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Identification of three distinct Clostridium thermocellum xylanase genes by molecular cloning

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1007/BF00425176
AuthorSearch for: 1; Search for: 1; Search for: 1; Search for: 1; Search for: 1
Affiliation
  1. National Research Council Canada. NRC Institute for Biological Sciences
FormatText, Article
Abstract
Three genes coding for xylanase synthesis in Clostridium thermocellum were cloned and expressed in Escherichia coli. Genomic DNA from Clostridium thermocellum was digested to completion with HindIII, BamHI, and SalI. The fragments were ligated into the corresponding sites of pUC19 and transformed into Escherichia coli. Two of the genes encoded for xylanases which depolymerized xylans but were unable to extensively convert these substrates to reducing sugar. The third gene encoded for an enzyme that extensively hydrolyzed xylan. The insert containing the latter gene was subjected to extensive mapping and was found to encode for a xylanase with a molecular weight of approximately 25,000. The protein product of the cloned gene was obtained in a relatively pure form by heat treatment, ion exchange and gel permeation steps. The enzyme was quite stable to high temperatures with a half-life of 24 h at 70°C.
Publication date
In
  • Archives of Microbiology 152, no. 4 (1989): 377–381.
LanguageEnglish
NRC numberMACKENZIE1989
NRCC 30545
NPARC number9381259
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Record identifiera8f878de-c2bf-4b70-ab5d-c7c5468acb5c
Record created2009-07-10
Record modified2020-03-17
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