Heterologous expression and secretion of Lactobacillus amylovorus alpha-amylase in Leuconostoc citreum

Biotechnol Lett. 2009 Nov;31(11):1783-8. doi: 10.1007/s10529-009-0079-1. Epub 2009 Jul 18.

Abstract

To develop a gene expression system for Leuconostoc genus, construction of expression vector and expression of a heterologus protein in Leuconostoc was performed. Alpha-amylase gene from Lactobacillus amylovorus was cloned into a Leuconostoc cloning vector, pLeuCM, with its own signal peptide. pLeuCMamy was introduced into Leuconostoc citreum CB2567 and a successful expression of alpha-amy gene was confirmed by enzyme activity assays. About 90% of alpha-amylase activity was detected in the culture broth, revealing most of expressed alpha-amylase was secreted out cells. The signal sequence of alpha-amy gene is a good candidate for the secretion of heterologous protein by using Leuconostoc host-vector system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • Genes, Bacterial
  • Lactobacillus acidophilus / enzymology*
  • Lactobacillus acidophilus / genetics
  • Leuconostoc / cytology
  • Leuconostoc / growth & development
  • Leuconostoc / metabolism*
  • Plasmids / genetics
  • Restriction Mapping
  • Sequence Analysis, DNA
  • Transformation, Genetic
  • alpha-Amylases / genetics
  • alpha-Amylases / metabolism*

Substances

  • alpha-Amylases