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Production and Partial Purification of Alpha Amylase from Bacillus subtilis (MTCC 121) Using Solid State Fermentation
Dibyangana Raul1, Tania Biswas1, Suchita Mukhopadhyay1
1Department of Biotechnology, Haldia Institute of Technology, ICARE Complex, Purba Medinipur 721657, India.
Biochemistry Research International
|March 28, 2014
Summary
Solid state fermentation using Bacillus subtilis produced high yields of alpha-amylase enzyme. This enzyme, crucial for starch breakdown, showed optimal activity at pH 7.1 and 40°C after 48 hours of fermentation.
Area of Science:
- Biochemistry
- Enzymology
- Industrial Microbiology
Background:
- Amylase is a vital enzyme for starch hydrolysis, with applications in food, biofuel, and industrial processes.
- Solid-state fermentation (SSF) offers advantages over submerged fermentation (SmF) for enzyme production, including simplicity and cost-effectiveness.
Purpose of the Study:
- To optimize alpha-amylase production using Bacillus subtilis via solid-state fermentation.
- To characterize the optimal conditions and partially purify the produced alpha-amylase.
Main Methods:
- Solid-state fermentation (SSF) of Bacillus subtilis on wheat bran for 48 hours at 37°C.
- Enzyme activity assay (quantitative DNS assay) and zymogram analysis.
- Partial purification using ammonium sulfate precipitation ((NH4)2SO4).
Main Results:
- Maximum alpha-amylase yield was achieved after 48 hours of SSF.
- The enzyme exhibited optimal activity at pH 7.1 and 40°C.
- Amylase activity was concentrated threefold using a 30-70% (NH4)2SO4 precipitation cut.
Conclusions:
- Solid-state fermentation is an effective method for producing alpha-amylase from Bacillus subtilis.
- The characterized alpha-amylase shows potential for various industrial applications.
- Further purification is required to determine the enzyme's precise molecular weight.
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