Exopolysaccharyde production by Cryptococcus laurentii SD7 using molasses and corn steep liquor as substrates

Microbial polysaccharides are of great biotechnological and commercial interest and have wide application in the food, cosmetic and medicine industries. Exopolysaccharide (EPS) production by the yeast Cryptococcus laurentii SD7, isolated from fresh water molluscs, was studied using agro-industrial byproducts as substrates in the submerged fermentation. The Central Composite Design (CCD) 23 was used to study the influence of pH, different concentrations on sugarcane molasses and corn steep liquor (CSL), for 48 hours. According to the results, the highest EPS production occurred at the initial pH 5 and at 8.4% concentration of sugarcane molasses, which were statistically significant variable at 10% (p < 0.1). The concentration of CSL had no influence in the studied range, thus, it can be used lowest concentration (0.3%). The time course of EPS production showed that while cell growth peaked within 48 hours, the highest EPS production (6.61 g L-1) occurred at 168 hours, with a productivity of about 0.04 g L-1 h-1. The pH of the medium remained approximately constant throughout the fermentation process. The yeast C. laurentii SD7 showed great potential for EPS production at a low cost and with sustainable substrates.(AU)

Exopolysaccharyde production by Cryptococcus laurentii SD7 using molasses and corn steep liquor as substrates / Exopolysaccharyde production by Cryptococcus laurentii SD7 using molasses and corn steep liquor as substrates

Microbial polysaccharides are of great biotechnological and commercial interest and have wide application in the food, cosmetic and medicine industries. Exopolysaccharide (EPS) production by the yeast Cryptococcus laurentii SD7, isolated from fresh water molluscs, was studied using agro-industrial byproducts as substrates in the submerged fermentation. The Central Composite Design (CCD) 23 was used to study the influence of pH, different concentrations on sugarcane molasses and corn steep liquor (CSL), for 48 hours. According to the results, the highest EPS production occurred at the initial pH 5 and at 8.4% concentration of sugarcane molasses, which were statistically significant variable at 10% (p < 0.1). The concentration of CSL had no influence in the studied range, thus, it can be used lowest concentration (0.3%). The time course of EPS production showed that while cell growth peaked within 48 hours, the highest EPS production (6.61 g L-1) occurred at 168 hours, with a productivity of about 0.04 g L-1 h-1. The pH of the medium remained approximately constant throughout the fermentation process. The yeast C. laurentii SD7 showed great potential for EPS production at a low cost and with sustainable substrates.

Microbial polysaccharides are of great biotechnological and commercial interest and have wide application in the food, cosmetic and medicine industries. Exopolysaccharide (EPS) production by the yeast Cryptococcus laurentii SD7, isolated from fresh water molluscs, was studied using agro-industrial byproducts as substrates in the submerged fermentation. The Central Composite Design (CCD) 23 was used to study the influence of pH, different concentrations on sugarcane molasses and corn steep liquor (CSL), for 48 hours. According to the results, the highest EPS production occurred at the initial pH 5 and at 8.4% concentration of sugarcane molasses, which were statistically significant variable at 10% (p < 0.1). The concentration of CSL had no influence in the studied range, thus, it can be used lowest concentration (0.3%). The time course of EPS production showed that while cell growth peaked within 48 hours, the highest EPS production (6.61 g L-1) occurred at 168 hours, with a productivity of about 0.04 g L-1 h-1. The pH of the medium remained approximately constant throughout the fermentation process. The yeast C. laurentii SD7 showed great potential for EPS production at a low cost and with sustainable substrates.

Butter whey and corn steep liquor as sole raw materials to obtain a bioemulsifier from Yarrowia lipolytica for food oil-in-water emulsions / Soro de manteiga e milhocina como únicas matérias primas para obter bioemulsificante de Yarrowia lipolytica para emulsões óleo-em-água alimentícias

A synthetic medium containing glucose, glycerol, yeast extract (YE), and ammonium sulfate (AS) was compared to several low-cost media in their ability to produce high emulsification index (EI). The goal was to reduce the production costs of an emulsifier with application in food oil-in-water emulsions. To this end, agro-industrial by-products were screened for bioemulsifier production from Yarrowia lipolytica. The statistical analysis showed that the EIs of media containing residual frying oil from palm oil (RFO_palm) or soybean oil (RFO_soy), residual liquid from butter production (butter whey, BWhey) or cheese production (cheese whey, CWhey), supplemented with YE and AS were similar to the EI of the synthetic medium. The replacement of YE by corn steep liquor (CSL) also resulted in similar EI, except for RFO_soy. BWhey was tested with CSL without AS and similar EI (66.8%) was detected in comparison to that of the same medium with AS (66.3%). The cell-free broth obtained after Y. lipolytica growth in BWhey+CSL was successfully used to obtain vegetable oil-in-water emulsions indicating its potential application in food products.(AU)

O meio sintético contendo glicose, glicerol, extrato de levedura (YE) e sulfato de amônio (AS) foi comparado a meios de baixo custo para produzir alto índice de emulsificação (EI). Para este fim, os subprodutos agroindustriais foram rastreados quanto à produção de bioemulsificante por Yarrowia lipolytica. A análise estatística mostrou que EI de meios contendo óleo de fritura residual de óleo de palma (RFO_palm) ou óleo de soja (RFO_soy), líquido residual da produção de manteiga (soro de manteiga, BWhey) ou da produção de queijo (soro de queijo, CWhey), suplementado com YE e AS foram semelhantes ao EI do meio sintético. A substituição do YE por milhocina (CSL) também resultou em EI semelhante, exceto no RFO_soy. O BWhey foi testado com CSL sem AS e EI semelhante (66,8%) foi detectado em comparação com o mesmo meio com AS (66,3%). O meio isento de células obtido após o crescimento de Y. lipolytica no meio BWhey + CSL foi utilizado com sucesso para obter emulsões de óleo vegetal em água, indicando sua potencial aplicação em produtos alimentícios.(AU)

A new alkalophilic isolate of Bacillus as a producer of cyclodextrin glycosyltransferase using cassava flour

