Molecular Cloning and Characterization of a Novel α-Amylase from Antarctic Sea Ice Bacterium Pseudoalteromonas sp. M175 and Its Primary Application in Detergent. (27th June 2018)

• Record Type: Journal Article
• Authors: Wang, Xiaofei; Kan, Guangfeng; Ren, Xiulian; Yu, Geng; Shi, Cuijuan; Xie, Qiuju; Wen, Hua; Betenbaugh, Michael
• Other Names: Shi Pengjun Academic Editor.
• Is Part Of: BioMed research international. Volume 2018(2018)
• Publication Date: 2018-06-27
• Subjects: Medicine -- Periodicals; Biology -- Periodicals; Biotechnology -- Periodicals; Life sciences -- Periodicals; 610.5
• DOI: 10.1155/2018/3258383
• Languages: English
• ISSNs: 2314-6133

Abstract

Abstract : A novel cold-adapted and salt-tolerant α -amylase gene ( amy 175) from Antarctic sea ice bacterium Pseudoalteromonas sp. M175 was successfully cloned and expressed. The open reading frame (ORF) of amy 175 had 1722 bp encoding a protein of 573 amino acids residues. Multiple alignments indicated Amy175 had seven highly conserved sequences and the putative catalytic triad (Asp 244, Glu 286, and Asp 372 ). It was the first identified member of GH13_36 subfamily which contained QPDLN in the CSR V. The recombinant enzyme (Amy175) was purified to homogeneity with a molecular mass of about 62 kDa on SDS-PAGE. It had a mixed enzyme specificity of α -amylase and α -glucosidase. Amy175 displayed highest activity at pH 8.0 and 25°C and exhibited extreme salt-resistance with the maximum activity at 1 M NaCl. Amy175 was strongly stimulated by Mg 2+, Ni 2+, K +, 1 mM Ca 2+, 1 mM Ba 2+, 1 mM Pb 2+, 1 mM sodium dodecyl sulphate (SDS), and 10% dimethyl sulfoxide (DMSO) but was significantly inhibited by Cu 2+, Mn 2+, Hg 2+, 10 mM β -mercaptoethanol ( β -ME), and 10% Tween 80. Amy175 demonstrated excellent resistance towards all the tested commercial detergents, and wash performance analysis displayed that the addition of Amy175 improved the stain removal efficiency. This study demonstrated that Amy175 would be proposed as a novel α -amylase source for industrial application in the future.