Lipase Production by Aspergillus niger C by Submerged Fermentation

Abstract This study aimed to evaluate the effects of variables on the process of lipases production by Aspergillus niger C by submerged fermentation (SmF). The production assays were performed in shake flasks for 72 hours at 150 rpm and 32°C. First, a fractional factorial design 25-1 (FFD) was carried out to evaluate the effect of the following process variables: sucrose, ammonium sulphate, soybean oil, yeast extract concentration and pH. After the selection of the variables that significantly influenced the lipase production, a central composite rotational design 22 (CCRD) was used, aiming to find the most favorable operational conditions. The selected assay condition (15.0 g.L-1 sucrose, 4.0 g.L-1 ammonium sulphate, 4.0 g.L-1 soybean oil and 1.0 g.L-1 yeast extract at pH 5.0) was the one that presented a lipase activity of 27.46 U.mL-1. It was very close to that best assay (30.76 U.mL-1), but using half of the inducer concentration, consequently reducing process cost. The kinetics of lipase production showed that the highest specific activity was 57.17 U.mg-1. The pH and temperature effects on lipase activity produced in this study was investigated. The optimum activity was found in a more acidic pH (5.0-6.0) and 55°C.

Cassava Pulp Enzymatic Hydrolysate as a Promising Feedstock for Ethanol Production

ABSTRACT The aim of this study was to produce bioethanol from enzymatic hydrolysates of cassava pulp, a by-product of cassava flour manufacturing, using an alcohol-tolerant Saccharomyces cerevisiae strain. First, the best operational condition of the starch hydrolysis process was determined through a complete factorial design (24), with triplicates at the central point. The independent variables evaluated were: the concentrations of α-amylase (Termamyl 2X) and glucoamylase (AMG 300L) and both liquefaction and saccharification times. The most favorable hydrolysis condition in the assay was achieved using 0.517 mL AMG.g starch-1 and 0.270 mL Termamyl.g starch-1, with liquefaction and saccharification times of 1 and 2 h, respectively. The broth obtained at this hydrolysis condition contained a high glucose concentration (160 g.L-1). Once the best reaction conditions were determined, fermentation tests were carried out in a 3 L bioreactor, in a batch system, at 30 °C, 100 rpm and pH 5.5, using 3 g.L-1 (dry biomass) of yeast as inoculum. After 24 h of fermentation, an ethanol concentration of 68 g.L-1 was obtained, with 0.48 ethanol yield and 2.83 g.L-1.h-1 productivity. These results indicate the potential use of cassava pulp, a by-product of cassava flour industries in Brazil, as a raw material for bioethanol production.

Selection of cellulolytic fungi isolated from diverse substrates

The aim of the present work was to select filamentous fungi isolated from diverse substrates to obtain the strains with potential to produce the hydrolytic enzymes. From a total of 215 strains, seven strains from the soils, six from the plants and one from sugarcane bagasse were selected and identified as belonging to the Trichoderma, Penicillium and Aspergillus genera. The best hydrolytic activities obtained by semi-solid fermentation using these strains were approximately: 35; 1; 160; 170 and 120 U/gdm (CMCase, FPase, β-glucosidase, xylanase and polygalacturonase, respectively), demonstrating their potential to synthesize the enzymes compared with the results reported in the literature.

Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9

An extracellular phytase from Aspergillus niger 11T53A9 was purified about 51-fold to apparent homogeneity with a recovery of 20.3 percent referred to the phytase activity in the crude extract. Purification was achieved by ammonium sulphate precipitation, ion chromataography and gel filtration. The purified enzyme behaved as a monomeric protein with a molecular mass of about 85 kDa and exhibited maximal phytate-degrading activity at pH 5.0. Optimum temperature for the degradation of phytate was 55ºC. The kinetic parameters for the hydrolysis of sodium phytate were determined to be K M = 54 µmol l-1 and k cat = 190 sec-1 at pH 5.0 and 37ºC. The purified enzyme was rather specific for phytate dephosphorylation. It was shown that the phytase preferably dephosphorylates myo-inositol hexakisphosphate in a stereospecific way by sequential removal of phosphate groups via D-Ins(1,2,4,5,6)P5, D-Ins(1,2,5,6)P4, D-Ins(1,2,6)P3, D-Ins(1,2)P2 to finally Ins(2)P.