Cyclodextrin glycosyltransferase (CGTase) catalyzes the conversion of starch into non-reducing cyclic sugars, cyclodextrins, which have several industrial applications. This study aimed to establish optimal culture conditions for -CGTase production by Bacillus sp. SM-02, isolated from soil of cassava industries waste water lake. The optimization was performed by Central Composite Design (CCD) 2, using cassava flour and corn steep liquor as substrates. The maximum production of 1087.9 U mL1 was obtained with 25.0 g L1 of cassava flour and 3.5 g L1 of corn steep after 72 h by submerged fermentation. The enzyme showed optimum activity at pH 5.0 and temperature 55 °C, and maintained thermal stability at 55 °C for 3 h. The enzymatic activity was stimulated in the presence of Mg+2, Ca+2, EDTA, K+, Ba+2 and Na+ and inhibited in the presence of Hg+2, Cu+2, Fe+2 and Zn+2. The results showed that Bacillus sp. SM-02 have good potential for -CGTase production. (AU)

Lactobacillus plantarum BL011 cultivation in industrial isolated soybean protein acid residue

In this study, physiological aspects of Lactobacillus plantarum BL011 growing in a new, all-animal free medium in bioreactors were evaluated aiming at the production of this important lactic acid bacterium. Cultivations were performed in submerged batch bioreactors using the Plackett-Burman methodology to evaluate the influence of temperature, aeration rate and stirring speed as well as the concentrations of liquid acid protein residue of soybean, soy peptone, corn steep liquor, and raw yeast extract. The results showed that all variables, except for corn steep liquor, significantly influenced biomass production. The best condition was applied to bioreactor cultures, which produced a maximal biomass of 17.87 g L-1, whereas lactic acid, the most important lactic acid bacteria metabolite, peaked at 37.59 g L-1, corresponding to a productivity of 1.46 g L-1 h-1. This is the first report on the use of liquid acid protein residue of soybean medium for L. plantarum growth. These results support the industrial use of this system as an alternative to produce probiotics without animal-derived ingredients to obtain high biomass concentrations in batch bioreactors.(AU)

Atratividade de proteína hidrolisada de milho em mistura com bórax sobre moscas-das-frutas (Diptera: Tephritidae) em gaiolões de campo / Attractiveness of corn steep liquor plus borax to fruit fly (Diptera: Tephritidae) under field cages

Várias espécies de moscas-das-frutas (Diptera: Tephritidae) são capturadas em armadilhas contendo soluções aquosas preparadas com atrativos alimentares proteicos. Milhocina(r) é um produto hidrolisado de milho que apresenta um pH ácido. A adição de bórax pode aumentar o pH e melhorar a atratividade a tefritídeos adultos. Esta pesquisa objetivou avaliar a atratividade sob diferentes tempos de exposição de Milhocina(r) diluída a 5% (v/v) em mistura com 4, 7 e 10% de bórax, comparando-a com a proteína comercial padrão Bio Anastrepha a 3% (v/v). Vinte casais de Ceratitis capitata com 2 a 3 dias de idade e 20 casais de Anastrepha fraterculus com 1 a 3 dias foram liberados no interior dos gaiolões de 8,0 m3, no início de cada período de exposição (0 - 2, 2 - 4, 4 - 7 e 7 - 10 dias). No centro superior de cada gaiolão foi pendurada uma armadilha McPhail plástica transparente contendo 400 mL de solução atrativa. Ao final de cada período de exposição, os adultos capturados foram retirados, contados e sexados. Maior quantidade de adultos de C. capitata foi capturada no atrativo padrão Bio Anastrepha. Esse produto capturou mais fêmeas e machos de A. fraterculus do que os demais atrativos. Houve maiores capturas de machos e de fêmeas + machos de C. capitata do que de A. fraterculus nos tratamentos com Milhocina(r). Maiores capturas de fêmeas dos tefritídeos testados foram obtidas por Milhocina(r) + bórax 4% e por Bio Anastrepha até 7 dias de exposição dos atrativos. Em geral, no período de 2 - 4 dias de exposição houve um aumento da atratividade das soluções proteicas para fêmeas e machos de C. capitata e A. fraterculus.(AU)

Several species of fruit flies (Diptera: Tephritidae) are captured by traps baited with liquid protein food attractants. Milhocina(r) is a hydrolysate of corn with acidic pH. Addition of borax can increase the pH of the solution and improve the attractive performance for adult tephritids. The objective of this work was to evaluate the attractiveness under several times of exposure of (Milhocina(r)) at 5% (v/v) by adding borax at 4, 7 and 10% in comparison with the standard commercial protein at 3% (v/v) (BioAnastrepha). Twenty pairs of 2 to 3 years old of Ceratitis capitata and 20 pairs of 1 to 3 years old of Anastrepha fraterculus were released inside of field cage (8.0 m3) in the beginning of each exposure period (0 - 2, 2 - 4, 4 - 7 e 7 - 10 days). In the upper surface of each field cage, a transparent plastic McPhail trap was kept with 400 mL of attractive. After each exposure time, the adults captured were counted and sexed. Greater amounts of C. capitataadults were captured in attractive standard Bio Anastrepha. This product captured more females and males of A. fraterculus than other attractants. There were higher catches of male and female + male C. capitata than A. fraterculus in treatments with Milhocina(r). More captures of females tested tephritids catches were obtained by Milhocina(r) + 4% borax and Bio Anastrepha up to 7 days of exposure of the attractants. In general, during the period of 2 - 4 days of exposure, there was an increase in the attractiveness of protein solutions for females and males of C. capitata and A. fraterculus.(AU)

Attractiveness of corn steep liquor plus borax to fruit fly (Diptera: Tephritidae) under field cages / Atratividade de proteína hidrolisada de milho em mistura com bórax sobre moscas-das-frutas (Diptera: Tephritidae) em gaiolões de campo

