Exo-polygalacturonase production by Bacillus subtilis CM5 in solid state fermentation using cassava bagasse / Produção de exo-poligalacturonase por Bacillus subtilis CM5 por fermentação em estado sólido empregando bagaço de mandioca

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 percent 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 por cento e 50ºC, respectivamente.

Statistical optimization of alpha-amylase production with immobilized cells of Streptomyces erumpens MTCC 7317 in Luffa cylindrica L. sponge discs.

The purpose of this investigation was to study the effect of Streptomyces erumpens cells immobilized in various matrices, i.e., agar-agar, polyacrylamide, and luffa (Luffa cylindrica L.) sponge for production of alpha-amylase. Luffa sponge was found to be 21% and 51% more effective in enzyme yield than agar-agar and polyacrylamide, respectively. Response surface methodology was used to evaluate the effect of three main variables, i.e., incubation period, pH, and temperature on enzyme production with immobilized luffa cells. The experimental results showed that the optimum incubation period, pH, and temperature were 36h, 6.0, and 50 degrees C, respectively. The repeated batch fermentation of immobilized cells in shake flasks showed that S. erumpens cells were more or less equally physiologically active on the support even after three cycles of fermentation (3,830-3,575 units). The application of S. erumpens crude enzyme in liquefying cassava starch was studied. The maximum hydrolysis of cassava starch (85%) was obtained with the application of 4ml (15,200 units) of crude enzyme after 5 h of incubation.

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 50°C, respectively for optimum exo-PG production.

Solid substrate fermentation of cassava fibrous residue for production of alpha-amylase, lactic acid and ethanol.

There is serious concern about the disposal of solid residues left after large scale extraction of starch from cassava. Owing to the high starch content (55-65% on dry weight basis) and organic matter of these wastes, an attempt has been made to utilize it for the production of three bioproducts, i.e. alpha-amylase, lactic acid and ethanol in solid substrate fermentation by incubating the solid residue at different moisture holding capacity (40-80%) and incubation period (12- 60 hr for alpha-amylase, 24-144 hr for ethanol and 2-10 days for lactic acid). The highest product yield was obtained at 60% moisture holding capacity of the residue and period of incubation varied from 36 hr (alpha-amylase), 120 hr (ethanol) to 6 days (lactic acid). This study showed that the solid residues from cassava starch factories could serve as a low-cost substrate for bioproducts production.

Statistical optimization of alpha-amylase production by Streptomyces erumpens MTCC 7317 cells in calcium alginate beads using response surface methodology.

Alpha-amylase has a wide range of applications in starch industries, i.e. baking, brewing, distillery, etc. The alpha-amylase production from Streptomyces erumpens MTCC 7317 immobilized cells was compared with that of free cells. The immobilized cells of S. erumpens in calcium alginate beads were more effective for production of alpha-amylase (12.2% more yield) than free cells. Response surface methodology (RSM) was used to evaluate the effect of main variables, i.e. incubation period, pH and temperature on enzyme production with immobilized cells. A full factorial Central Composite Design (CCD) was applied to study these main factors that affected alpha-amylase production. The experimental results showed that the optimum incubation period, pH and temperature were 36 h, 6.0 and 50 degrees C, respectively for immobilized cells. Repeated batch fermentation of immobilized cells in shake flasks carried out in starch-beef extract medium showed that S. erumpens cells were physiologically active on the support even after four cycles of fermentation.

Statistical optimization of alpha-amylase production by probiotic Lactobacillus plantarum MTCC 1407 in submerged fermentation.

Production and purification of alpha-amylase by probiotic Lactobacillus plantarum MTCC 1407 has been investigated under submerged fermentation using Mann Rogassa Sharpe medium containing (1%) soluble starch in lieu of glucose (2%) as carbon source. Response Surface Methodology was used to evaluate the effect of main variables, i.e. incubation period, pH and temperature on enzyme production. A full factorial Central Composite Design was applied to study these main factors that affected alpha-amylase production. The experimental results showed that the optimum incubation period, pH and temperature were 36 h, 7.0 and 35 degrees C, respectively. The purified enzyme (by ammonium sulphate precipitation) had a molecular mass of 75 450 Da in SDS-PAGE.

Alpha-amylase production by Streptomyces erumpens MTCC 7317 in solid state fermentation using response surface methodology (RSM).

Production of alpha-amylase under solid state fermentation by Streptomyces erumpens MTCC 7317 has been investigated using different agro-industrial residues, i.e. cassava bagasse, sugarcane bagasse and wheat bran; wheat bran was found to be the best substrate. Among different nitrogen source supplemented to wheat bran, beef extract or peptone (1%) showed maximum enzyme production. Response surface methodology was used to evaluate the effect of main process parameters as incubation period (48 h), moisture holding capacity (70%), pH (7.0) and temperature (50 degrees C) on enzyme production by applying a full factorial central composite design. The maximum hydrolysis of soluble starch (90%) and cassava starch (75%) was obtained with the application of 4 ml (approximately 12096 U) of S. erumpens crude enzyme after 5 h of incubation.

Partial characterization and optimization of production of extracellular alpha-amylase from Bacillus subtilis isolated from culturable cow dung microflora.

Studies of alpha-amylase production by Bacillus subtilis (CM3) isolated earlier from cow dung microflora, were carried out. The optimum temperature, pH and incubation period for amylase production were 50-70 degrees C, 5.0-9.0 and 36 h, respectively. Enzyme secretion was very similar in the presence of any of the carbon sources tested (soluble starch, potato starch, cassava starch, wheat flour, glucose, fructose, etc.). Yeast extract and ammonium acetate (1%) as nitrogen sources gave higher yield compared to other nitrogen sources (peptone, malt extract, casein, asparagine, glycine, beef extract), whereas ammonium chloride, ammonium sulphate and urea inhibited the enzyme activity. Addition of Ca+2 (10-40 mM) to the culture medium did not result in further improvement of enzyme production, whereas the addition of surfactants (Tween 20, Tween 40, Tween 80, and sodium lauryl sulphate) at 0.02% resulted in 2-15% increase in enzyme production. There were no significant variations in enzyme yield between shaked-flask and laboratory fermentor cultures. The purified enzyme is in two forms with molecular mass of 18.0 +/- 1 and 43.0 +/- 1 kDa in native SDS-PAGE.