Effects of replacement of promoters and modification of the leader peptide region of the amy gene of Streptomyces griseus on synthesis and secretion of alpha-amylase by Streptomyces lividans

Mol Gen Genet. 1991 Dec;231(1):88-96. doi: 10.1007/BF00293826.

Abstract

Five different mutations were introduced into the leader peptide region of the alpha-amylase gene of Streptomyces griseus IMRU 3570. A mutation which increased the positive charge of the N-terminal region of the leader peptide enhanced the secretion of alpha-amylase by two- to threefold. Replacement of the native promoter of the amylase gene by the promoter of the Tn5 neo gene or by the promoter of the saf gene resulted in a 16-fold increase in alpha-amylase secretion. The enhanced secretion of alpha-amylase obtained by using the most efficient promoters was due to a correlated increase in the amount of transcript formed. The translation and secretion processes in S. lividans are not a bottleneck for enzyme secretion even at very high transcription rates, since stimulation of transcription of the alpha-amylase gene results in a proportionate increase in secretion of the enzyme.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Gene Expression / genetics
  • Kinetics
  • Molecular Sequence Data
  • Mutagenesis / genetics
  • Mutation / genetics
  • Plasmids / genetics
  • Promoter Regions, Genetic / genetics*
  • Protein Sorting Signals / chemistry
  • Protein Sorting Signals / genetics*
  • Streptomyces / enzymology
  • Streptomyces / genetics*
  • Streptomyces griseus / enzymology
  • Streptomyces griseus / genetics
  • alpha-Amylases / biosynthesis
  • alpha-Amylases / genetics*
  • alpha-Amylases / metabolism

Substances

  • Protein Sorting Signals
  • alpha-Amylases