Several species of fruit flies (Diptera: Tephritidae) are captured by traps baited with liquid protein food attractants. Milhocina® is a hydrolysate of corn with acidic pH. Addition of borax can increase the pH of the solution and improve the attractive performance for adult tephritids. The objective of this work was to evaluate the attractiveness under several times of exposure of (Milhocina®) at 5% (v/v) by adding borax at 4, 7 and 10% in comparison with the standard commercial protein at 3% (v/v) (BioAnastrepha). Twenty pairs of 2 to 3 years old of Ceratitis capitata and 20 pairs of 1 to 3 years old of Anastrepha fraterculus were released inside of field cage (8.0 m3 ) in the beginning of each exposure period (0 2, 2 4, 4 7 e 7 10 days). In the upper surface of each field cage, a transparent plastic McPhail trap was kept with 400 mL of attractive. After each exposure time, the adults captured were counted and sexed. Greater amounts of C. capitata adults were captured in attractive standard Bio Anastrepha. This product captured more females and males of A. fraterculus than other attractants. There were higher catches of male and female + male C. capitata than A. fraterculus in treatments with Milhocina®. More captures of females tested tephritids catches were obtained by Milhocina® + 4% borax and Bio Anastrepha up to 7 days of exposure of the attractants. In general, during the period of 2 - 4 days of exposure, there was an increase in the attractiveness of protein solutions for females and males of C. capitata and A. fraterculus.

Várias espécies de moscas-das-frutas (Diptera: Tephritidae) são capturadas em armadilhas contendo soluções aquosas preparadas com atrativos alimentares proteicos. Milhocina® é um produto hidrolisado de milho que apresenta um pH ácido. A adição de bórax pode aumentar o pH e melhorar a atratividade a tefritídeos adultos. Esta pesquisa objetivou avaliar a atratividade sob diferentes tempos de exposição de Milhocina® diluída a 5% (v/v) em mistura com 4, 7 e 10% de bórax, comparando-a com a proteína comercial padrão Bio Anastrepha a 3% (v/v). Vinte casais de Ceratitis capitata com 2 a 3 dias de idade e 20 casais de Anastrepha fraterculus com 1 a 3 dias foram liberados no interior dos gaiolões de 8,0 m3 , no início de cada período de exposição (0 2, 2 4, 4 7 e 7 10 dias). No centro superior de cada gaiolão foi pendurada uma armadilha McPhail plástica transparente contendo 400 mL de solução atrativa. Ao final de cada período de exposição, os adultos capturados foram retirados, contados e sexados. Maior quantidade de adultos de C. capitata foi capturada no atrativo padrão Bio Anastrepha. Esse produto capturou mais fêmeas e machos de A. fraterculus do que os demais atrativos. Houve maiores capturas de machos e de fêmeas + machos de C. capitata do que de A. fraterculus nos tratamentos com Milhocina®. Maiores capturas de fêmeas dos tefritídeos testados foram obtidas por Milhocina®+ bórax 4% e por Bio Anastrepha até 7 dias de exposição dos atrativos. Em geral, no período de 2 - 4 dias de exposição houve um aumento da atratividade das soluções proteicas para fêmeas e machos de C. capitata e A. fraterculus.

Attractiveness of corn steep liquor plus borax to fruit fly (Diptera: Tephritidae) under field cages / Atratividade de proteína hidrolisada de milho em mistura com bórax sobre moscas-das-frutas (Diptera: Tephritidae) em gaiolões de campo

Several species of fruit flies (Diptera: Tephritidae) are captured by traps baited with liquid protein food attractants. Milhocina® is a hydrolysate of corn with acidic pH. Addition of borax can increase the pH of the solution and improve the attractive performance for adult tephritids. The objective of this work was to evaluate the attractiveness under several times of exposure of (Milhocina®) at 5% (v/v) by adding borax at 4, 7 and 10% in comparison with the standard commercial protein at 3% (v/v) (BioAnastrepha). Twenty pairs of 2 to 3 years old of Ceratitis capitata and 20 pairs of 1 to 3 years old of Anastrepha fraterculus were released inside of field cage (8.0 m3 ) in the beginning of each exposure period (0 2, 2 4, 4 7 e 7 10 days). In the upper surface of each field cage, a transparent plastic McPhail trap was kept with 400 mL of attractive. After each exposure time, the adults captured were counted and sexed. Greater amounts of C. capitata adults were captured in attractive standard Bio Anastrepha. This product captured more females and males of A. fraterculus than other attractants. There were higher catches of male and female + male C. capitata than A. fraterculus in treatments with Milhocina®. More captures of females tested tephritids catches were obtained by Milhocina® + 4% borax and Bio Anastrepha up to 7 days of exposure of the attractants. In general, during the period of 2 - 4 days of exposure, there was an increase in the attractiveness of protein solutions for females and males of C. capitata and A. fraterculus.(AU)

Várias espécies de moscas-das-frutas (Diptera: Tephritidae) são capturadas em armadilhas contendo soluções aquosas preparadas com atrativos alimentares proteicos. Milhocina® é um produto hidrolisado de milho que apresenta um pH ácido. A adição de bórax pode aumentar o pH e melhorar a atratividade a tefritídeos adultos. Esta pesquisa objetivou avaliar a atratividade sob diferentes tempos de exposição de Milhocina® diluída a 5% (v/v) em mistura com 4, 7 e 10% de bórax, comparando-a com a proteína comercial padrão Bio Anastrepha a 3% (v/v). Vinte casais de Ceratitis capitata com 2 a 3 dias de idade e 20 casais de Anastrepha fraterculus com 1 a 3 dias foram liberados no interior dos gaiolões de 8,0 m3 , no início de cada período de exposição (0 2, 2 4, 4 7 e 7 10 dias). No centro superior de cada gaiolão foi pendurada uma armadilha McPhail plástica transparente contendo 400 mL de solução atrativa. Ao final de cada período de exposição, os adultos capturados foram retirados, contados e sexados. Maior quantidade de adultos de C. capitata foi capturada no atrativo padrão Bio Anastrepha. Esse produto capturou mais fêmeas e machos de A. fraterculus do que os demais atrativos. Houve maiores capturas de machos e de fêmeas + machos de C. capitata do que de A. fraterculus nos tratamentos com Milhocina®. Maiores capturas de fêmeas dos tefritídeos testados foram obtidas por Milhocina®+ bórax 4% e por Bio Anastrepha até 7 dias de exposição dos atrativos. Em geral, no período de 2 - 4 dias de exposição houve um aumento da atratividade das soluções proteicas para fêmeas e machos de C. capitata e A. fraterculus.(AU)

