KOPRI Repository

A new approach for discovering cold-active enzymes in a cell mixture of pure-cultured bacteria

Cited 5 time in scopus
Metadata Downloads
Title
A new approach for discovering cold-active enzymes in a cell mixture of pure-cultured bacteria
Authors
Kim, Dockyu
Park, Ha Ju
Kim, Il-Chan
Yim, Joung Han
Subject
Biotechnology & Applied Microbiology
Keywords
Activity-based enzyme screen; Chitinase; Cold-active; Culture mixture; Functional expression; Metagenomic library; Pseudoalteromonas issachenkonii
Issue Date
2014
Citation
Kim, Dockyu, et al. 2014. "A new approach for discovering cold-active enzymes in a cell mixture of pure-cultured bacteria". Biotechnology Letters, 36(3): 567-573.
Abstract
To overcome the intrinsic problems of conventional approaches, such as the unavailability of source microorganisms in metagenomic libraries and the production of inactive aggregates, a new method was tested for discovering new enzymes (e.g. coldactive chitinase). A metagenome-like library was constructed using genomes extracted from a cell mixture of pure-cultured chitinolytic bacteria, followed by activity-based screening for Escherichia coli clones that exhibit chitinase activity on selective medium. Within one positive chitinolytic clone, one chitinase gene (chi22718_III) was detected and assigned to the arctic marine bacterium, Pseudoalteromonas issachenkonii PAMC 22718, by colony-PCR with chi22718_III-specific primers. When expressed in E. coli, recombinant R-Chi22718_III lost 85 % of its enzyme activity when pre-incubated at 40 ℃ for 1 h, whereas its mesophilic counterpart R-ChiK only lost 10 % of its activity under the same conditions indicating that R-Chi22718_III is thermolabile, a characteristic of cold-active enzymes.
DOI
http://dx.doi.org/10.1007/s10529-013-1384-2
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