L-Glutaminase, an amidohydrolase enzyme has been a choice of interest in the treatment of lymphoblastic leukaemia. The present study reports production, purification and characterization of extracellular glutaminase enzyme from Actinomycetes. Screening was performed for twenty Actinomycetes isolates from soil; one isolate (Isolate 2) was finally selected based on the activity of glutaminase (32.5 U/ml).The isolate was identified as Streptomyces sp. Effect of physicochemical factors namely temperature, pH, NaCl concentration, and supplementary carbon & nitrogen sources on the production of L-glutaminase from the Streptomyces sp. was carried out. The enzyme production was found to be optimum with glucose as carbon source (33 U/ml), Lglutamine as nitrogen source (33.1 U/ml), at 7 pH (32.8 U/ml), temperature 30 o C (32.4 U/ml) and for 0.1% NaCl concentration (32.5 U/ml). The L-glutaminase produced from Streptomyces sp. was purified by ammonium sulphate precipitation, dialysis method and ion exchange chromatography. After the purification of the enzyme by ion exchange chromatography, it has been purified 46-fold from cell-free extract and yield was 3.25%. Characterization of extracellular L-glutaminase showed that the enzyme shown optimal activity at temperature of 30°C, pH 7, at 2% NaCl and for 0.04M substrate and the Km value was calculated to be 2.8mM and Vmax was 7.57 U/ml. The molecular weight of enzyme as determined by sodium dodecyl sulphate polyacrylamide electrophoresis (SDS-PAGE) was found to be 50 kDa.
Isolation of two keratinolytic bacterial strains from poultry soil as well as purification and properties of keratinase were investigated. Isolates were designated as KI8101 and KI8102 (KI, keratin isolates) and were identified as Bacillus subtilis and B. licheniformis respectively. The purified enzyme from KI8102 exhibited a high specific activity of 500 U/mg with 71‐fold purification and 41% yield. SDS‐PAGE analysis indicated that the purified keratinase had a molecular mass of 32 kDa. The optimum temperature and pH were 50°C and 7.5, respectively. Its Km was 83.3 μM and Vmax was 71.4 μmol/mL min. The bacterium could potentially degrade keratin waste such as human hair, nails, bovine hair and wool. Therefore, the enzyme could improve the nutritional value of meat and poultry‐processing waste containing keratin and could be a potential candidate for biotechnological processing involving keratin hydrolysis.
No abstract
Dairy Industries produce nearly thousands of litres of effluent waste per day. This waste with high intense foul odour pollutes ecosystem and ground water, harbour pathogens causing health hazards. Various pre-treatments methods are available to neutralise the effects. But, bioremediation with EM technology is ecofriendly and helps to clean up contaminated environments through the use of microorganisms.EM technology meaning Effective Microorganisms consisting of beneficial and highly efficient microbes that are non-harmful, non-pathogenic, not-genetically-engineered or modified (non-GMO), and not-chemically-synthesized. This is proven safe, lowcost, effective and easily utilized in environmental protection.In the present study, dairy industry effluent was analysed for microbial content and five different microbes were isolated and labelled as MB1, MB2, MB3, MB4 and MB5. All these were Gram negative, rod shaped and motile. The MB1 colonies displayed Green fluorescence when exposed to UV light; hence it was completely characterized and revealed to be Pseudomonas aureginosa. Biochemical analysis for reducing sugar, protein, lipids was carried out. Also BOD, COD was analysed. Qualitative analysis of Lipids was done by TLC using sprays of ninhydrin solution and molybdic acid which indicated the presence of cholesterol and phospholipids respectively. Quantitative analysis of reducing sugars by Anthrone's test and proteins by Lowry's method gave a result of total carbohydrate content of 500µg/ml and protein content of 100µg/ml. EM mixture used for effluent treatment included Lactobacillus acidophilus, Chromatium Species, Mucor heimalis, Streptomyces greiceus, Aspergillus Oryzae, Yeast, and Pseudomonas isolated from effluent tank. All the organisms were grown on different media and treated directly on 100 ml of effluent and studied for changes in three parameters viz., pH range 3-4, odour reduction to 1 in the scale of 1-5 [ 1-least/no odour, 5 -foul odour] and clearance of effluent. Except Chromatium sp., others were selected since there was no much change in parameters when Chromatium sp was used. For better results Jaggery solution was used to ferment all the organisms. Sample of effluent used was 400 ml. Odour reduction was observed after 15 days of treatment.For cost optimization sugarcane juice was used as substrate for fermentation and few more organisms were tried along with the existing EM Mixture. The organisms included lactic acid producing bacteria, Rhodopseudomonas sps and methanotroph. Sugarcane juice was fermented with different combinations of these organisms and treated on 250 ml of effluent sample. Totally 52 different combinations of EM mixture were tried, out of which 4 effective EM combinations (EM1, EM2, EM3 and EM4) were identified based on three parameters. These 4 EM mixtures were studied for scaled up effluent treatment (1litre) and it was observed that EM1 and EM2 were found to be effective in forming higher clearance in effluent turbidity with reduction in foul odour and pH , there by renderin...
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