Gryllotalpa Case Study

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An insect gut of Gryllotalpa was isolated, identified as a tannase producing bacterial strain as E. cloacae on the basis of its different characteristics morphological, physiological and biochemical (Beniwal et al., 2010). Enzyme production was carryout in a 250 ml Erlenmeyer flask containing 100 ml of basal medium [(g/l) KH2PO4, 0.5; CaCl2, 0.5; MgCl2, 2.0; NaNO3, 2.0] supplemented with 12 g/l tannic acid as a sole carbon source. The medium was inoculated with 1% of 18 h old inoculums culture were incubated at 37°C for 24 h at an agitation speed of 100 rpm. Tannase enzyme assay and protein estimation of E. cloacae Tannase activities were determined spectrophotometrically by the method of Mondal et al. (2001). One unit of the tannase was defined as the amount of enzymes,…show more content…
Km and Vmax values were calculated for the substrate. Effect on reaction time The tannase enzyme sample and the substrate tannic acid were incubated for 15, 30, 45, 60, 75 and 90 mins. Standard protocol was used for tannase assay. Salt tolerance test Effect of NaCl on purified tannase was determined at different concentrations of NaCl ranging from (1–5M) Salt tolerance of the purified tannase was determined by incubating the enzyme at different concentrations of NaCl, up to 5 M for 1 h incubation at 50°C. The residual activity was calculated under the standard assay conditions. Effect of inhibitors The enzyme solution containing 1 mM concentration of various inhibitors like Gallic acid, Methyl gallate, Pyrogallol, n-Propyl gallate, Sodium bisulfate, and Sodium thioglycolate was incubated at 50°C for duration of time 5 and 60 min. The enzyme samples were then assayed for tannase enzyme activity. Effect of chelators and surfactants on tannase

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