Atividade cicatrizante in vitro e caracterização proteica e físico-química de um gel à base de extrato aquoso de Chlorella vulgaris

A microalga Chlorella vulgaris apresenta em sua biomassa diferentes metabólitos, tais como, lipídios, polissacarídeos, enzimas, proteínas, que auxiliam na produção de agentes cicatrizantes. Um único tratamento não preenche os requisitos para ser aplicado a todos os tipos de lesões cutâneas, tendo em vista que um produto deve ser selecionado com base na profundidade, extensão, localização e origem da lesão. Desse modo, o objetivo desse trabalho foi realizar a caracterização do extrato de C. vulgaris e da formulação para a atividade cicatrizante. Inicialmente a microalga foi cultivada em meio padrão suplementado com 1% de milhocina. A biomassa seca foi ressuspendida em tampão na concentração de 50 mg/mL, sonicada e centrifugada para caracterização do sobrenadante, denominado de extrato celular. A caracterização foi realizada pela análise qualitativa de fitoquímicos por ensaio colorimétrico e para o doseamento de fenóis totais foi realizado espectroscopia e expresso por ácido gálico (AG). As proteínas presentes no extrato foram precipitadas com sulfato de amônio, purificadas por cromatografia de troca iônica e o perfil eletroforético analisado por gel SDS-PAGE. Foi realizada atividade hemaglutinante das frações proteicas obtidas utilizando eritrócitos bovinos, equinos e do sistema sanguíneo humano ABO (tipo A, B e O). A citotoxicidade in vitro do extrato celular foi testada em diferentes concentrações em fibroblastos primários. A caracterização da formulação foi realizada através de estudo organoléptico, avaliação do pH, espalhabilidade e viscosidade. Os excipientes da formulação foram submetidos à espectroscopia no infravermelho por transformada de Fourier (FT-IR) para análise da estrutura das moléculas, e ao analisador termogravimétrico (TGA) para análise da estabilidade térmica. No extrato foi detectado a presença de terpenos e conteúdo de fenóis totais de 1,35 ± 0,18 mg/g de AG. Na etapa de purificação foram obtidos dois picos, o primeiro eluído a 0,15 M de NaCl e o segundo pico eluído a 1,0 M. Apenas o primeiro pico demonstrou teor proteico de 250 g/mL, atividade hemaglutinante de 2 5 para tipo B e uma única banda com aproximadamente 36 kDa. O extrato apresentou citocompatibilidade, não induzindo efeito citotóxico nas células durante o período de 48h. A formulação apresentou cor esverdeada, textura suave, ligeiramente turva, odor característico do extrato da microalga e pH na faixa de 7,09 ± 0,07. Os resultados do teste de espalhabilidade e viscosidade indicam que a formulação pode facilmente fluir e se espalhar na área aplicada. A partir do FT-IR foi possível observar a presença de amina, carboidratos e terpenos. De acordo com a curva TGA, o percentual de perda de massa final do extrato celular foi de 23,56%, ocorrendo a 218,64 °C. carbopol teve perda de massa de 19,39%, ocorrida a 301,62%. A perda de massa da metilisotiazolinona foi de 94,69%; a curva mostra que a qualidade diminuiu rapidamente até 152,2 °C, principalmente por evaporação da água. A eficácia terapêutica das formulações tópicas é influenciada por fitoconstituintes ativos e características veiculares que desempenham um papel importante no controle da permeabilidade do medicamento. As biomoléculas ativas provenientes destes microrganismos fotossintéticos tornam-se uma alternativa potencial para o tratamento de doenças, apresentando alta disponibilidade e baixa toxicidade.

The microalgae Chlorella vulgaris presents in its biomass different metabolites such as lipids, polysaccharides, enzymes, proteins, which help in the production of healing agents. A single treatment does not meet the requirements to be applied to all types of skin lesions, since a product must be selected based on the depth, extent, location and origin of the lesion. Therefore, the objective of this work was to perform the characterization of C. vulgaris extract and the formulation for healing activity. Initially, the microalgae was cultivated in standard Bold's Basal medium supplemented with 1% corn steep liquor. The dried biomass was resuspended in a concentration of 50 mg/mL buffer, sonic and centrifuged for characterization of the supernatant, called cell extract. The characterization was performed by qualitative analysis of phytochemicals by colorimetric test and for the dosage of total phenols, spectroscopy was performed and expressed by gallic acid (AG). The proteins present in the extract were precipitated with ammonium sulfate, purified by ion exchange chromatography and the electrophoretic profile analyzed by SDS-PAGE gel. Hemagglutinating activity of the protein fractions obtained using bovine, equine and human ABO (type A, B and O) red blood cells was performed. The in vitro cytotoxicity of the cell extract was tested in different concentrations in primary fibroblasts. The characterization of the formulation was performed through organoleptic study, evaluation of pH, spreadability and viscosity. The excipients of the formulation were submitted to Fourier transform infrared spectroscopy (FT-IR) for analysis of the structure of the molecules, and to a thermogravimetric analyzer (TGA) for thermal stability analysis. Terpenes and total phenol content of 1.35 ± 0.18 mg/g AG were detected in the extract. In the purification step two peaks were obtained, the first eluate at 0.15 M of NaCl and the second at 1.0 M. Only the first peak showed protein content of 250 g/mL, hemagglutinating activity of 25 for type B and a single band with approximately 36 kDa. The extract showed cytocompatibility, not inducing cytotoxic effect in the cells during the 48h period. The formulation had a greenish color, smooth texture, slightly cloudy, characteristic odor of the microalgae extract and pH in the range of 7.09 ± 0.07. The results of the spreadability and viscosity test indicate that the formulation can easily flow and spread over the applied area. From the FT-IR it was possible to observe the presence of amine, carbohydrates and terpenes. According to the TGA curve, the percentage of final cell extract mass loss was 23.56%, occurring at 218.64 °C. carbopol had a mass loss of 19.39%, occurring at 301.62%. The loss in mass of methylisothiazolinone was 94.69%; the curve shows that the quality decreased rapidly to 152.2 °C, mainly by evaporation of water. The therapeutic efficacy of topical formulations is influenced by active phytoconstituents and vehicular characteristics that play an important role in the control of the drug permeability. The active biomolecules from these photosynthetic microorganisms become a potential alternative for the treatment of diseases, presenting high availability and low toxicity

