KOPRI Repository

Expression of recombinant endochitinase from the Antarctic bacterium, Sanguibacter antarcticus KOPRI 21702 in Pichia pastoris by codon optimization

Cited 20 time in scopus
Metadata Downloads
Title
Expression of recombinant endochitinase from the Antarctic bacterium, Sanguibacter antarcticus KOPRI 21702 in Pichia pastoris by codon optimization
Other Titles
코돈최적화에 의한 남극세균 상귀박터 안탁티쿠스 코프리 21702 유래의 내재키티네이즈의 피키아 파스토리스 내에서의 재조합 발현
Authors
Koh, Hye Yeon
Kim, Il-Chan
Lee, Sung Gu
Na, Deuk Chae
Yim, Joung Han
Lee, Hong Kum
Park, Heeyong
Han, Se Jong
Subject
Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology
Keywords
Antarctica; Codon optimization; Endochitinases; Pichia pastoris; Recombinant
Issue Date
2010
Publisher
Elsevier
Citation
Koh, Hye Yeon, et al. 2010. "Expression of recombinant endochitinase from the Antarctic bacterium, Sanguibacter antarcticus KOPRI 21702 in Pichia pastoris by codon optimization". PROTEIN EXPRESSION AND PURIFICATION, 71(1): 108-114.
Abstract
An endochitinase was previously purified and the gene was cloned from the psychrophilic Antarctic bacterium,Sanguibacter antarcticus (KCTC 13143). In the present study, recombinant endochitinase,rChi21702, was expressed using a yeast expression system (Pichia pastoris) and codon optimization. The expressed rChi21702 was purified by Phenyl-Sepharose column chromatography. Optimal expression yielded 1-mg purified enzyme from 1-L bioreactor culture. When p-NP-(GlcNAc)2 was used as a substrate, the specific activity of the enzyme was determined to be 20 U/mg. In vitro assays and thin-layer chromatography demonstrated that the recombinant enzyme has endochitinase activity that produces diacetyl-chitobiose as a dominant end product when chitooligomers, colloidal chitin, and the chromogenic p-NP-(GlcNAc)2 are used as substrates. Optimal activity for rChi21702 was observed at 37oC and a pH of 7.6. Interestingly, rChi21702 exhibited 63% of optimal activity at 10oC and 44% activity at 0oC. Taken together, the results indicate that rChi21702 has psychrotolerant endochitinase activity even after recombinant expression in yeast cells.
URI
http://repository.kopri.re.kr/handle/201206/6294
DOI
http://dx.doi.org/10.1016/j.pep.2010.01.017
Files in This Item
General Conditions
      ROMEO Green
    Can archive pre-print and post-print or publisher's version/PDF
      ROMEO Blue
    Can archive post-print (ie final draft post-refereeing) or publisher's version/PDF
      ROMEO Yellow
    Can archive pre-print (ie pre-refereeing)
      ROMEO White
    Archiving not formally supported

    qrcode

    Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

    Browse