidase (POD) e polifenoloxidase (PPO). Verificou-se que, com o avanço dos dias, após a antese nos frutos do pequizeiro, ocorre a síntese do beta-caroteno e de açúcares solúveis totais, redução da firmeza, solubilização das pectinas, ausência de PME, diminuição na atividade da PG e aumento nas atividades da POD e da PPO na polpa.

Screening and optimization of pectin lyase and polygalacturonase activity from ginseng pathogen Cylindrocarpon Destructans

Cylindrocarpon destructans isolated from ginseng field was found to produce pectinolytic enzymes. A Taguchi's orthogonal array experimental design was applied to optimize the preliminary production of polygalacturonase (PG) and pectin lyase (PL) using submerged culture condition. This method was applied to evaluate the significant parameters for the production of enzymes. The process variables were pH, pectin concentration, incubation time and temperature. Optimization of process parameters resulted in high levels of enzyme (PG and PL) production after ten days of incubation at a pH of 5.0 at 25°C in the presence of 1.5% pectin. Among different nitrogen sources, urea and peptone showed high production of PG and PL, respectively. The enzyme production and mycelial growth seems to have direct influence on the culture conditions; therefore, at stationary state high enzyme production and mycelial growth were obtained than agitation state. Along with this, optimization of enzyme activity was also determined using various physiological parameters like, temperature, incubation time and pH. Taguchi's data was also analyzed using one step ANOVA statistical method.

Xylanase production by Aspergillus awamori under solid state fermentation conditions on tomato pomace

In this work, tomato pomace, a waste abundantly available in the Mediterranean and other temperate climates agro-food industries, has been used as raw material for the production of some hydrolytic enzymes, including xylanase, exo-polygalacturonase (exo-PG), cellulase (CMCase) and -amylase. The principal step of the process is the solid state fermentation (SSF) of this residue by Aspergillus awamori. In several laboratory experiments, maximum xylanase and exo-PG activities were measured during the first days of culture, reaching values around 100 and 80 IU/gds (international units of enzyme activity per gram of dried solid), respectively. For CMCase and -amylase production remained almost constant along fermentation, with average values of 19 and 21.5 IU/gds, respectively. Experiments carried out in a plate-type bioreactor at lab scale showed a clear positive effect of aeration on xylanase and CMCase, while the opposite was observed for exo-PG and -amylase. In general, xylanase was the enzyme produced in higher levels, thus the optimum conditions for the determination of the enzyme activity was characterized. The xylanase activity shows an optimum pH of 5 and an optimum temperature of 50 ºC. The enzyme is activated by Mg2+, but strongly inhibited by Hg2+ and Cu2+. The enzymatic activity remains quite high if the extract is preserved in a range of pH from 3 to 10 and a temperature between 30 ºC to 40 ºC.

Exo-polygalacturonase production by Bacillus subtilis CM5 in solid state fermentation using cassava bagasse

The purpose of this investigation was to study the effect of Bacillus subtilis CM5 in solid state fermentation using cassava bagasse for production of Exo-polygalacturonase (exo-PG). Response surface methodology was used to evaluate the effect of four main variables, i.e. incubation period, initial medium pH, moisture holding capacity (MHC) and incubation temperature on enzyme production. A full factorial Central Composite Design was applied to study these main factors that affected exo-PG production. The experimental results showed that the optimum incubation period, pH, MHC and temperature were 6 days, 7.0, 70% and 50ºC, respectively for optimum exo-PG production.

O objetivo desta investigação foi estudar a produção de exo-poligalacturonase (exo-PG) por Bacillus subtilis CM5 por fermentação em estado sólido empregando bagaço de mandioca. Empregou-se a metodologia de superfície de resposta para avaliar o efeito de quatro variáveis na produção da enzima: período de incubação, pH inicial do meio, MHC e temperatura de incubação. Os resultados experimentais mostraram que os ótimos de temperatura, período de incubação, MHC e temperatura para produção de exo-PG foram seis dias, 7,0, 70% e 50ºC, respectivamente.

Novel process for the simultaneous extraction and degumming of banana fibers under solid-state cultivation

Various process parameters for the production of polygalacturonase by Streptomyces lydicus under solid-state fermentation were optimized. The optimum particle size of wheat bran for polygalacturonase production was in the range of 500-1000 µm. Initial moisture content of 70% was found to be the optimum for enzyme production. The most suitable inoculum size was 1.25 x 10(5) CFU/mL and the optimum incubation temperature was 30ºC. Addition of carbon sources resulted in 37% increase in enzyme yield (425 U/g), whereas no significant enhancement was obtained on nitrogen supplementation. Maximum enzyme yield was recorded at 72 h. When compared to the initial production medium (108.5 U/g), the enzyme yield was 3.9 fold after optimization. Solid-state fermentation was effectively employed to develop a novel process for the simultaneous extraction and degumming of banana fibers. Streptomyces lydicus was allowed to grow on wheat bran medium in which banana leaf sheath pieces were incorporated and the fiber bundles were separated after a two-step fermentative process.

