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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
Optimization of laccase production using response surface methodology coupled with differential evolution
Sukanta Shekhar Bhattacharya1, Vijay Kumar Garlapati, Rintu Banerjee
1Microbial Biotechnology and Downstream Processing Laboratory, Department of Agricultural and Food Engineering, Indian Institute of Technology, Kharagpur, Kharagpur, West Bengal 721 302, India.
Abstract:
In the present study, laccase production from a locally isolated hyperactive strain of Pleurotus sp. under solid state fermentation (SSF) was carried out and the interactions between different parameters of fermentation were studied using response surface methodology. The saddle shaped response surface plots depicting dual conditions for the enhanced production indicated the presence of isozymes with production optima at different conditions which was verified experimentally. Isoelectric focusing of the enzyme extract revealed that two isoforms were found with a widely varying pI of 3.8 and 9.3 emphasizing the capacity of the enzyme to be deployed at both acidic and alkaline conditions. Optimization of production conditions by coupling the regression equation with differential evolution technique yielded over 54,600IU/gds (3,412,500U/L) with a surfactant concentration of 0.016%, pH 7.99, particle size of 0.25cm, liquid to solid ratio of 4.99 and an incubation period of 8 days. In this study, the optimization process yielded highest titer value of laccase reported to date.
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Response Surface Methodology
The process of RSM involves several key steps:
