Publication: Biochemical characterisation of two forms of halo- and thermo-tolerant chitinase C of Salinivibrio costicola expressed in Escherichia coli
2
Issued Date
2007-01-01
Resource Type
ISSN
15904261
Other identifier(s)
2-s2.0-34547144722
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
Annals of Microbiology. Vol.57, No.2 (2007), 249-257
Suggested Citation
Ratchaneewan Aunpad, David W. Rice, Svetalana Sedelnikova, Watanalai Panbangred Biochemical characterisation of two forms of halo- and thermo-tolerant chitinase C of Salinivibrio costicola expressed in Escherichia coli. Annals of Microbiology. Vol.57, No.2 (2007), 249-257. doi:10.1007/BF03175215 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/24595
Research Projects
Organizational Units
Authors
Journal Issue
Thesis
Title
Biochemical characterisation of two forms of halo- and thermo-tolerant chitinase C of Salinivibrio costicola expressed in Escherichia coli
Other Contributor(s)
Abstract
Two forms of chitinase C (Chi-I and Chi-II) were purified until homogeneity from the culture supernatant of a transformant Escherichia coli harbouring chitinase C gene from the halophilic bacterium Salinivibrio costicola strain 5SM-1. Chi-II was derived from Chi-I by C-terminal processing. Chi-I and Chi-II showed similar salinity optimum at 1-2% NaCl and retained more than 80% of their activity at 3-5% NaCl and more than 50% residual activity at 14% NaCI. The two enzymes could also well function (activity > 95%) in the absence of NaCI. Both had highest activity at pH 7.0 and 50°C and both were stable over a wide range of pH (3.0-10.0). More than 50% activity remained at 80°C after 60 min treatment. Among 4 major cations contained in sea water, only Mg2+at 10 mM increased activity about 10%. Using ρ-nitrophenyl-N, N′-diacetylchitobiose as substrate, Chi-I and Chi-II had Kmof 30 and 31.8 μM and Vmaxof 10 and 9.2 pmol/h/mg protein, respectively. Chi-I and Chi-II were classified as exochitinases by product analysis of the E. coli culture supernatant with high performance liquid chromatography (HPLC) and thin-layer chromatography (TLC).