Sequence structure and expression of a cloned β-glucosidase gene from an extreme thermophile

D. R. Love, R. Fisher, P. L. Bergquist*

*Corresponding author for this work

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

The gene for a β-glucosidase from the extremely thermophilic bacterium Caldocellum saccharolyticum has been isolated from a genomic library and sequenced. An open reading frame identified by computer analysis of the sequence could encode a protein of Mr 54400, which is close to the size of the polypeptide experimentally determined using maxicells. Analysis of the amino-terminal residues of the protein produced in Escherichia coli suggests that it is processed by a methionine aminopeptidase. A sequence within C. saccharolyticum DNA upstream of the β-glucosidase gene was found to act as a promoter for expression of the thermophile gene in E. coli. The protein has been overproduced in E. coli and Bacillus subtilis where it retains its enzymatic activity and heat stability. There appears to be a single copy of the gene in Caldocellum DNA.

Original languageEnglish
Pages (from-to)84-92
Number of pages9
JournalMGG Molecular & General Genetics
Volume213
Issue number1
DOIs
Publication statusPublished - Jul 1988
Externally publishedYes

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Keywords

  • β-Glucosidase
  • Expression vectors
  • Sequence analysis
  • Thermophile

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