Cloning, expression, and carbon catabolite repression of the bamM gene encoding beta-amylase of Bacillus megaterium DSM319

Appl Microbiol Biotechnol. 2001 Jul;56(1-2):205-11. doi: 10.1007/s002530100645.

Abstract

The bamM gene from Bacillus megaterium DSM319 encoding an extracellular beta-amylase was isolated and completely sequenced. Chromosomal inactivation by deletion mutagenesis resulted in total loss of amylolytic activity, indicative of a single starch-degrading enzyme. Functional characterization of the expressed protein revealed a maltogenic enzyme exhibiting optimal activities at pH 7.5 and 50 degrees C. Amylase expression is subject to catabolite repression by glucose. A putative cis-acting catabolite-responsive element (CRE) was identified; it is located within the bamM coding region, matching the position of the predicted signal peptide processing site. Base substitutions introduced by site-directed mutagenesis within the bamM-CRE--retaining unchanged the amino acid sequence--provoked a remarkable relief from carbon catabolite repression (CCR), thereby proving functionality of the CRE for CCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacillus megaterium / enzymology*
  • Bacillus megaterium / genetics
  • Base Sequence
  • Carbon / metabolism*
  • Cloning, Molecular
  • Enzyme Repression*
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Response Elements*
  • beta-Amylase / genetics*

Substances

  • Carbon
  • beta-Amylase