Characterization Immobilization and Applications of Extracellular Protease from Bacillus sp ASASBT isolated from Termite Soil

Abstract

The present study was aimed to isolate, identify, purify, characterize, newlineimmobilize and study the applications of protease from a Bacillus sp. ASASBT newlineisolated from termite soil as a potential source of the enzyme protease thereby trying newlineto reduce the production cost. In Phase I, four different microbial sources, namely, newlineTermite Mound Soil (TMS), Organic Waste degraded Soil (OWS), Textile Effluent newlinedegraded Soil (TES) and Marine Soil (MRS) were collected at random within the newlinesouthern regions of Tamil Nadu. All the samples were serially diluted and spread on newlinenutrient agar plates. Morphologically distinct colonies were isolated from each newlinesample and were screened and assayed for the presence of the enzymes amylase, newlinecellulase, lipase and protease. The results of Phase I showed that a total of 45 newlinebacterial isolates were isolated from the four different soil samples. Eventhough, all newlinethe isolates were able to produce all the four enzymes, protease showed maximum newlineactivity. As a result, the maximum activity and zone of inhibition of protease was newlineexhibited by the isolate TMS1. In Phase II, the isolate TMS1 bacteria was identified newlineand its media components for protease production optimized. Thus, considering the newlinecolony, morphological and biochemical characteristics, it can be said that the newlinephenotypic characteristic of the selected strain TMS1 belongs to the genus Bacillus newlinesp. The 16S rRNA gene sequencing analysis showed that the isolate TMS1 was newlineBacillus sp. and therefore named as Bacillus sp. ASASBT and approximately 1.2 kb newlineof protease gene was amplified using the primers. Optimization of media components newlinefor protease production showed higher activity of the enzyme for an incubation period newlineof 48 hours at pH 7.0, temperature 40ºC, starch as the carbon source, gelatin as the newlinenitrogen source, green gram husk as a natural substrate and an agitation speed of 100 newlinerpm.