Production and partial characterization of extracellular amylase enzyme from Bacillus amyloliquefaciens P-001

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+.

Molecular Identification of Newly Isolated Bacillus Strains from Poultry Farm and Optimization of Process Parameters for Enhanced Production of Extracellular Amylase Using OFAT Method

“…A decline in amylase activity afterwards was probably due to depletion of nutrients in the fermentation medium of the microorganisms, causing stressed and unfavourable conditions for the bacteria resulting in reduction of enzyme activity. Our reports were against the findings of Vijayalakshmi et al (2012) and Deb et al (2013), who demonstrated maximum amylase activity by Bacillus sp. at 48 h of incubation.…”

“…is an industrial important microorganism because of its rapid growth rate, secretion of enzyme into the extracellular medium and safe handling (Vijayalakshmi et al, 2012). 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 (Deb et al, 2013).…”

Isolation, Characterization and Optimization of Amylase Producing Micro-Organism from Gastrointestinal Tract of Catla Catla

“…Similarly, invarious Bacillus spp. maximum amylase production was observed in pH range of 6.5 to 7.5 [33]. Earlier studies have revealed that fungi required slightly acidic pH and bacteria required neutral pH for optimum growth.…”

Isolation, Identification and Optimization of Alkaline Amylase Production from Bacillus cereus Using Agro-Industrial Wastes

“…B. cereus strain isolated from Cuddalore harbour waters grew maximally in shake flask with highest Amylase activity at pH 8.0 which was sustained even at pH 11.0 (Annamalai et al, 2011). Deb et al 2013 obtained maximum amylase production after 48 hours of incubation in fermentation medium with initial pH 9.…”

“…The result showed that, in SSF among the three substrates tested addition of Wheat Bran as substrate showed highest enzyme production of 154.2±0.65 U/ml with specific activity of 0.29±0.01 U/mg ( Figure 3A). Deb et al, 2013 showed that the production of enzyme was highest when corn flour was used as carbon source. Whereas Singh et al, (2010) found that wheat bran as suitable source for improved amylase production.…”