Different culture conditions viz. additional carbon and nitrogen content, inoculum size and age, temperature and pH of the mixed culture of Bifidobacterium bifidum and Lactobacillus acidophilus were optimized using response surface methodology (RSM) and artificial neural network (ANN). Kinetic growth models were fitted for the cultivations using a Fractional Factorial (FF) design experiments for different variables. This novel concept of combining the optimization and modeling presented different optimal conditions for the mixture of B. bifidum and L. acidophilus growth from their one variable at-a-time (OVAT) optimization study. Through these statistical tools, the product yield (cell mass) of the mixture of B. bifidum and L. acidophilus was increased. Regression coefficients (R2) of both the statistical tools predicted that ANN was better than RSM and the regression equation was solved with the help of genetic algorithms (GA). The normalized percentage mean squared error obtained from the ANN and RSM models were 0.08 and 0.3%, respectively. The optimum conditions for the maximum biomass yield were at temperature 38°C, pH 6.5, inoculum volume 1.60 mL, inoculum age 30 h, carbon content 42.31% (w/v), and nitrogen content 14.20% (w/v). The results demonstrated a higher prediction accuracy of ANN compared to RSM.

The culture conditions viz. additional carbon and nitrogen content, inoculum size, age, temperature and pH of Lactobacillus acidophilus were optimized using response surface methodology (RSM) and artificial neural network (ANN). Kinetic growth models were fitted to cultivations from a Box-Behnken Design (BBD) design experiments for different variables. This concept of combining the optimization and modeling presented different optimal conditions for L. acidophilus growth from their original optimization study. Through these statistical tools, the product yield (cell mass) of L. acidophilus was increased. Regression coefficients (R²) of both the statistical tools predicted that ANN was better than RSM and the regression equation was solved with the help of genetic algorithms (GA). The normalized percentage mean squared error obtained from the ANN and RSM models were 0.06 and 0.2%, respectively. The results demonstrated a higher prediction accuracy of ANN compared to RSM.

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.

The aim of this work was to apply a modeling integrated optimisation approach for a complex, highly nonlinear system for an extracellular lipase extraction process. The model was developed using mutation, crossover and selection variables of Differential Evolution (DE) based on central composite design of Response Surface Methodology. The experimentally validated model was optimized by DE, a robust evolutionary optimization tool. A maximum lipase activity of 134.13 U/gds (more than 36.28 U/gds compared to one variable at a time approach) was observed with the DE-stated optimum values of 25.01% dimethyl sulfoxide concentration, 40 mM buffer, 128.52 min soaking time and 35ºC with the DE control parameters, namely number of population, generations, crossover operator and scaling factor as 20, 50, 0.5 and 0.25, respectively. The use of DE approach improved the optimization capability and decision speed, resulting in an improved yield of 36.28 U/gds compared to the one variable at a time approach for the extracellular lipase activity under the non-optimized conditions. The developed mathematical model and optimization were generic in nature, which seemed to be useful for the scale-up studies of maximum recovery of lipase from the fermented biomass.

The aim of this work was to optimize the biomass production by Bifidobacterium bifidum 255 using the response surface methodology (RSM) and artificial neural network (ANN) both coupled with GA. To develop the empirical model for the yield of probiotic bacteria, additional carbon and nitrogen content, inoculum size, age, temperature and pH were selected as the parameters. Models were developed using ¼ fractional factorial design (FFD) of the experiments with the selected parameters. The normalized percentage mean squared error obtained from the ANN and RSM models were 0.05 and 0.1%, respectively. Regression coefficient (R²) of the ANN model showed higher prediction accuracy compared to that of the RSM model. The empirical yield model (for both ANN and RSM) obtained were utilized as the objective functions to be maximized with the help of genetic algorithm. The optimal conditions for the maximal biomass yield were 37.4 °C, pH 7.09, inoculum volume 1.97 ml, inoculum age 58.58 h, carbon content 41.74% (w/v), and nitrogen content 46.23% (w/v). The work reported is a novel concept of combining the statistical modeling and evolutionary optimization for an improved yield of cell mass of B. bifidum 255.

