Gene cloning, expression, and characterization of a novel trehalose synthase from Arthrobacter aurescens.
Trehalose synthase (TreS) is an intramolecular transglycosylase. It specially catalyzes the conversion of maltose and trehalose. In this study, a novel treS gene, which had a length of 1,797 bp and encoded 598 amino acids, was cloned from Arthrobacter aurescens CGMCC 1.1892 and expressed in Escherichia coli. Thin layer chromatography results indicated that it could catalyze the conversion between maltose and trehalose in one step. However, the ion chromatography results showed that, as a byproduct, about 13% glucose was also produced. The purified recombinant enzyme had a molecular weight of 68 kDa and showed its optimal activity at 35 degrees C and pH 6.5. This enzyme was not thermostable, and its activity was increased by 1 mM Mg2+, Mn2+, and Ca2+ while strongly inhibited by 5 mM Cu2+ and SDS.
Xiuli
Wu
W
Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
Hongbiao
Ding
D
Ming
Yue
Y
Yu
Qiao
Q
D001173:Arthrobacter / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005964:Glucosyltransferases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013379:Substrate Specificity
D001426:Bacterial Proteins; D005964:Glucosyltransferases; C099692:trehalose synthase