Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, College of Life Science, Nanjing Agricultural University, Nanjing, 210095, PR China.
College of Agriculture, Xinjiang Agricultural University, XinJiang, 830052, China.
Protein Expr Purif. 2021 Jun;182:105846. doi: 10.1016/j.pep.2021.105846. Epub 2021 Feb 13.
Trehalase catalyzes the hydrolysis of trehalose into two glucose molecules and is present in nearly all tissues in various forms. In this study, a putative bacterial trehalase gene, encoding a glycoside hydrolase family 15 (GH15) protein was identified in Microvirga sp. strain MC18 and heterologously expressed in E. coli. The specific activity of the purified recombinant trehalase MtreH was 24 U/mg, with K and V values of 23.45 mg/mL and 184.23 μmol/mg/min, respectively. The enzyme exhibited optimal activity at 40 °C and pH 7.0, whereby Ca had a considerable positive effects on the catalytic activity and thermostability. The optimized enzymatic reaction conditions for the bioconversion of trehalose using rMtreH were determined as 40 °C, pH 7.0, 10 h and 1% trehalose concentration. The characterization of this bacterial trehalase improves our understanding of the metabolism and biological role of trehalose in prokaryotic organism.
海藻糖酶催化海藻糖水解为两个葡萄糖分子，存在于各种形式的几乎所有组织中。在这项研究中，从 Microvirga 菌株 MC18 中鉴定出一种假定的细菌海藻糖酶基因，该基因编码糖苷水解酶家族 15（GH15）蛋白，并在大肠杆菌中异源表达。纯化的重组海藻糖酶 MtreH 的比活性为 24 U/mg，K 和 V 值分别为 23.45 mg/mL 和 184.23 μmol/mg/min。该酶在 40°C 和 pH 7.0 下表现出最佳活性，其中 Ca 对催化活性和热稳定性有显著的积极影响。使用 rMtreH 进行海藻糖生物转化的最佳酶反应条件确定为 40°C、pH 7.0、10 h 和 1%海藻糖浓度。这种细菌海藻糖酶的特性提高了我们对海藻糖在原核生物中的代谢和生物学作用的理解。
