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PURIFICATION, CHARACTERIZATION AND GENE ENCODING OF XYLANASE PRODUCED FROM BACILLUS TEQUILENSIS SH0 ISOLATED FROM COMPOST USING LOW COST WHEAT BRAN AS SUBSTRATE

Sanjeev Kumar, Nivedita Sharma & Shruti Pathania

A potential bacterial isolate exhibiting extracellular cellulase-free xylanase activity by Bacillus tequilensis SH0 was isolated from compost. The isolate demonstrating maximal xylanase activity after optimization using COFAT i.e. basal salt medium at 96 h, pH 5.5, temperature 45ºC, inoculums size 10%, carbon source-wheat bran (1.25%). The multistep purification techniques used were ion exchange chromatography and gel exclusion chromatography. The molecular weight was found in range of 14kDa- 97.4 kDa. The purified xylanase showed optimal activity of 41.30 IU/ml at 90ºC, pH 6.0 on xylan and also depicted cellulase free nature. The Km and Vmax of partially purified xylanase from B.tequilensis SH0 were 1.55 mg/ml and 125.0 µ mol/mg/min. Encoding of genes responsible for xylanase production was done using gradient PCR.