NDLI: Expression and characterization of Trichoderma virens UKM-1 endochitinase in Escherichia coli

Content Provider	Springer Nature Link
Author	Alias, Nadiawati Mahadi, r Muhammad Murad, Abdul Munir Abdul Bakar, Farah Diba Abu Mahmood, Nik Azmi Nik Illias, Rosli Md
Copyright Year	2008
Abstract	A gene encoding endochitinase from Trichoderma virens UKM-1 was cloned and expressed in E. coli BL21 (DE3). Both the endochitinase gene and its cDNA sequences were obtained. The endochitinase gene encodes 430 amino acids from an open reading frame comprising of 1,690 bp nucleotide sequence with three introns. The endochitinase was expressed as soluble and active enzyme at 20°C when induced with 1 mM IPTG. Maximum activity was observed at 4 h of post-induction time. SDS-PAGE showed that the purified endochitinase exhibited a single band with molecular weight of 42 kDa. Biochemical characterization of the enzyme displayed a near neutral pH characteristic with an optimum pH at 6.0 and optimum temperature at 50°C. The enzyme is stable between pH 3.0–7.0 and is able to retain its activity from 30 to 60°C. The presence of Mg$^{2+}$ and Ca$^{2+}$ ions increased the enzyme activity up to 20%. The purified enzyme has a strong affinity towards colloidal chitin and low effect on ethyl cellulose and D-cellubiose which are non-chitin related substrates. HPLC analysis from the chitin hydrolysis showed the release of (GlcNAc)$_{3}$, (GlcNAc)$_{2}$ and GlcNAc, in which (GlcNAc)$_{2}$ was the main product.
Starting Page	561
Ending Page	572
Page Count	12
File Format	PDF
ISSN	09593993
Journal	World Journal of Microbiology and Biotechnology
Volume Number	25
Issue Number	4
e-ISSN	15730972
Language	English
Publisher	Springer Netherlands
Publisher Date	2008-12-13
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Trichoderma virens Recombinant chitinase Expression Inclusion bodies Characterization Microbiology Environmental Engineering/Biotechnology Applied Microbiology Biochemistry Biotechnology
Content Type	Text
Resource Type	Article
Subject	Physiology Medicine Applied Microbiology and Biotechnology Biotechnology

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