Overexpression of a Chitinase Gene from the Thermophilic Fungus, Thermomyces lanuginosus in Saccharomyces cerevisiae and Characterization of the Recombinant Chitinase
M. Prasad and P. Palanivelu*
Department of Molecular Microbiology, School of Biotechnology, Madurai Kamaraj University, Madurai, India
- *Corresponding Author:
- P. Palanivelu
Department of Molecular Microbiology
School of Biotechnology
Madurai Kamaraj University, Madurai, India
Tel: 0452-2458208, 9486209859
E-mail: [email protected]/ [email protected]
Received Date: November 08, 2012; Accepted Date: December 17, 2012; Published Date: December 19, 2012
Citation: Prasad M, Palanivelu P (2012) Overexpression of a Chitinase Gene from the Thermophilic Fungus, Thermomyces lanuginosus in Saccharomyces cerevisiae and Characterization of the Recombinant Chitinase. J Microb Biochem Technol 4: 086-091. doi: 10.4172/1948-5948.1000076
Copyright: © 2012 Prasad M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
A chitinase gene from the thermophilic fungus, Thermomyces lanuginosus ATCC 44008 has been cloned and overexpressed in Saccharomyces cerevisiae SEY 2101. The recombinant chitinase was produced as soluble secreted protein. The enzyme activity was found to be maximum on fourth day at 30°C in the induction medium. The overexpressed chitinase displayed optimum activity at pH 6.5 and at 60°C. The recombinant chitinase attributed remarkable thermostability by holding more than 60% of the enzyme activity after 6 h at 50°C. The molecular mass of the overexpressed chitinase was 42 kDa as measured by SDS-PAGE. The kinetic parameters such as KM and Vmax of the enzyme were found to be 0.403 mM and 8.74 mmoles/min/mg protein, respectively. Synthesis of the recombinant chitinase was strongly repressed by glucose. The eukaryotic translational inhibitor, cycloheximide in the induction medium showed 10% of higher activity whereas 30% of the activity was inhibited by transcriptional inhibitors, viz. 8-azaguanine and 8-hydroxyquinoline. The overexpressed recombinant chitinase may find potential application in pharmaceutical industry to prepare chitooligosaccharides.