Isolation of a kojic acid-producing fungus capable of using starch as a carbon source

A fungal strain (S33-2), able to grow on cooked starch and produce a substantially high level of kojic acid, was isolated from morning glory flower (Bixa orellana). The fungus was characterized and identified as Aspergillus flavus. The effect of different types of starch (sago, potato and corn starch) on growth of strain S33-2 and kojic acid production was examined using shake flasks. It was found that strain S33-2 grew well on all types of starch investigated. However, kojic acid production was highest when corn starch was used, with the maximum kojic acid obtained being comparable to fermentation using glucose. The highest kojic acid production (19.2 g l-1) was obtained when 75 g l-1 corn starch was used. This gave a yield, based on starch consumed, and an overall productivity of 0.256 g g-1 and 0.04 g l-1 h-1, respectively

The effect of carbon sources on the expression level of thermostable L2 lipase in Pichia pastoris

Thermostable lipases are enzymes that are particularly pleasing for industrial purposes such as in the production of detergents, animal skin-based industry and food processing endeavours. Thermostable L2 lipase gene fished out from Bacillus sp. L2 was cloned into Pichia pastoris strain GS115 under constitutive expression system of pGAPZαA. Study performed on various carbon sources revealed that glucose and glycerol support the growth of Pichia pastoris and expression of L2 lipase. In addition, the by-product of sugar refinery processes (molasses) was found to be a potential carbon source for the growth of P. pastoris and L2 lipase expression. Since the thermostable L2 lipase will be extracellularly secreted into the medium, the use of P. pastoris expression system is seen as an alternative to the conventional expression system of Escherichia coli that applies intracellular expression of the L2 lipase gene encoded.Key words: Thermostable lipase, Pichia pastoris, constitutive expression, carbon source

Effects of Aspergillus niger (K8) on nutritive value of rice straw

The objective of this study was to evaluate the use of solid state fermentation for the improvement of the quality of rice straw as animal feed. Rice straw was fermented using Aspergillus niger (K8) with and without additional nitrogen source (urea). Cellulose, hemicelluloses, organic matter (OM), dry matter (DM), acid detergent fibre (ADF), neutral detergent fibre (NDF) and acid detergent lignin (ADL) contents of rice straw were determined before and after 10 days of fermentation. Fermentation has significant (P < 0.01) effect on NDF, but not ADF and ADL contents. Addition of urea as nitrogen source significantly reduced (P < 0.01) the NDF and hemicellulose contents of fermented rice straw. Cellulose content of the rice straw was not affected (P > 0.05), but crude protein (CP) increased significantly (P < 0.01) after fermentation. In vitro gas production technique was used to evaluate the effect of the biological treatment on activity of rumen microorganisms. Fermentation of rice straw using A. niger significantly reduced total gas production (P < 0.01), DM disappearance (P < 0.01) and acetate, propionate and  total volatile fatty acids (VFA) production (P < 0.05). Results of the present study showed that solid state fermentation of rice straw using A. niger reduced lignocellulose content, but has negative effect on microbial activity in the rumen ecosystem, presumably due to antagonistic activity of A. niger, or other intermediate products from the fermentation, on the rumen microorganisms.Key words: Aspergillus niger, biomass, solid state fermentation, biological treatment, in vitro gas production

Efficiency of rice straw lignocelluloses degradability by Aspergillus terreus ATCC 74135 in solid state fermentation

