High-level expression, purification and characterization of a recombinant medium-temperature alpha-amylase from Bacillus subtilis

Biotechnol Lett. 2010 Jan;32(1):119-24. doi: 10.1007/s10529-009-0112-4. Epub 2009 Sep 1.

Abstract

Alpha-amylases are important industrial enzymes with a wide range of applications. Although medium-temperature alpha amylase (AmyE) has some practical advantages, its low yield has limited its applications. When an amyE gene from Bacillus subtilis BF768 was cloned into vector pWB980 and over-expressed in B. subtilis WB600, high activities (723 U ml(-1)) of secreted AmyE were produced. Recombinant AmyE was purified to a specific activity of 36 U mg(-1) having optimal activity at pH 6.0 and 60 degrees C.

MeSH terms

  • Bacillus subtilis / enzymology*
  • Bacillus subtilis / genetics
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Blotting, Western
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Temperature
  • alpha-Amylases / genetics
  • alpha-Amylases / metabolism*

Substances

  • Bacterial Proteins
  • alpha-Amylases