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Journal of Microbial & Biochemical Technology
Catalytic oxidation and enhanced production of oxidoreductases for polymer degradation
5th International Conference on Microbial Physiology and Genomics
September 29-30, 2016 London, UK

Muhammad Ishtiaq Ali and Nazia Khatoon

Quaid-i-Azam University, Pakistan

Posters & Accepted Abstracts: J Microb Biochem Technol

Abstract:

Oxidative enzymes play significant role in biodegradation of recalcitrant materials. Fungi are important among microorganisms for production of extracellular enzymes. Limited production and slow release of the particular enzymes are the limiting factor. The present study aimed for enhanced production, molecular characterization of oxidoreductases. Molecular examination as well as the heterologous expression of ligninolytic enzymes i.e., laccase and lignin peroxidase were carried out. These enzymes are mainly produced under nutrient starved condition i.e., carbon or nitrogen limited medium. Microscopic examination of these enzymes producing organism showed that they are filamentous, coenocytic, aseptate and spore producing organisms. An experiment was set up by adding the PVC polymer in the MSM media and inoculating the respective enzymes after screening and purification. The Fourier transform infrared (FTIR) spectroscopy of enzyme treated plastic films revealed the structural changes as compared to control (without enzyme treatment. Enzyme assay of both enzymes such as laccase and lignin peroxidase were carried out with vertryl alcohol and DMP as substrates. The extracted DNA fragments of both enzymes were then amplified by PCR with Lip and Lac primers. The amplified PCR product 530 bp and 890 bp then ligated with pTZ57R/T plasmid. Competent cells were developed through both chemical and electroporation. The competent cells used for the expression purpose were the DHα5 strain of E. coli bacterium. Confirmation of the cloned fragment was done through restriction endonuclease enzymes Eco R1 and Hind111.

Biography :

Email: ishi_ali@hotmail.com