Extracellular production of avicelase by the thermophilic soil bacterium Bacillus sp. SMIA-2 / Produção de avicelase extracelular pela bactéria termofílica Bacillus sp. SMIA-2

Nowadays, the isolation of new bacterial strains that produce enzymes with novel properties is a subject of great relevance to the scientific community. This study, in order to search for producers of new cellulase strains, investigated the avicelase production by thermophilic Bacillus sp. strain SMIA-2. The best avicelase activity was observed in a culture medium containing 0.5% (w v-1) avicel and 0.5% (w v-1) corn steep liquor with initial pH 7.5-8.0 incubated at 50oC. When avicel was replaced in the medium by the treated sugarcane bagasse (0.5%, w v-1) the avicelase activity levels were not affected. Studies on the avicelase characterization revealed that the optimum pH of the enzyme was found to be 8.5 and the enzyme retained more than 80% of its activity after incubation at room temperature for 2h at pH 6.5-8.5. The optimum temperature of this enzyme was 70oC and the enzyme retained 67% of the original activity after 20 min. of heat treatment at 70oC. Avicelase was stimulated by Mn2+ and Co2+, whereas Hg2+ greatly inhibited the enzyme activity.(AU)

Atualmente, o isolamento de estirpes de bactérias que produzem enzimas com novas propriedades é um tema de grande relevância para a comunidade científica. Este trabalho, buscando por novas cepas produtoras de celulases, investigou a produção de avicelases pelo termofílico Bacillus sp. cepa SMIA-2. A melhor atividade da enzima foi obtida em uma cultura contendo 0,5% (p v-1) avicel e 0,5% (p v-1) água de maceração de milho com pH inicial de pH 7,5-8,0 e incubada a 50oC. A substituição da avicel no meio de cultura pelo bagaço de cana- de- açúcar tratado (0,5%, p v-1) não afetou os níveis de atividade da avicelase. Estudos sobre a caracterização da avicelase revelaram que o pH para atividade ótima da enzima foi 8,5 e que a mesma reteve mais de 80% de sua atividade após ser incubada à temperatura ambiente por 2 h a pH 6,5- 8,5. A temperatura ótima da avicelase foi 70oC e a enzima reteve 67% da sua atividade original após 20 min. de incubação a 70oC. A avicelase foi estimulada pelos íons Mn2+ e Co2+, ao passo que Hg2+ inibiu a atividade da enzima.(AU)

Kinetic characterization of glucose aerodehydrogenase from Aspergillus niger EMS-150-F after optimizing the dose of mutagen for enhanced production of enzyme

In the present study enhanced production of glucose aerodehydrogenase from Aspergillus niger has been achieved after optimizing the dose of chemical mutagen ethyl methane sulfonate (EMS) that has not been reported earlier. Different doses of mutagen were applied and a strain was developed basing upon the best production. The selected strain Aspergillus niger EMS-150-F was optimized for nutrient requirements in order to produce enzyme through fermentation and the results showed the best yield at 2% corn steep liquor (CSL), 36 hours fermentation time, pH 5, 30°C temperature, 0.3% KH2PO4, 0.3% urea and 0.06% CaCO3. The enzyme was then purified and resulted in 57.88 fold purification with 52.12% recovery. On kinetic characterization, the enzyme showed optimum activity at pH 6 and temperature 30°C. The Michaelis-Menton constants (Km, Vmax, Kcat and Kcat/Km) were 20 mM, 45.87 U mL-1, 1118.81 s-1 and 55.94 s-1 mM-1, respectively. The enzyme was found to be thermaly stable and the enthalpy and free energy showed an increase with increase in temperature and S* was highly negative proving the enzyme from A. niger EMS-150-F resistant to temperature and showing a very little disorderliness.

Produção de biossurfactante por Chromobacterium violaceum ATCC 12472 utilizando milhocina e óleo de milho pós-fritura como nutrientes

The interest in microbial surfactants has been increased considerably in the recent years. Environmental concern among consumers, combined with new laws control of the environment, the demand for natural surfactants, to replace petrochemicals. The microbial surfactants have been tested in many industrial and environmental applications, such as in bioremediation, oil dispersion and oil recovery, replacing chemical surfactants. Furthermore, they can also be used in the food, cosmetic, detergent and agriculture. However, increased environmental concern among consumers to new legislation combined control of the environment has led to demand for natural surfactants as alternatives to existing products. This work aimed to produce biosurfactant by Chromobacterium violaceum ATCC 12472, using low nutrient value. The surface tension of the biosurfactant produced by C. violaceum ATCC 12472 had values between 29 and 40 mN/m. The best result was observed on the assay 8 (8.0% of corn steep liquor, 1.0% of lactose and corn oil post-frying 7.5%) factorial design, within 72 hours of fermentation, lowering the surface tension of water from 71 to 29 mN/m. The best results of the index (E24) and emulsification activity (UEA) were observed in the central point, and using the both methods and corn post-frying oil resulted in 72% of emulsification, and the emulsification activity value of 6 UAE. The results demonstrate the high potential of C. violaceum promising in producing a biosurfactant.

