Cloning and expression of the α-amylase gene from Bacillus stearothermophilus in several staphylococcal species

Karin Thudt, Karl Heinz Schleifer, Friedrich Götz

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12 Scopus citations

Abstract

The plasmid-coded α-amylase gene of Bacillus stearothermophilus (amy) was cloned in Staphylococcus camosus using plasmid pCA43 as a vector. The amy gene was located on a 5.4-kb HindIII DNA fragment of the hybrid plasmid pamy7. When transformed into other staphylococcal species, plasmid pamy7 exhibited marked differences in the production of α-amylase (αAmy). Most active for heterospecific αAmy production was Staphylococcus aureus. In its culture supernatant nearly half as much αAmy activity was found as for the donor strain B. stearothermophilus. All staphylococcal species were able to secrete αAmy, since more than 80% of the enzyme activity was found in the culture supernatant. The extracellular αAmy of S. aureus[pamy7] was purified to homogeneity. The enzyme exhibited an Mr of approx. 58000, an optimum activity at pH 5.3-6.3 and at 65 °C. Although the enzyme was stable at 65 °C for at least 3 h, its thermostability was not unusual. The enzymatic properties of the αAmy from S. aureus were similar to those previously reported for various B. stearothermophilus strains.

Original languageEnglish
Pages (from-to)163-169
Number of pages7
JournalGene
Volume37
Issue number1-3
DOIs
StatePublished - 1985
Externally publishedYes

Keywords

  • Recombinant DNA
  • S. carnosus
  • Staphylococcus aureus
  • enzyme stability
  • plasmid vector pCA43
  • protein secretion
  • purification procedure

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