title: Characterization of an organic solvent-tolerant α-amylase from a halophilic isolate, Thalassobacillus sp. LY18.
abstract: A halophilic isolate Thalassobacillus sp. LY18 producing extracellular amylase was isolated from the saline soil of Yuncheng Salt Lake, China. Production of the enzyme was synchronized with bacterial growth and reached a maximum level during the early stationary phase. The amylase was purified to homogeneity with a molecular mass of 31 kDa. Major products of soluble starch hydrolysis were maltose and maltotriose, indicating an α-amylase activity. Optimal enzyme activity was found to be at 70°C, pH 9.0, and 10 % NaCl. The α-amylase was highly stable over broad temperature (30-90°C), pH (6.0-12.0), and NaCl concentration (0-20 %) ranges, showing excellent thermostable, alkalistable, and halotolerant nature. The enzyme was stimulated by Ca(2+), but greatly inhibited by EDTA, indicating it was a metalloenzyme. Complete inhibition by diethyl pyrocarbonate and β-mercaptoethanol revealed that histidine residue and disulfide bond were essential for enzyme catalysis. The surfactants tested had no significant effects on the amylase activity. Furthermore, it showed high activity and stability in the presence of water-insoluble organic solvents with log P (ow) ≥ 2.13.
authors.affiliation: Life Science College, Yuncheng University, Yuncheng 044000, China. lixin-eva@163.com
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.forename: Hui-Ying
authors.initials: HY
authors.lastname: Yu
mesh_terms: D001406:Bacillaceae / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002681:China; D004795:Enzyme Stability; D006358:Hot Temperature; D060106:Lakes / Q000382:microbiology*; D013213:Starch / Q000378:metabolism; D000516:alpha-Amylases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D013213:Starch; D000516:alpha-Amylases