An extremely thermostable maltogenic amylase from Staphylothermus marinus: Bacillus expression of the gene and its application in genistin glycosylation

Abstract

The most extremely thermostable maltogenic amylase (SMMA) from archaeon Staphylothermus marinus has many potential applications in food processing. To ensure safety of microbial origin, a recombinant plasmid containing the enzymic gene and a constitutive promoter AmyR2 was constructed, and then transformed into a GRAS microorganism Bacillus subtilis. The purified SMMA from the liquid cultures of Bacillus has a specific activity of 66.96 U/mg, two times more than that from Escherichia coli SMMA was further employed to catalyze the genistion glycosylation using gamma-CD as both glucosyl donors and solubilizer. Glycosylated genistins with one to four additional alpha-glucosyls and a molar percentage of 69.87% in genistin reaction mixture were identified and quantified by HPLC-UV-MS. The glycosylated genistins at 0.2-1.2 mM showed an enhanced DPPH free radical scavenging capacity. To our knowledge, this is the first report on the Bacillus expression of archaeal maltogenic amylase. (C) 2017 Elsevier B.V. All rights reserved