O interesse em surfactantes de origem microbiana tem aumentado consideravelmente nos últimos anos. A preocupação ambiental entre os consumidores, combinada às novas legislações de controle do meio ambiente, aumentou a procura por surfactantes naturais em substituição aos derivados petroquímicos. Os surfactantes microbianos têm sido testados em muitas aplicações ambientais e industriais, como na biorremediação, na dispersão de manchas oleosas e na recuperação de petróleo, substituindo os surfactantes químicos. Além disso, também podem ser utilizados nas indústrias de alimentos, cosméticos, detergentes e na agricultura. Neste trabalho foi produzido biossurfactante por Chromobacterium violaceum ATCC 12472, utilizando nutrientes de baixo valor agregado. A tensão superficial do biossurfactante produzido por C. violaceum ATCC 12472 teve valores entre 29 e 40 mN/m. O melhor resultado foi observado no ensaio 8 (milhocina 8,0%, lactose 1,0% e óleo de milho pós-fritura 7,5%) do planejamento fatorial, com 72 horas de fermentação, reduzindo a tensão superficial da água de 71 para 29 mN/m. Os melhores resultados do índice (E24) e da atividade de emulsificação (UEA) foram observados no ponto central, sendo utilizados para ambos os métodos o óleo de milho, que resultou no E24 de 2% e atividade com valores acima de 6 UAE. Os resultados obtidos demonstram o elevado potencial de C. violaceum na produção de um promissor biossurfactante.

Biosurfactant production by Chr omobacterium violaceum ATCC 12472 using corn steep liquor and corn post-frying oil as nutrients / Produção de biossurfactante por Chromobacterium violaceum ATCC 12472 utilizando milhocina e óleo de milho pós-fritura como nutrientes

The interest in microbial surfactants has been increased considerably in the recent years. Environmental concern among consumers, combined with new laws control of the environment, the demand for natural surfactants, to replace petrochemicals. The microbial surfactants have been tested in many industrial and environmental applications, such as in bioremediation, oil dispersion and oil recovery, replacing chemical surfactants. Furthermore, they can also be used in the food, cosmetic, detergent and agriculture. However, increased environmental concern among consumers to new legislation combined control of the environment has led to demand for natural surfactants as alternatives to existing products. This work aimed to produce biosurfactant by Chromobacterium violaceum ATCC 12472, using low nutrient value. The surface tension of the biosurfactant produced by C. violaceum ATCC 12472 had values between 29 and 40 mN/m. The best result was observed on the assay 8 (8.0% of corn steep liquor, 1.0% of lactose and corn oil post-frying 7.5%) factorial design, within 72 hours of fermentation, lowering the surface tension of water from 71 to 29 mN/m. The best results of the index (E24) and emulsification activity (UEA) were observed in the central point, and using the both methods and corn post-frying oil resulted in 72% of emulsification, and the emulsification activity value of 6 UAE. The results demonstrate the high potential of C. violaceum promising in producing a biosurfactant.

O interesse em surfactantes de origem microbiana tem aumentado consideravelmente nos últimos anos. A preocupação ambiental entre os consumidores, combinada às novas legislações de controle do meio ambiente, aumentou a procura por surfactantes naturais em substituição aos derivados petroquímicos. Os surfactantes microbianos têm sido testados em muitas aplicações ambientais e industriais, como na biorremediação, na dispersão de manchas oleosas e na recuperação de petróleo, substituindo os surfactantes químicos. Além disso, também podem ser utilizados nas indústrias de alimentos, cosméticos, detergentes e na agricultura. Neste trabalho foi produzido biossurfactante por Chromobacterium violaceum ATCC 12472, utilizando nutrientes de baixo valor agregado. A tensão superficial do biossurfactante produzido por C. violaceum ATCC 12472 teve valores entre 29 e 40 mN/m. O melhor resultado foi observado no ensaio 8 (milhocina 8,0%, lactose 1,0% e óleo de milho pós-fritura 7,5%) do planejamento fatorial, com 72 horas de fermentação, reduzindo a tensão superficial da água de 71 para 29 mN/m. Os melhores resultados do índice (E24) e da atividade de emulsificação (UEA) foram observados no ponto central, sendo utilizados para ambos os métodos o óleo de milho, que resultou no E24 de 2% e atividade com valores acima de 6 UAE. Os resultados obtidos demonstram o elevado potencial de C. violaceum na produção de um promissor biossurfactante.

Biosurfactant production by Chr omobacterium violaceum ATCC 12472 using corn steep liquor and corn post-frying oil as nutrients / Produção de biossurfactante por Chromobacterium violaceum ATCC 12472 utilizando milhocina e óleo de milho pós-fritura como nutrientes

The interest in microbial surfactants has been increased considerably in the recent years. Environmental concern among consumers, combined with new laws control of the environment, the demand for natural surfactants, to replace petrochemicals. The microbial surfactants have been tested in many industrial and environmental applications, such as in bioremediation, oil dispersion and oil recovery, replacing chemical surfactants. Furthermore, they can also be used in the food, cosmetic, detergent and agriculture. However, increased environmental concern among consumers to new legislation combined control of the environment has led to demand for natural surfactants as alternatives to existing products. This work aimed to produce biosurfactant by Chromobacterium violaceum ATCC 12472, using low nutrient value. The surface tension of the biosurfactant produced by C. violaceum ATCC 12472 had values between 29 and 40 mN/m. The best result was observed on the assay 8 (8.0% of corn steep liquor, 1.0% of lactose and corn oil post-frying 7.5%) factorial design, within 72 hours of fermentation, lowering the surface tension of water from 71 to 29 mN/m. The best results of the index (E24) and emulsification activity (UEA) were observed in the central point, and using the both methods and corn post-frying oil resulted in 72% of emulsification, and the emulsification activity value of 6 UAE. The results demonstrate the high potential of C. violaceum promising in producing a biosurfactant. (AU)

O interesse em surfactantes de origem microbiana tem aumentado consideravelmente nos últimos anos. A preocupação ambiental entre os consumidores, combinada às novas legislações de controle do meio ambiente, aumentou a procura por surfactantes naturais em substituição aos derivados petroquímicos. Os surfactantes microbianos têm sido testados em muitas aplicações ambientais e industriais, como na biorremediação, na dispersão de manchas oleosas e na recuperação de petróleo, substituindo os surfactantes químicos. Além disso, também podem ser utilizados nas indústrias de alimentos, cosméticos, detergentes e na agricultura. Neste trabalho foi produzido biossurfactante por Chromobacterium violaceum ATCC 12472, utilizando nutrientes de baixo valor agregado. A tensão superficial do biossurfactante produzido por C. violaceum ATCC 12472 teve valores entre 29 e 40 mN/m. O melhor resultado foi observado no ensaio 8 (milhocina 8,0%, lactose 1,0% e óleo de milho pós-fritura 7,5%) do planejamento fatorial, com 72 horas de fermentação, reduzindo a tensão superficial da água de 71 para 29 mN/m. Os melhores resultados do índice (E24) e da atividade de emulsificação (UEA) foram observados no ponto central, sendo utilizados para ambos os métodos o óleo de milho, que resultou no E24 de 2% e atividade com valores acima de 6 UAE. Os resultados obtidos demonstram o elevado potencial de C. violaceum na produção de um promissor biossurfactante. (AU)