Vários parâmetros de processo de produção de poligalacturonase por Streptomyces lydicus por fermentação em estado sólido foram otimizados. O tamanho ótimo de partícula de farelo de trigo para a produção de poligalacturonase esteve na faixa de 500 a 1000 mm. O teor inicial de umidade de 70% foi o melhor para a produção da enzima. O inóculo inicial mais adequado foi de 1,25 x 10(5) UFC/mL e a temperatura ótima de incubação foi 30ºC. A adição de fontes de carbono resultou em aumento de 37% no rendimento da enzima (425U/g), enquanto que a suplementação com nitrogênio não melhorou o rendimento. O rendimento máximo da enzima foi obtido em 72h. A otimização resultou em um aumento de 3,9 vezes na quantidade de enzima produzida inicialmente (108,5U/g). A fermentação em estado-sólido foi eficiente para o desenvolvimento de um novo processo de extração e simultânea degomagem. Streptomyces lydicus foi cultivado em meio de farelo de trigo acrescentado de fragmentos de folhas de banana, sendo os feixes de fibras separados após um processo de fermentação em dois passos.

Isolation of recombinant strains with enhanced pectinase production by protoplast fusion between Penicillium expansum and Penicillium griseoroseum

Protoplast fusion between complementary auxotrophic and morphological mutant strains of Penicillium griseoroseum and P. expansum was induced by polyethylene glycol and calcium ions (Ca2+). Fusant strains were obtained in minimal medium and a prototrophic strain, possibly diploid, was chosen for haplodization with the fungicide benomyl. Different recombinant strains were isolated and characterized for occurrence of auxotrophic mutations and pectinolytic enzyme production. The fusant prototrophic did not present higher pectinase production than the parental strains, but among 29 recombinants analyzed, four presented enhanced enzyme activities. The recombinant RGE27, which possesses the same auxotrophic and morphologic mutations as the P. griseoroseum parental strain, presented a considerable increase in polygalacturonase (3-fold) and pectin lyase production (1.2-fold).

Fusões de protoplastos entre linhagens mutantes auxotróficas e morfológicas complementares de Penicillium griseoroseum e P. expansum foram induzidas por polietilenoglicol e íons cálcio (Ca2+). Fusionantes foram obtidos em meio mínimo e uma linhagem prototrófica, possivelmente diplóide, foi selecionada para a haploidização com o fungicida benomil. Diferentes linhagens recombinantes foram isoladas e caracterizadas quanto à presença de mutações auxotróficas e a produção de enzimas pectinolíticas. O fusionante prototrófico não apresentou maior atividade de pectinases em relação às linhagens parentais, entretanto, entre 29 recombinantes analisados, quatro apresentaram maiores atividades enzimáticas. O recombinante RGE27, o qual possui as mesmas mutações auxotróficas e morfológicas que a linhagem parental de P. griseoroseum, apresentou um aumento considerável na produção de poligalacturonase (3 vezes) e de pectina liase (1,2 vezes).

Column bioreactor use for optimization of pectinase production in solid substrate cultivation

This study aimed to determine the influence of process variables and production, of polygalacturonase (PG) and polymetylgalacturonase (PMG) by solid substrate cultivation using a fixed bed column bioreactor. A fractional factorial design (FFD) was used to study the effect of the following variables: microorganism (Aspergillus oryzae and Aspergillus niger), substratum (wheat bran and defatted rice bran), aeration (40 and 60 ml h-1g-1), pectin (5 and 10 g g-1) and nitrogen (urea and ammonium sulfate). Microorganism, aeration and initial pectin were identified in FFD as significant variables (p 0.05) on PG production. A central composed design to optimize PG and PMG productions indicated that Aspergillus niger presents higher PG production; substratum and nitrogen do not affect PG production; the aeration rate affects positively the production of PG and negatively the production of PMG and the initial pectin concentration affects positively both PG and PMG production. The optimal point of aeration and initial pectin concentration for PG production are 66.13 ml h-1 g-1 and 12.8 g g-1, respectively and for PMG production are 40 ml h-1 g-1 and 15.0 g g-1, respectively.