Optimization of lipase production by Enterobacter aerogenes was carried out using response surface methodology (RSM) where the statistical model was obtained by fractional factorial central composite design. The influence of various physico-chemical parameters, viz. temperature, oil concentration, inoculum volume, pH and incubation period on lipase production was examined. Optimization of physico-chemical parameters resulted 1.4- fold increase in lipase activity. The optimum levels of parameters were 34°C, oil concentration 3%, inoculum volume 7%, pH 7 and incubation time 60 h for obtaining a maximum lipase activity of 27.25 U/ml.

The aim of this study was to evaluate the interaction effects of the physico-chemical parameters on the endoglucanase (CMCase) production by Trichoderma reesei Rut C30 on a cellulosic agro-residue by the solid-state fermentation (SSF) and to determine their optimum values by the EVOP factorial design technique. The best combination of physical parameters for the maximum production of the endoglucanase (CMCase) was 28ºC temperature, 79% relative humidity and 4.8 pH of the medium. The best combination of the chemical parameters was (mg/L) nicotinic acid 15, naphthalene acetic acid 7, ferric chloride 5 and Tween-80 6. With the application of this technique, the yield of the CMCase increased by ~ 2.3 fold.

Uma bactéria isolada do solo e identificada como Pseudomonas sp. RAJR 044 demonstrou ser uma potencial produtora de protéase extracelular. Um mutante JNGR 242 dessa espécie foi obtido mediante radiação ultravioleta sob condições experimentalmente otimizadas de pH 7,0, temperatura de 34ºC, volume de inóculo de 1,0 mL e tempo de incubação de 24 hora produziu 2,5 mais protease. Também foram realizadas análises comparativas das características química quanto assimilação de diferentes fontes de carbono e de nitrogênio. O crescimento máximo desse mutante na placa de ágar gelatina (2%) foi obtido na presença de sacarose (2%), maltose (2%), sulfato de amônia (2%) e nitrato de potássio (2%), ao passo que no caso da linhagem original a melhor fonte de carbono foi a sacarose (2%) e o nitrato de amônia (2%) como a melhor fonte de nitrogênio. As protéases purificadas de ambas as espécies (original e mutante) mostraram faixas homogêneas que correspondem ao peso molecular 14,4 kDa na SDS-PAGE. Ao se estudar as propriedades cinéticas de ambas as espécies, observou-se que a taxa de hidrólise da caseína era máxima em pH 7,0 e 8,0, , temperaturas de 45ºC e 60ºC para a espécie original e a mutante, respectivamente. Observou-se também que ambas as protéases extracelulares foram inibidas pela serina, isto é, PMSF à concentração de 2mM.

A potent bacterium for extracellular protease production was isolated from local soil and identified as Pseudomonas sp. RAJR 044. A mutant of this strain JNGR 242 with protease productivity 2.5 fold higher was obtained by ultraviolet irradiation under experimentally optimized conditions of pH 7.0, temperature of 34ºC, inoculum volume of 1.0 mL and incubation time of 24 hours. Comparative analysis of the chemical characteristics i.e. assimilation of carbon and nitrogen sources were also carried out. Maximum growth of the mutant strain in 2% gelatin agar plate was obtained in presence of dextrose (2%), maltose (2%), ammonium sulfate (2%) and potassium nitrate (2%) whereas, that of the parent strain was found in sucrose (2%) and ammonium nitrate (2%). The purified proteases from both the strains (parent and mutant) appeared as single homogeneous bands corresponding to 14.4 kDa molecular weight on SDS-PAGE. On studying the kinetic properties of both strains it was observed that the rate of casein hydrolysis was maximum at pH 8.0 and 7.0 and temperatures 45º C and 60º C for the parent and mutant strains respectively. It was also observed that both the extracellular proteases were inhibited by a serine protease inhibitor i.e. PMSF at 2mM concentration.

