Abstract
Amylases are one of the most important enzymes in present-day biotechnology. The present study was concerned with the production and partial characterization of extracellular amylase from Bacillus amyloliquefaciens P-001. The effect of various fermentation conditions on amylase production through shake-flask culture was investigated. Enzyme production was induced by a variety of starchy substrate but corn flour was found to be a suitable natural source for maximum production. Tryptone and ammonium nitrate (0.2%) as nitrogen sources gave higher yield compared to other nitrogen sources. Maximum enzyme production was obtained after 48 hrs of incubation in a fermentation medium with initial pH 9.0 at 42°C under continuous agitation at 150 rpm. The size of inoculum was also optimized which was found to be 1% (v/v). Enzyme production was 2.43 times higher after optimizing the production conditions as compared to the basal media. Studies on crude amylase revealed that optimum pH, temperature and reaction time of enzyme activity was 6.5, 60°C and 40 minutes respectively. About 73% of the activity retained after heating the crude enzyme solution at 50°C for 30 min. The enzyme was activated by Ca2+ (relative activity 146.25%). It was strongly inhibited by Mn2+, Zn2+ and Cu2+, but less affected by Mg2+ and Fe2+.
Highlights
Microbial enzymes are widely used in industrial processes due to their low cost, large productivity, chemical stability, environmental protection, plasticity and vast availability (Burhan et al 2003; Mishra & Behera 2008)
The optimum enzyme production by the bacterial isolate was found at 42°C, whereas maximum enzyme activity was observed at 60°C
Present study deals with the production condition optimization and partial characterization of crude extracellular amylase produced by Bacillus amyloliquefaciens P-001
Summary
Microbial enzymes are widely used in industrial processes due to their low cost, large productivity, chemical stability, environmental protection, plasticity and vast availability (Burhan et al 2003; Mishra & Behera 2008). Bacillus species such as Bacillus subtilis, Bacillus amyloliquefaciens and Bacillus licheniformis are used as bacterial workhorses in industrial microbial cultivations for the production of a variety of enzymes as well as fine biochemicals for decades. The estimated value of world market is presently about US$ 2.7 billion and is estimated to increase by 4% annually through 2012. For the maximum enzyme production, medium optimization is a prime step for its commercial usage
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