Identification of α-amylase by random and specific mutagenesis of Texcoconibacillus texcoconensis 13CCT strain isolated from extreme alkaline-saline soil of the former Lake Texcoco (Mexico)

Folia Microbiol (Praha). 2014 May;59(3):235-40. doi: 10.1007/s12223-013-0289-8. Epub 2013 Nov 3.

Abstract

The alkaline α-amylase produced by Texcoconibacillus texcoconensis 13CC(T) strain was identified by random mutagenesis and confirmed by directed mutagenesis. A transposon mutagenesis approach was taken to identify the gene responsible for the degradation of starch in T. texcoconensis 13CC(T) strain. The deduced amino acids of the amy gene had a 99% similarity with those of Bacillus selenitireducens MLS10 and 97% with those of Paenibacillus curdlanolyticus YK9. The enzyme showed a maximum activity of 131.1 U/mL at 37 °C and pH 9.5 to 10.5. In situ activity of the enzyme determined by polyacrylamide gel electrophoresis showed only one band with amylolytic activity. This is the first report of a bacterium isolated from the extreme alkaline-saline soil of the former Lake Texcoco (Mexico) with amylolytic activity in alkaline conditions while its potential as a source of amylases for the industry is discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus / enzymology*
  • Bacillus / genetics*
  • Bacillus / growth & development
  • Bacillus / isolation & purification
  • Lakes
  • Mexico
  • Mutagenesis*
  • Mutation
  • Salinity*
  • Soil / chemistry*
  • Soil Microbiology*
  • alpha-Amylases / genetics
  • alpha-Amylases / metabolism*

Substances

  • Soil
  • alpha-Amylases