Penicillin production by wild isolates of Penicillium chrysogenum in Pakistan

The present study was aimed at exploring the native wild isolates of Penicillium chrysogenum series in terms of their penicillin production potential. Apart from the standard medium, the efforts were made to utilize suitable agro-industrial wastes for the maximum yield of penicillin. Two series of P. chrysogenum were isolated from local sources and named as P. chrysogenum series UAF R1 and P. chrysogenum series UAF R2. The native series were found to possess better penicillin production potential than the already reported series of P. chrysogenum. However, P. chrysogenum series UAF R1 was found to be the best candidate for high yield of penicillin starting at 100 hour as compared to P. chrysogenum series UAF R2 which produced the highest yield of penicillin at 150 hours for a shorter period of time. Addition of Corn Steep Liquor (CSL) to the fermentation medium resulted in the production of 1.20g/L penicillin by P. chrysogenum series UAF R1 and P. chrysogenum series UAF R2. The fermentation medium in which Sugar Cane Bagasse (SCB) was replaced with CSL resulted in the highest yield of penicillin (1.92g/L) by both native series of P. chrysogenum. The penicillin production was increased by 62.5% in medium with SCB as compared to that with CSL. The penicillin yield of medium containing lactose and phenyl acetate was higher than that of control medium. Overall results revealed that P. chrysogenum series UAF R1 and P. chrysogenum series UAF R2 may be recommended for better yield of natural penicillin and this efficiency may be further enhanced by utilizing SCB as substrate in the growth medium.

Brewer's spent grain and corn steep liquor as alternative culture medium substrates for proteinase production by Streptomyces malaysiensis AMT-3

Brewer's spent grain and corn steep liquor or yeast extract were used as the sole organic forms for proteinase production by Streptomyces malaysiensis in submerged fermentation. The influence of the C and N concentrations, as well as the incubation periods, were assessed. Eight proteolytic bands were detected through gelatin-gel-electrophoresis in the various extracts obtained from the different media and after different incubation periods, with apparent molecular masses of 20, 35, 43, 50, 70, 100, 116 and 212 kDa. The results obtained suggest an opportunity for exploring this alternative strategy for proteinases production by actinomycetes, using BSG and CSL as economically feasible substrates.

Kinetics of high-Level of ß-glucosidase production by a 2-deoxyglucose-resistant mutant of Humicola lanuginosa in submerged fermentation

A 2-deoxyglucose-resistant mutant (M7) of Humicola lanuginosa was obtained by exposing conidia to -rays and permitting expression in broth containing 0.6% 2-deoxyglucose (DG) and cellobiose (1%) before plating on DG esculin-ferric ammonium citrate agar medium from which colonies showing faster and bigger blackening zones were selected. Kinetic parameters for enhanced ß-glucosidase (BGL) synthesis by M7 were achieved when corncobs acted as the carbon source. The combination between corncobs and corn steep liquor was the best to support higher values of all product formation kinetic parameters. Effect of temperature on the kinetic and thermodynamic attributes of BGL production equilibrium in the wild organismand M7was studied using batch process at eight different temperatures in shake-flask studies. The best performance was found at 45ºC and 20 g L-1 corncobs in 64 h. Both growth and product formation (17.93 U mL-1) were remarkably high at 45ºC and both were coupled under optimum working conditions. Product yield of BGL from the mutant M7 (1556.5 U g-1 dry corncobs) was significantly higher than the values reported on all fungal and bacterial systems. Mutation had thermo-stabilization influence on the organism and mutant required lower activation energy for growth and lower magnitudes of enthalpy and entropy for product formation than those demanded by the wild organism, other mesophilic and thermo-tolerant organisms. In the inactivation phase, the organisms needed lower values of activation energy, enthalpy and entropy for product formation equilibrium, confirming thermophilic nature of metabolic network possessed by the mutant organism.

Um mutante de Hemicola lanuginosa resistente a 2-deoxiglucose(M7) foi obtido através de exposição de conídios a raios , permitindo a expressão em caldo contendo 0,6% de 2-deoxiglucose (DG) e celobiose (1%) antes da semeadura em ágar DG esculina citrato de ferro amoniacal, da qual foram selecionadas as colônias com halo negro. Os parâmetros cinéticos para produção aumentada de ß-glucosidase (BGL) foram obtidos empregando-se sabugo de milho como fonte de carbono. A combinação de espiga de milho com água de maceração de milho foi a que forneceu os valores mais altos nos parâmetros cinéticos de formação de todos os produtos. O efeito da temperatura na cinética e atributos termodinâmicos da produção de BGL pelas cepas selvagem e M7 foi avaliado empregando-se processo de batelada em oito temperaturas diferentes in frascos em agitação. O melhor desempenho foi observado a 45ºC e 20g.l-1 de espiga de milho em 64h. Tanto a multiplicação quanto a formação do produto foram muito altas a 45ºC e ambas estavam ligadas em condições ótimas de trabalho. O rendimento de BGL produzido pelo mutante M7 (1556 U.g-1 de espiga seca) foi significativamente superior aos valores reportados para todos os sistemas fúngicos e bacterianos. A mutação influenciou a termoestabilização no microrganismo, sendo que o mutante necessitou de energia de ativação mais baixa para multiplicação e valores mais baixos de entalpia e entropia para a formação do produto quando comparado à cepa selvagem e a outros microrganismos mesofilicos e termotolerantes. Na fase de inativação, os microrganismos necessitaram valores mais baixos de energia de ativação, entalpia e entropia para o equilíbrio da formação de produto, confirmando a natureza termofílica da máquina metabólica do mutante.

