Molecular cloning of a bacterial phytase gene
2013
Humma, R.
Phytase enzyme releases myo-inositol phosphate by stepwise hydrolyses of phytic acid into myo-inositol and phosphoric acid. The efficiency of removing phytic acid by microbial fermentation using non thermophilic microorganisms is significantly decreased at high temperature in commercial point of view. For the purpose of cloning of thermostable phytase gene from thermophilic bacteria, Bacillus subtilis is selected which can bear temperature more than 50 degree C. Bacillus subtilis was grown on Phytase screening medium. Total cellular DNA has been extracted from the Bacillus subtilis. The primers were designed through appropriate software for polymerase chain reaction PCR to isolate thermostable phytase gene. PCR product of 1290 bp was confirmed on 1% agarose gel. PCR products was cloned in T/A cloning vector and analyzed by colony PCR and confirmed on 1.5% agarose gel. The industrial applications of such a phytase producing microorganism is promising as the phytase produced thereby is thermostable and fermentation using the same is efficient and energy saving.
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