Um maltooligossacarideo obtido a partir de amilase produzida por B. circulans GRS 313 foi imobilizada em alginato de sódio. A atividade enzimática foi afetada pelo tamanho da partícula. Partículas com 2mm foram as mais efetivas na hidrólise. Constantes cinética Km e Vmax foram estimadas e afetadas pelo tamanho das partículas. A atividade catalítica da enzima foi estuda na presença de diferentes tipos de amido e íons metálicos. HgCl2, CuSO4 e FeCl3 provocaram inibição na enzima. As condições de reação (temperatura e pH) foram otimizadas utilizando a metodologia da superfície de resposta. Em pH ótimo de 4,9 e temperatura de 57 ºC, a atividade aparente foi de 25.6 U/g de partículas, resultando num acréscimo de mais de 2 vezes na atividade da enzima. A imobilização da enzima mostrou uma alta estabilidade operacional pela retenção de 85% de sua atividade inicial após sete ciclos de utilização.

A maltooligosaccharide-forming amylase from B circulans GRS 313 was immobilized by entrapment in calcium alginate beads. The immobilized activity was affected by the size of the bead and bead size of 2mm was found to be most effective for hydrolysis. Kinetics constants, Km and Vmax were estimated and were found to be affected by the bead size. The catalytic activity of the enzyme was studied in presence of various starchy residues and metal ions. HgCl2, CuSO4 and FeCl3 caused inhibition of the enzyme. The reaction conditions, pH and temperature, was optimized using response surface methodology. At the optimum pH and temperature of 4.9 and 57ºC, the apparent activity was 25.6U/g of beads, resulting in almost 2-fold increase in activity. The immobilized enzyme showed a high operational stability by retaining almost 85% of the initial activity after seventh use.

Otimizou-se a extração de alfa- amilase na fermentação no estado sólido do farelo de trigo Bacillus circulans GRS313 . Entre vários solventes testados, conseguiu-se a melhor extração com solução 2,5% (v/v) de glicerol. As condições ótimas de extração foram de 2,5 horas de tempo de contato a 30° C sob condições de agitação e recirculação, chegando a uma atividade enzimática de 590 U/g farelo, enquanto chegou-se a uma atividade enzimática de 439.58U/g farelo sob condições de contato estacionárias. Com lavagens repetidas nas condições ótimas descritas acima, chegou-se à uma atividade enzimática de 600 U/g farelo e 184,2 U/g de enzima extraída nas 1 ª e 2ª lavagens, respectivamente, e apenas 36,84 U/g recuperada na 3 ª lavagem.

The extraction of alpha-amylase in the solid state fermentation of wheat bran by Bacillus circulans GRS313 was optimized. Among various solvents tested, maximum extraction was achieved when 2.5% (v/v) glycerol was added. The optimum conditions for extraction were 2.5 hrs soaking time at 30° C under recirculation and agitated condition, which yielded 590 U/g bran of enzyme activity. Whereas under stationary soaking condition the activity of the enzyme was 439.58U/g bran. With repeated wash under the above optimum conditions showed that 600 U/g and 184.2 U/g of enzyme extracted in the 1st and 2nd washes respectively and only 36.84 U/g was recovered in the 3rd wash

Para predizer a taxa das reações enzimaticas uma correlação empírica baseada nos resultados experimentais foi desenvolvida. Os modelos representam a ativação e a desativativação da hydrolise enzimatica. Os resultados foram avaliados pela análise de variança (ANOVA). A atividade máxima da tannase foi obtida após 5 minutos de incubação, temperatura 40ºC, pH 4,0, concentração inicial da enzima de 0,12 v/v, concentração inicial do substrato 0,42 mg/ml, força iônica 0,2 M. Sob essas condições a taxa máxima de produção ácido galico foi de 33,49 µmoles/ml/min.

For predicting the rate of enzymatic reaction empirical correlation based on the experimental results obtained under various operating conditions have been developed. Models represent both the activation as well as deactivation conditions of enzymatic hydrolysis and the results have been analyzed by analysis of variance (ANOVA). The tannase activity was found maximum at incubation time 5 min, reaction temperature 40ºC, pH 4.0, initial enzyme concentration 0.12 v/v, initial substrate concentration 0.42 mg/ml, ionic strength 0.2 M and under these optimal conditions, the maximum rate of gallic acid production was 33.49 mumoles/ml/min.