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Optimization of production
of subtilisin in solid substrate fermentation using response
surface methodology
Varun Bhaskar, Jones Raj
T. R., Kandasamy S. K. J., Vijaykumar P. and Anant Achary*
Department
of Biotechnology, Kamaraj College of Engineering and
Technology, S. P. G. C. Nagar, Virudhunagar – 626 001, Tamil
Nadu, India.
*Corresponding author. E-mail:
achyanant@yahoo.com.
Tel: 0091- 4549- 278171. Fax: 0091- 4549- 278172.
Accepted
20 March, 2008 |
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Subtilisin (EC 3.4.21.62) is a type of serine protease that
is of high commercial importance. It is mainly produced by
Bacillus species as an extra cellular enzyme.
Subtilisin being stable over a wide range of pH and
temperature is exploited as a detergent enzyme and also
favoured by its non-specificity. The yield of this enzyme
needs to be optimized for cost effective production of
subtilisin for commercialization. Here, various solid
substrates were screened for the production of subtilisin
using Bacillus subtilis MTCC 441 and the maximum
yield was found with green gram husk. Response surface
methodology (RSM) was used to optimize pH, temperature, and
moisture content. The optimal conditions were obtained after
solving the polynomial equation using inverse matrix. The
optimum pH, temperature and moisture content obtained under
the conditions of study were 7.00, 30.04°C, and 73.38%
respectively. The predicted response under these conditions
was to be 571.73 U/mg protein whereas the actual response
was found to be 571.32 U/mg protein.
Key
words:
Bacillus
subtilis,
subtilisin, solid substrate fermentation, response surface
methodology, Box Benkhn Design. |
