IMPACT OF MUTATION ON BACILLUS SP FOR THE PRODUCTION EXTRACELLULAR ALKALINE PHOSPHATASE (ALPs)
*Somashekhar R.
ABSTRACT
ALPs used as therapeutic agents and therapeutic targets show several application in clinical medicine and in biotechnology. An example used in a technique for labelling monoclonal antibodies. ALPs has the biggest market volume, to reach the market value we made an attempt to enhance the ALPs production by mutation on organisms. Six organisms were isolated using soil samples collected from different localities of Karnataka. Among the six organisms, one of the organism showed more activity based on the maximum yellow colour zone developed surrounding the organisms on the selective media and also maximum ALPs activity. This organism was identified as Bacillus sp based on the morphological, biochemical tests and 16s rRNA sequence. ALPs activity was observed maximum at 35̊C, pH 7 and 30 hrs of incubation. This organism was subjected to UV exposure at different time intervals such 5, 10, 15, 20 and 25min with different distance such 20 cm, 40 cm and 60 min at 250 nm. The result reveals that 22% enzyme activity increased at 40 cm with 15 min. In addition to this the organism was subjected X-ray with different greys such as 30, 60 and 90. Result reveals that, 6% of the enzyme activity was increased at 30 greys. After purification by Ammonium sulphate precipitation, dialysis, ion exchange chromatography and gel filtration, Fold purification was increased up to 4.02 and 40% yield. Molecular weight of the enzyme 64kDa was determined by SDS-PAGE.
Keywords: Mutation, X-ray, Ion exchange Gel filtration and SDS –PAGE.
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