O objetivo deste trabalho foi determinar a influência de variáveis de processo na produção das enzimas poligalacturonase (PG) e polimetilgalacturonase (PMG) por cultivo semi-sólido utilizando bioreator de coluna. Um planejamento fatorial fracionário foi utilizado para determinar o efeito das variáveis "microrganismo" (Aspergillus oryzae e Aspergillus niger), "substrato" (farelo de trigo e farelo desengordurado de arroz), "aeração" (40 e 60 ml h-1 g-1), "pectina" (5 e 10 g g-1) e "nitrogênio" (uréia e sulfato de amônia) para a produção de PG. Microrganismo, aeração e pectina foram significantes (p 0,05) para a produção de PG. Utilizando-se um planejamento composto central com quadruplicata no ponto central concluiu-se que Aspergillus niger apresenta maior produção de PG, o substrato e a fonte de nitrogênio não afetam (p 0,05) a produção de PG, a aeração afeta positivamente a produção de PG e negativamente a de PMG, e a concentração inicial de pectina afeta positivamente a atividade de ambas enzimas. Os pontos ótimos de aeração e de concentração inicial de pectina para a produção de PG são 66,13 ml h-1 g-1 e 12,8 g g-1, respectivamente, e para a produção de PMG são 40 ml h-1 g-1 e 15,0 g g-1, respectivamente.

Production and partial characterization of polygalacturonases produced by thermophilic Monascus sp N8 and by thermotolerant Aspergillus sp N12 on solid-state fermentation

Polygalacturonases production by newly isolated Monascus sp N8 and Aspergillus sp N12 strains was carried out in solid-state fermentation using mixtures of wheat bran, sugar cane bagasse and orange bagasse as carbon sources. The maximal activity values of exo-polygalacturonases (exo-Pg) from Monascus sp and Aspergillus sp were obtained using wheat bran/sugar cane bagasse/orange bagasse mixture (6.6 U/mL) and wheat bran/orange bagasse mixture (10 U/mL), respectively. Enzyme production by both strains was higher at 45ºC after 72 h and 1.6 U/mL at 50ºC after 120 h. Endo-polygalacturonase (endo-Pg) production was higher in wheat bran/orange bagasse mixture and was not affected by temperature of incubation for both fungi. Endo-Pg production by Monascus was 1.8 U/mL at 45ºC and 50ºC, after 72. Similar values were obtained in Aspergillus sp culture, 1.9 U/mL at 45ºC and 1.8 U/mL at 50ºC. Exo-Pg from both strains showed optimum activity at pH 5.5. Maximal activity was determined at 60ºC for enzyme from Monascus sp and 50ºC for that produced by Aspergillus sp. Exo-Pg from Monascus sp was stable at pH range 4.5-6.0 whereas that from Aspergillus sp enzyme was stable at pH 4.0. Both enzymes showed stability when incubated at 50ºC for 1 h, in absence of substrate.

A produção de poligalacturonases pelas linhagens fúngicas recentemente isoladas, Monascus sp N8 e Aspergillus sp N12, foi estudada através de fermentação em estado sólido usando como substratos misturas de farelo de trigo, bagaço da cana-de-açúcar e bagaço de laranja. A atividade máxima de exo-Pg produzida por Monascus sp (6,6 U/mL) foi obtida quando o meio de cultivo utilizado continha mistura de farelo de trigo, bagaço da cana-de-açúcar e bagaço de laranja (1:1:1), enquanto que Aspergillus sp produziu maior quantidade da enzima (10 U/mL) em meio de farelo de trigo e bagaço de laranja. A maior produção de exo-Pg foi obtida através de incubação das culturas a 45ºC quando comparadas àquelas incubadas a 50ºC. A produção de endo-poligalacturonase (endo-Pg) pelas duas linhagens não foi afetada pela temperatura de incubação. A atividade de endo-Pg em meio de cultura Monascus sp foi 1.8 U/mL a 45ºC em 72 hs de fermentação e 1,6 U/mL a 50ºC em 120 hs de fermentação nas mesmas condições. Valores semelhantes foram obtidos pelo cultivo de Aspergillus sp com 1.9 U/mL a 45ºC a 1.8 U/mL at 50ºC. As exo-poligalacturonases produzidas por ambas as linhagens mostraram maiores atividades em pH 5,5. Enzimas de Monascus sp foi mais ativa a 60ºC e a de Aspergillus sp, a 50ºC. A exo-Pg produzida por Monascus sp foi estável em valores de pH entre 4,5-6,0, enquanto a de Aspergillus sp foi estável somente em pH 4,0. Ambas as enzimas mostraram-se estáveis por 1 hora a 50ºC, quando incubadas em ausência de substrato.

