Secretion of the recombination α-amylase in Escherichia coli and purification by the gram-positive enhancer matrix (GEM) particles

Abstract

α-Amylases are important enzymes in industry. A recombinant α-amylase with a secretion signal peptide and an AcmA tag was expressed in Escherichia coli to improve the yield. The induction concentrations were optimized, and the temperature had a significant influence on soluble expression and secretion. A visible band could be obtained when the induction was conducted at 16 °C. The gram-positive enhancer matrix (GEM) particles could separate and purify the recombinant α-amylase with the AcmA tag, and no visible band could be seen in the culture even after the culture was concentrated ten times. The solution and concentration of the recombinant α-amylase could be adjusted by GEM particles. The recombinant untagged α-amylase was obtained after digestion. The α-amylase was characterized. The recombinant α-amylase was a thermophilic enzyme with a broad pH tolerance. In addition, the enzyme activity of the recombinant α-amylase was independent of Ca2+. The recombinant α-amylase contained the OmpA signal peptide and the AcmA tag and was expressed and purified quickly and easily.