Rina Rani Ray

Extracellular β-amylase from Syncephalastrum racemosum

Syncephalastrum racemosum RR 96, MTCC 2774, synthesized extra-cellular β-amylase (EC 3.2.1.2) when cultivated with starch as sole carbon source. The strain showed maximum β-amylolytic activity on the fourth day of cultivation, and the partly purified enzyme showed highest activity at 60° and pH 5. About 60% of the activity was found to be retained within the range pH 4-7. The enzyme activity was not increased in presence of exogenous thiols and remained unaffected in presence of heavy metals and the thiol inhibitor p -chloromercuribenzoate, indicating the absence of thiols at the active site of the enzyme. Absence of substrate cross specificity and ability to hydrolyse waste starches made the enzyme competent for industrial application.

A Statistical Approach for Optimization of Simultaneous Production of β-Glucosidase and Endoglucanase by Rhizopus oryzae from Solid-State Fermentation of Water Hyacinth Using Central Composite Design

The production cost of β-glucosidase and endoglucanase could be reduced by using water hyacinth, an aquatic weed, as the sole carbon source and using cost-efficient fermentation strategies like solid-state fermentation (SSF). In the present study, the effect of different production conditions on the yield of β-glucosidase and endoglucanase by Rhizopus oryzae MTCC 9642 from water hyacinth was investigated systematically using response surface methodology. A Central composite experimental design was applied to optimize the impact of three variables, namely, substrate concentration, pH, and temperature, on enzyme production. The optimal level of each parameter for maximum enzyme production by the fungus was determined. Highest activity of endoglucanase of 495 U/mL was achieved at a substrate concentration of 1.23%, pH 7.29, and temperature 29.93°C whereas maximum β-glucosidase activity of 137.32 U/ml was achieved at a substrate concentration of 1.25%, pH 6.66, and temperature 32.09°C. There was a direct correlation between the levels of enzymatic activities and the substrate concentration of water hyacinth as carbon source.

Biochemical approaches of increasing thermostability of β‐amylase from Bacillus megaterium B6

Studies on the irreversible thermoinactivation of β‐amylase from Bacillus megaterium B6 exposed to 60°C revealed that the deactivation mechanism probably results from the oxidation of thiols present at the active site of the enzyme. Several attempts were made to increase its thermostability, which indicated that Mn2+ played a key role in determining thermostability and partially reactivating the inactivated enzyme. Immobilization of β‐amylase through gel‐entrapment and covalent crosslinking brought about a remarkable increase in thermotolerance with about a 14‐fold increase in catalytic half‐life.

Extra-cellular isoamylase production by Rhizopus oryzae in solid-state fermentation of agro wastes

ABSTRACT Extra-cellular isoamylase was produced by Rhizopus oryzae PR7 in solid-state fermentations of various agro wastes, among which millet, oat, tapioca, and arum (Colocasia esculenta ) showed promising results. The highest amount of enzyme production was obtained after 72 h of growth at 28°C. The optimum pH for enzyme production was - 8.0. Among the various additives tested, enzyme production increased with ions such as Ca 2+ , Mg 2+ and also with cysteine, GSH, and DTT. The enzyme synthesis was reduced in the presence of thiol inhibitors like Cu 2+ and pCMB. The surfactants like Tween-40, Tween-80 and Triton X-100 helped in enhancing the enzyme activity. The production could be further increased by using the combinations of substrates. The ability to produce high amount of isoamylase within a relatively very short period and the capability of degrading wastes could make the strain suitable for commercial production of the enzyme. Key words: Isoamylase, Rhizopus oryzae , SSF, waste utilization

β-Amylase fromBacillus megaterium

Bacillus megaterium strain B6 producing extracellular β-amylase was isolated and grown in a medium supplemented with waste potato starch. It showed highest enzyme synthesis in the early stationary phase. The partially purified β-amylase had a temperature optimum at 60°C and a pH optimum at 6.9 and was not affected by Schardinger dextrins. These properties would allow its application in sugar industry.