The ability of Aspergillus terreus for the production of cellulolytic enzymes and reduction of lignocellulose contents of rice straw in solid state fermentation was investigated in this study. Results suggested that, 8 days fermentation was appropriate, with enzymes activities as follows: FPase = 410.76 U/gDM, CMCase = 351.96U/gDM, -glucosidase = 16.37 U/gDM, xylanase = 6166.01 U/gDM and amyloglucosidase = 425.04 U/gDM (with maximum 993.71 U/gDM on day 6). In addition, the solid state fermentation significantly (P < 0.01) reduced the concentrations of NDF, ADF, cellulose and hemicellulose in the rice straw by 19.96, 13.8, 16.32 and 32.87%, respectively. The high degradation of the hemicellulose was reflected by the high activity of xylanase enzyme, which hydrolyses xylan in hemicellulose to xylose. Higher reducing sugar and microbial cell mass productions were also obtained after 8 days fermentation. Present data showed that, A. terreus is capable of producing high quantity of cellulolytic enzymes for the reduction of lignocellulose contents of biomass in a shorter incubation time when compared with the previously reported for biological treatment of agricultural by-products using white rot fungi.Key words: Aspergillus terreus, biomass, biological treatment, enzyme activity, solid state fermentation

Extracellular Protein Secreted by Bacillus subtilis ATCC21332 in the Presence of Streptomycin Sulfate

The extracellular proteins secreted by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was carried out to determine the effect of Streptomycin Sulfate in regulating extracellular proteins secreted by Bacillus subtilis ATCC21332. Results of Microdilution assay showed that the Minimum Inhibition Concentration (MIC) of Streptomycin Sulfate on B. subtilis ATCC21332 was 2.5 mg/ml. The bacteria cells were then exposed to Streptomycin Sulfate at concentration of 0.01 MIC before being further incubated for 48h to 72 h. The extracellular proteins secreted were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile revealed that three additional bands with approximate sizes of 30 kDa, 22 kDa and 23 kDa were appeared for the treated bacteria with Streptomycin Sulfate. Thus, B. subtilis ATCC21332 in stressful condition with the presence of Streptomycin Sulfate at low concentration could induce the extracellular proteins secretion

Extracellular Protein Secreted by Bacillus subtilis ATCC21332 in the Presence of Streptomycin Sulfate

The extracellular proteins secreted by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was carried out to determine the effect of Streptomycin Sulfate in regulating extracellular proteins secreted by Bacillus subtilis ATCC21332. Results of Microdilution assay showed that the Minimum Inhibition Concentration (MIC) of Streptomycin Sulfate on B. subtilis ATCC21332 was 2.5 mg/ml. The bacteria cells were then exposed to Streptomycin Sulfate at concentration of 0.01 MIC before being further incubated for 48h to 72 h. The extracellular proteins secreted were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile revealed that three additional bands with approximate sizes of 30 kDa, 22 kDa and 23 kDa were appeared for the treated bacteria with Streptomycin Sulfate. Thus, B. subtilis ATCC21332 in stressful condition with the presence of Streptomycin Sulfate at low concentration could induce the extracellular proteins secretion

Comparative analyses on medium optimization using one-factor-at-a-time, response surface methodology, and artificial neural network for lysine–methionine biosynthesis by Pediococcus pentosaceus RF-1

Optimization strategy that encompassed one-factor-at-a-time (OFAT), response surface methodology (RSM), and artificial neural network method was implemented during medium formulation with specific aim for lysine-methionine biosynthesis employing a newly isolated strain of Pediococcus pentosaceus RF-1. OFAT technique was used in the preliminary screening of factors (molasses, nitrogen sources, fish meal, glutamic acid and initial medium pH) before proceeded to optimization study. Implementation of central composite design of experiment subsequently generated 30 experimental runs based on four factors (molasses, fish meal, glutamic acid, and initial medium pH). From RSM analysis, a quadratic polynomial model can be devoted to describing the relationship between various medium components and responses. It also suggested that using molasses (9.86 g/L), fish meal (10.06 g/L), glutamic acid (0.91 g/L), and initial medium pH (5.30) would enhance the biosynthesis of lysine (15.77 g/L) and methionine (4.21 g/L). Alternatively, a three-layer neural network topography at 4-5-2 predicted a further improvement in the biosynthesis of lysine (16.52 g/L) and methionine (4.53 g/L) by using formulation composed of molasses (10.02 g/L), fish meal (18.00 g/L), and glutamic acid (1.17 g/L) with initial medium pH (4.26), respectively