Effect of the culture conditions on the production of an extracellular protease by thermophilic Bacillus sp and some properties of the enzymatic activity

Protease production by thermophilic Bacillus sp strain SMIA-2 cultivated in liquid cultures containing 1% maltose as a carbon source and supplemented with whey protein (0.1%) and corn steep liquor (0.3%) reached a maximum at 14 h, with levels of 42 U/mg protein. The microorganism was capable of utilizing a wide range of carbon sources, but protease activity varied according the carbon source. Starch and maltose were the best carbon sources in the present study for protease secretion, while lactose and sucrose were less effective. Increasing maltose concentration in the medium until 1%, improved the growth of the organism and the enzyme activity. Regarding the amounts of corn steep liquor and whey protein in the medium, the concentrations of 0.2% and 0.1% respectively, were considered the most effective for protease secretion by the organism. Studies on the protease characterization revealed that the optimum temperature of this enzyme was 70ºC. Thermostability profile indicated that the enzyme retained 80% of the original activity after 2 h heat treatment at 60ºC. At 70ºC, 70% of the original activity was retained after 15 min heat treatment. The optimum pH of the enzyme was found to be 8.5. After incubation of crude enzyme solution at room temperature for 2 h at pH 6.0-10.0, a decreased of about 15% of its original activity at pH 8.5 was observed. At pH 10.0, the decrease was 24%. In the presence of 1.0 M and 5.0 M NaCl, 76% and 37% of protease activity was retained after 2 h incubating at 45ºC respectively.

A produção de proteases pelo termofílico Bacillus sp cepa SMIA-2 cultivado em culturas líquidas contendo maltose (1%) e suplementada com proteínas de soro (0,1%) e água de maceração de milho (0,3%) alcançou o máximo em 14 h, com níveis de 42 U/mg proteína. O microrganismo foi capaz de utilizar várias fontes de carbono, mas a atividade da protease variou com cada fonte. Amido e maltose foram as melhores fontes para a secreção da protease, enquanto lactose e sacarose não foram muito efetivas. O aumento da concentração de maltose no meio de cultura até 1% melhorou o crescimento do organismo e a atividade da enzima. Em relação à concentração de proteínas do soro e da água de maceração de milho no meio de cultura, 0,1% e 0,2% respectivamente, foram as mais efetivas para a secreção da enzima pelo organismo. Estudos sobre a caracterização da protease revelaram que a temperatura ótima desta enzima foi 70ºC. Em relação a termoestabilidade da enzima, a protease manteve 80% de sua atividade original após 2 h de tratamento a 60ºC. A 70ºC, 70% de sua atividade original foi mantida após 15 min. O pH ótimo para atividade da enzima foi 8,5. Após a incubação da solução enzimática bruta a pH 6,0-10.0 por 2 h a temperatura ambiente, foi observado um decréscimo de em torno de 15% da sua atividade original a pH 8,5. A pH 10,0 o decréscimo na atividade foi de 24%. Na presença de 1.0 M e 5.0 M NaCl, 76% e 37% da atividade da protease foi mantida após 2 h de incubação a 45ºC respectivamente.

Media components and amino acid supplements influencing the production of fruity aroma by Geotrichum candidum

The ability of Geotrichum candidum to produce fruity aroma in food grade sucrose, molasses, corn steep liquor and peptone based culture media was tested by sensory evaluation and analyzed by gas chromatography mass spectrometry. A strong and sweet fruity aroma was produced from molasses, with peptone or corn steep liquor stimulating aroma production. Molasses with peptone supplemented with leucine, valine, or alanine yielded better fruity aroma production and the presence of many esters was consistent with the fruity aroma production.

Geotrichum candidum foi cultivado em diversos meios de cultura contendo sacarose ou melaço e milhocina ou peptona e a produção de aroma frutal foi verificada através de avaliação sensorial e cromatografia gasosa acoplada a espectrometria de massas. Os meios contendo melaço, peptona e leucina, valina ou alanina apresentaram os melhores resultados e a presença de diversos ésteres foi consistente com a formação de aroma frutal.

Production of glucosyltransferase by Erwinia sp. using experimental design and response surface methodology

Glucosyltransferase produced by strain Erwinia sp. is an intracellular enzyme that catalyzes the formation of isomaltulose from sucrose. Isomaltulose is a non-cariogenic reducing dissacharide commercially used in foods. Response surface methodology and 2³-factorial central composite design were employed to optimize a fermentation medium for the production of glucosyltransferase by Erwinia sp. in shaken flasks at 200 rpm and 30ºC. The three variables involved in this study were sugar cane molasses (SCM), corn steep liquor (CSL) and yeast extract Prodex Lac SD (YEP). The statistical analysis of the results showed that, in the range studied, all the factors had a significant effect on glucosyltransferase production and the optimum medium composition for enzyme production was (in g l-1) SCM-100, CSL-60 and YEP-8, which lead to a glucosyltransferase activity of 6.65 U mL-1.

A glicosiltransferase obtida pela linhagem Erwinia sp. é uma enzima intracelular que catalisa a conversão de sacarose em isomaltulose. A isomaltulose é um dissacarídeo redutor, não cariogênico e comercialmente utilizado em alimentos como substituto da sacarose. A metodologia de superfície de resposta e planejamento fatorial composto central-2³ foram utilizados para otimizar o meio de cultivo para a produção de glicosiltransferase de Erwinia sp. em frascos sob agitação a 200 rpm e 30ºC. As três variáveis independentes envolvidas no estudo foram o melaço de cana de açúcar, a água de maceração de milho e o extrato de levedura Prodex Lac SD. As análises estatísticas dos resultados mostraram que, dentro da faixa estudada das concentrações dos componentes de meio de cultivo, todas as variáveis apresentaram efeito significativo na produção de glicosiltransferase. O meio de cultivo otimizado foi composto de 100 gL-1 de melaço de cana de açúcar, 60 gL-1 de água de maceração de milho e 8 gL-1 de extrato de levedura Prodex Lac SD, apresentando atividade de glicosiltransferase de 6.65 U mL-1.