Simultaneous production of animal feed enzymes (endoxylanase and endoglucanase) by Penicillium janthinellum from waste jute caddies

BackgroundIn West Bengal, India, a huge amount of lignocellulosic jute waste products, in the form of unutilized cut pieces of jute caddies consisting 65% cellulose, 22.5% hemicellulose, and 11% lignin, is generated from jute industries. Naturally, the high percentage of lignocellulolytic material made it an attractive substrate for the hydrolytic cleavage of lignocellulolytic enzymes, namely xylanases and cellulases. The present study aims for the production of xylanolytic and cellulolytic enzymes by a fungal isolate Penicillium janthinellum MTCC10889 using a cheap medium containing residual jute caddies obtained from jute mill wastes and chemically defined basal medium under submerged fermentation.ResultsThe fungal isolate P. janthinellum MTCC10889 is an outstanding producer of endoxylanase and also cellulase. A central composite design was applied to optimize its lignocellulolytic as well as cellulolytic enzyme production in submerged-state fermentation. The impact of three quantitative variables, namely pH, temperature, and substrate concentrations, on enzyme production was investigated by using a chemically defined basal medium supplemented with jute caddy as substrate. Such optimum conditions were as follows: pH 7.47, temperature of 30.35°C, and substrate concentration of 1.70% for endoxylanase, and pH 7.51, temperature of 29.69°C, and substrate concentration of 2.11% for cellulase production.ConclusionsA combination of one-factor-at-a-time approach is replaced by response surface methodology using central composite design, which resulted in 3.08- and 3.84-fold increases in the yields of endoxylanase (1,750 IU/ml) and endoglucanase (192 IU/ml), respectively. The highest endoxylanase and endoglucanase productions under optimized conditions were achieved within 48 h of growth.

Adsorption of β‐amylase from Bacillus carotarum B6 onto different native starches

β‐Amylase from Bacillus carotarum B6, a soil isolate, was readily adsorbed onto native raw starch from various sources. The absorbability was highest toward arrowroot starch and was complete within 30 min of exposure. Adsorption was remarkably affected in the presence of surfactants like Tween 80.

Induction and carbon catabolite repression of isoamylase production in Rhizopus oryzae PR7

The growth and the extracellular isoamylase production by Rhizopus oryzae PR7 MTCC 9642 were studied in a stationary culture at 28°C, with maximum isoamylase production obtained after 72 hours. Glycogen was found to be the best inducer for isoamylase synthesis, followed by maltose and dextrin. The enzyme was found to be repressed by glucose and this repression was not overcome by the addition of cGMP. The abrupt reduction in enzyme synthesis after the addition of exogenous glucose in a glycogen-induced culture medium confirmed the repressive action of glucose. An almost similar rate of repression was found to be exerted by α- and β-cyclodextrins. The inhibition of enzyme production after the addition of cycloheximide, a translation blocker, indicated the existence of de novo synthesis of the enzyme.

Effect of salicin on induction and carbon catabolite repression of endoxylanase synthesis in Penicillium janthinellum MTCC 10889

Amongst various carbon sources, xylan was found to be the sole inducer of endoxylanase production by Penicillium janthinellum MTCC 10889 in submerged cultivation. Endoxylanase synthesis by a xylan induced culture was initially repressed after a simultaneous addition of xylose, probably by the inducer exclusion mechanism, but it was resumed and achieved its highest level at a much later stage of growth (at 120 h). Xylose added after 30 h of growth cannot exert its full repressive effect. Although glucose was proved to be a more potent repressor than xylose, supplementation of salicin, an alcoholic β-glycoside containing d-glucose, with pure xylan resulted in an about 3.22 fold increase in the enzyme synthesis at 72 h followed by constant high production of the enzyme at least until the 144th h of growth. Inducing capacity of salicin in a xylan induced culture was significantly reduced when it was added after 30 h of growth. Addition of salicin and xylan help to partially overcome the repressive effect of xylose and glucose. Failure of salicin in recovering the endoxylanase synthesis in actinomycin D and cyclohexamide inhibited the xylan induced culture indicating that salicin cannot initiate the de novo synthesis of the enzyme.