Utilization of agroindustrial residues for lipase production by solid-state fermentation / Utilização de resíduos agroindustriais para produção de lipase por fermentação em estado sólido

The aim of this work was to produce lipases by solid-state fermentation (SSF) using, as substrate, agroindustrial residue supplemented with by-products from corn oil refining process or olive oil. For a group of ten fungi strains selected in the first steps, the lipase activity obtained by SSF varied from 7.7 to 58.6 U/g of dry substrate (gds). Among the evaluated strains, the Aspergillus niger mutant 11T53A14 was selected by presenting the best enzymatic production. For the fermentation tests, two substrates were also investigated: wheat bran and corn cob, both supplemented with olive oil. The best results were obtained with wheat bran. Additionally, three industrial by-products from corn oil refining (soapstock, stearin and fatty acids) were evaluated as substitutes to the olive oil in the function of lipases production inducer. Among them, soapstock and stearin were the best inducers, whereas fatty acids presented an inhibitor effect. The highest lipase activities using soapstock, stearin and fatty acids were 62.7 U/gds, 37.7 U/gds and 4.1 U/gds, respectively.

O objetivo deste trabalho foi produzir lipases por fermentação em estado sólido (FES) utilizando, como substrato, resíduo agroindustrial enriquecido com subprodutos do processo de refino do óleo de milho ou óleo de oliva. Para um conjunto de dez linhagens de fungos selecionadas nas primeiras etapas, a atividade lipásica obtida por FES variou de 7,7 a 58,6 U/g de substrato seco (gss). Dentre as linhagens avaliadas, o mutante Aspergillusniger 11T53A14 foi selecionado por apresentar a melhor produção enzimática.Para os testes de fermentação, dois substratos foram investigados: farelo de trigo e sabugo de milho, ambos enriquecidos com óleo de oliva. Nestes testes, os melhores resultados foram obtidos com farelo de trigo. Adicionalmente, três subprodutos industriais do refino do óleo de milho (borra, estearina e ácidos graxos) foram avaliados como substitutos do óleo de oliva na função de indutor para a produção de lipases. Dentre eles, borra e estearina demonstraram ser melhores indutores, enquanto ácidos graxos apresentaram um efeito inibidor. As mais altas atividades lipásicas utilizando borra, estearina e ácidos graxos foram 62,7 U/gss, 37,7 U/gss e 4,1 U/gss, respectivamente.

Tanase: conceitos, produção e aplicação / Tannase: concepts, production and application

O objetivo deste trabalho foi efetuar revisão do conhecimento sobre tanase, apontando necessidades e perspectivas de utilização, em especial para a área de alimentos. Foram enfatizados tópicos como importância fisiológica da tanase e seu mecanismo de ação, sua produção por microorganismos, os métodos analíticos para sua determinação, a sua produção por fermentãção, a regulação da biosíntese, suas características e propriedades, a imobilização e suas aplicações. A principal aplicação dessa enzima encontra-se na produção de chás instantâneos, porém outros campos potenciais podem ser vislumbrados como sua utilização para elaboração de rações animais com maior valor nutricional, produção de compostos antioxidantes e produçaõ de ácido gálico

Influence of metal ions on pellet morphology and polygalacturonase synthesis by Aspergillus niger 3T5B8

The effects of cations addition on pellet morphology and polygalacturonase (PG) synthesis by Aspergillus niger 3T5B8 were studied and compared with a control system. Fe(II), Cu(II), Zn(II) and Mn(II) were added to the fermentation medium separately, and also as combined groups of cations. The addition of Fe2+ and/or Zn2+ ions was significantly positive to the enzyme production. A positive effect in the biomass content, however, was only obtained when the same metal ions were added separately. On the other hand, Cu2+ and Mn2+ ions had almost no effect on these parameters. The morphology of the pellets was studied by image processing techniques. Small pellets with small cores were usually obtained when Fe2+ and Zn2+ ions were individually or collectively added to the medium. The pellets produced in media containing Fe2+ or Zn2+ ions were compact, while the ones produced in a medium containing both cations were considered diffuse. Autolysis of the core was observed for large control pellets, due to the deficient nutrient transfer to the interior of the pellet. The pellets obtained in a medium containing both Fe2+ and Zn2+ ions were high enzyme producers, probably due to a loose morphology, induced by the presence of combined groups of metal ions in the medium, favoring the nutrient transfer.

