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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology

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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.

New Biotechnology

|June 15, 2010

Summary

Researchers optimized laccase production using solid state fermentation (SSF) from Pleurotus sp. This resulted in the highest reported laccase titer, demonstrating the enzyme

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Area of Science:

Biotechnology
Enzymology
Microbial Fermentation

Background:

Laccases are versatile enzymes with broad industrial applications.
Optimizing laccase production is crucial for cost-effective industrial use.
Pleurotus sp. is a known source of laccase, but strain-specific optimization is needed.

Purpose of the Study:

To optimize laccase production from a hyperactive Pleurotus sp. strain using solid state fermentation (SSF).
To investigate the interactions between fermentation parameters and their effect on laccase yield.
To characterize the laccase isoforms produced and their properties.

Main Methods:

Solid state fermentation (SSF) using a locally isolated Pleurotus sp. strain.
Response surface methodology (RSM) to study parameter interactions.
Isoelectric focusing (IEF) to analyze enzyme isoforms.
Differential evolution technique coupled with regression analysis for optimization.

Main Results:

Identified dual production optima for laccase, suggesting the presence of isozymes.
Two laccase isoforms with distinct pI values (3.8 and 9.3) were detected.
Optimized conditions yielded a record laccase titer of 54,600 IU/gds (3,412,500 U/L).
Optimal conditions included specific surfactant concentration, pH, particle size, liquid-to-solid ratio, and incubation period.

Conclusions:

The study successfully optimized laccase production from Pleurotus sp. using SSF.
The identified laccase isoforms exhibit potential for applications in both acidic and alkaline environments.
The achieved laccase yield represents a significant advancement in enzyme production.