Sequence of the gene encoding a highly thermostable neutral proteinase from Bacillus sp. strain EA1: expression in Escherichia coli and characterisation

David J. Saul*, Liam C. Williams, Helen S. Toogood, Roy M. Daniel, Peter L. Bergquist

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

The gene for a highly thermostable neutral proteinase (Npr) was isolated from Bacillus sp, strain EA1 by the polymerase chain reaction using consensus primers based on the sequences of npr genes from related species. The gene was sequenced and shown to be closely related to a neutral proteinase gene from Bacillus caldolyticus strain YP-T: the mature form of the enzyme differing by only a single amino acid. Enzyme samples were prepared from both the native organisms and also from recombinant Escherichia coli expressing the two npr genes. The proteinase from strain EA1 was shown to be significantly more thermostable than that from B. caldolyticus and that this difference is the result of a single amino acid substitution which is situated proximal to a region of the enzyme known to be crucial to conferring thermal stability. The phylogenetic relationship of EA1 to other Bacilli is also described.

Original languageEnglish
Pages (from-to)74-80
Number of pages7
JournalBiochimica et Biophysica Acta - Gene Structure and Expression
Volume1308
Issue number1
DOIs
Publication statusPublished - 31 Jul 1996

Keywords

  • (Bacillus)
  • Neutral proteinase
  • Thermophile

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