Open AccessThermo Stability Study of Crude Amylase from Bacillus Isolate
Author(s) -
Madan Lal,
Shiv Kumar Verma,
Vijaya Nath Mishra,
Mita Chatterjee Debnath
Publication year - 2015
Publication title -
international journal of applied sciences and biotechnology
Language(s) - English
Resource type - Journals
ISSN - 2091-2609
DOI - 10.3126/ijasbt.v3i3.13219
Subject(s) - amylase , extracellular , fermentation , strain (injury) , ammonium , enzyme , laboratory flask , chemistry , yeast extract , food science , enzyme assay , ammonium sulfate precipitation , bacillus (shape) , bacillales , bacillus subtilis , chromatography , yield (engineering) , bacteria , biology , biochemistry , microbiology and biotechnology , genetics , materials science , organic chemistry , metallurgy , anatomy , size exclusion chromatography
An amylolytic strain was collected from rotten potato and its activity evaluated. The isolated strain was cultivated for amylase production in shake flasks at 35±2oC and the fermentation pattern was studied. Optimum temperature for maximum enzyme synthesis was observed at 35°C, when initial pH of fermentation medium was adjusted to 5.5. Maximum extracellular amylase activity of 7.9 U/mL and the maximum intracellular activity of 320 U/mL was recorded. Although maximum biomass was present at 12.6 g/L but highest growth rate was observed between 08 to 40h with maximum at 36h. The extracellular amylase present in the broth was partially purified with an overall yield of 44% through purification procedure of ammonium sulphate precipitation. After completed extraction and partial purification and stabilization, the stability of enzyme was observed in a range of temperature and pH between 60°C-90°C and 2-8 pH respectively. Maximum enzyme activity was demonstrated at 90°C, and pH of 5.5 and 6.5. The thermo stability of the amylases of this Bacillus species was comparable to that of amylases from other microbial sources.Int J Appl Sci Biotechnol, Vol 3(3): 427-430