Pectinase and polygalacturonase production by a thermophilic Aspergillus fumigatus isolated from decomposting orange peels

A thermophilic fungal strain producing both pectinase and polygalacturonase was isolated after primary screening of 120 different isolates. The fungus was identified as Aspergillus fumigatus Fres. MTCC 4163. Using solid-state cultivation, the optimum levels of variables for pectinase and polygalacturonase (PG) production were determined. Maximal levels of enzyme activities were achieved upon growing the culture in a medium containing wheat bran, sucrose, yeast extract and (NH4)2SO4 after 2-3 days of incubation at a temperature of 50ºC. Highest enzyme activities of 1116 Ug-1 for pectinase and 1270 Ug-1 for polygalacturonase were obtained at pH 4.0 and 5.0, respectively.

Através da tiragem de 120 cepas de fungos, isolou-se uma cepa capaz de produzir tanto pectinase quanto poligalacturonase. A cepa foi identificada como Aspergillus fumigatus Fres. MTCC 4163. Empregando cultivo em estado sólido, determinou-se os níveis ótimos das variáveis para a produção de pectinase e de poligalacturonase. Os níveis máximos de atividade enzimática foram obtidos quando a cultura era realizada em meio contendo farelo de trigo, sacarose, extrato de levedura e (NH4)2SO4 por 2-3 dias a uma temperatura de 50ºC. A atividade máxima de pectinase (1116 Ug-1) e de poligalacturonase (1270 Ug-1) foi obtida em pH 4,0 e 5,0, respectivamente.

Suculência e solubilização de pectinas em maçãs ‘Gala, armazenadas em atmosfera controlada, em dois níveis de umidade relativa

The objective of this study was to evaluate the effect of relative humidities (RH) (96 and 90%), in apples ‘Gala stored by 8 months in controlled atmosphere (CA) on the juiciness loss, flesh firmness, soluble pectin content (SPC) and activities of pectinmethylesterase (PME) and polygalacturonase (PG). The storage temperature was 0.5°C. At the end of the CA period and seven more days at 20°C were evaluated the juiciness, flesh firmness, SPC, PME and PG. RH of 96 to 90%, did not affect the juiciness, firmness, SPC and PME activity, but 90% of RH increased PG activity, after 8 months in AC and 7 more days at 20°C.

O objetivo deste trabalho foi o de avaliar o efeito de umidades relativas (UR) de 96 e 90%, em maçãs ‘Gala armazenadas por 8 meses em atmosfera controlada (AC) sobre a perda de suculência, firmeza de polpa, conteúdo de pectina solúvel (PS) e atividade das enzimas pectinametilesterase (PME) e poligalacturonase (PG). A temperatura de armazenamento em AC foi de 0,5°C. Ao final do período em AC e mais 7 dias a 20°C, foram avaliadas a suculência, a firmeza de polpa, o conteúdo de PS e as atividades da PME e PG. A UR na faixa de 96 a 90% não afetou a suculência, a firmeza, o conteúdo de PS e a atividade da PME, mas em 90% de UR ocorreu aumento da atividade da PG, após 8 meses em AC e mais 7 dias a 20°C.

Suculência e solubilização de pectinas em maçãs ‘Gala’, armazenadas em atmosfera controlada, em dois níveis de umidade relativa

The objective of this study was to evaluate the effect of relative humidities (RH) (96 and 90%), in apples ‘Gala’ stored by 8 months in controlled atmosphere (CA) on the juiciness loss, flesh firmness, soluble pectin content (SPC) and activities of pectinmethylesterase (PME) and polygalacturonase (PG). The storage temperature was 0.5°C. At the end of the CA period and seven more days at 20°C were evaluated the juiciness, flesh firmness, SPC, PME and PG. RH of 96 to 90%, did not affect the juiciness, firmness, SPC and PME activity, but 90% of RH increased PG activity, after 8 months in AC and 7 more days at 20°C.

O objetivo deste trabalho foi o de avaliar o efeito de umidades relativas (UR) de 96 e 90%, em maçãs ‘Gala’ armazenadas por 8 meses em atmosfera controlada (AC) sobre a perda de suculência, firmeza de polpa, conteúdo de pectina solúvel (PS) e atividade das enzimas pectinametilesterase (PME) e poligalacturonase (PG). A temperatura de armazenamento em AC foi de 0,5°C. Ao final do período em AC e mais 7 dias a 20°C, foram avaliadas a suculência, a firmeza de polpa, o conteúdo de PS e as atividades da PME e PG. A UR na faixa de 96 a 90% não afetou a suculência, a firmeza, o conteúdo de PS e a atividade da PME, mas em 90% de UR ocorreu aumento da atividade da PG, após 8 meses em AC e mais 7 dias a 20°C.