Comparaçäo entre os tratamentos com tanase e com gelatina para clarificaçäo do suco de caju (Anacardium occidentale L) / Comparison of the cashew apple (Anacardium occidentale L) juice clarification with tannase and gelatin

Comparou-se o tratamento da polpa do caju com tanase e com gelatina, visando à reduçäo da turbidez e do teor de taninos. Os resultados indicaram que a reduçäo das concentraçöes de taninos totais, hidroslisáveis, proantocianidinas e turbidez ficaram em 30 por cento, 50 por cento, 32 por cento e 94 por cento para sucos tratados com gelatina e 46 por cento 88 port cento e 2 por cento e 88 por cento para a bolpa tratada com tanase, respectivamente. Deste modo, o tratamento com gelatina foi eficiente para remoçäo de proantocianidinas, enquanto que o tratamento enzimático foi mais eficiente na reduçäo de taninos hidrolisáveis. Os produtos obtidos näo apresentaram diferenças vissuais. Estudo comparativo de pré-vialidadeeconömica mostrou que o investimento de capital para obtençäo do suco de caju clarificado com enzima é menor que para o suco clarificado com gelatina. Entretanto, o custo de produçäo do suco de caju clarificado com gelatina é menor devido ao preço elevado da enzima tanase

Selection of tannase-producing Aspergillus niger strains

The aim of this work was to select strains of Aspergillus niger for tannase production. Growth of colonies in plates with tannic acid-containing medium indicated their ability to synthesize tannase. Tannase activity was also measured in solid-state fermentation. A. niger 11T25A5 was the best tannase producer (67.5 U.g-1/72 hours of fermentation.

Genetic manipulation of Aspergillus niger for increased synthesis of pectinolytic enzymes

Um grupo de 35 mutantes morfológicos com potencial para síntese de enzimas pectinolíticas foi obtido de conídios de A. niger, empregando a técnica convencional de mutaçäo induzida por EMS seguida por irradiaçäo com UV. A linhagem mutante selecionada na primeira etapa (descrita como A. niger 5T25A) foi usada para a segunda etapa de induçäo de mutaçäo. Todas as linhagens selecionadas foram testadas para a síntese de enzimas pectinolíticas por fermentaçäo em estado sólido. Após 72h de fermentaçäo o extrato enzimático obtido pelo mutante A.niger 3T5B8 foi o que apresentou maior atividade enzimática. A atividade, medida por grupos redutores liberados e por reduçäo de viscosidade. Os valores obtidos para estes parâmetros foram, respectivamente, 56 por cento e 282 por cento maiores que os registrados para A.niger selvagem e 31 por cento e 147 por cento maiores que obtidos com o mutante A.niger 5T25A. O A.niger 3T5B8 mostrou uma acentuada reduçäo na sua capacidade de esporulaçäo e cresceu em meios com baixas atividades de água (aw) (meio básico, aw=0,96 e meio de fermentaçäo, aw=0,92), favorecendo o processo de fermentaçäo em estado sólido e formaçäo de colônias ausentes de setores em meios de cultura completo e básico

Partial purification of a polygalacturonase produced by solid-state cultures of Aspergillus niger 3T5B

A poligalacturonase produzida em cultura semi sólida de Aspergillus niger 3T5B8 foi purificada usando fracionamento por adiçäo de sal (salting out), diálise e cromatografia de gel-filtraçäo. A adiçäo de sulfato de amônio em diferentes níveis de saturaçäo apresentaram um compromisso entre purificaçäo e recuperaçäo da atividade enzimática. A cromatografia por gel-filtraç o forneceu bons resultados quando azida de sódio foi utilizada nos tampöes de eluiçäo, diminuindo as perdas na atividade enzimática em 14 por cento. O peso molecular determinado para esta poligalacturonase foi de 34700 daltons