Purification of Clostridium thermocellum xylanase Z expressed in Escherichia coli and identification of the corresponding product in the culture medium of C. thermocellum - PubMed (original) (raw)

Purification of Clostridium thermocellum xylanase Z expressed in Escherichia coli and identification of the corresponding product in the culture medium of C. thermocellum

O Grépinet et al. J Bacteriol. 1988 Oct.

Abstract

An endoxylanase encoded by the xynZ gene of Clostridium thermocellum was purified from Escherichia coli harbouring a fragment of the gene cloned in pUC8. The purified enzyme showed two active bands of Mr 41,000 and 39,000, the latter one presumably derived from the former through proteolysis. The enzyme was highly active on xylan and para-nitrophenyl-beta-D-xylobioside. The major end product of xylan hydrolysis was xylobiose. With an antiserum raised against the enzyme purified from E. coli, an immunoreactive polypeptide of Mr 90,000, corresponding to the entire xynZ gene product, was detected in a culture supernatant from C. thermocellum grown on cellulose. By immunological detection, xylanase Z was shown to be associated with a cellulose-binding, high-molecular-weight fraction whose properties coincided with those described previously for the cellulose-degrading complex of C. thermocellum known as the cellulosome.

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References

1. Anal Biochem. 1983 Jun;131(2):333-6 - PubMed
1. Nature. 1970 Aug 15;227(5259):680-5 - PubMed
1. Cell. 1985 Jan;40(1):9-17 - PubMed
1. Appl Environ Microbiol. 1985 Mar;49(3):656-9 - PubMed
1. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 - PubMed

Purification of Clostridium thermocellum xylanase Z expressed in Escherichia coli and identification of the corresponding product in the culture medium of C. thermocellum - PubMed (original) (raw)