新型梭菌 O-乙酰高丝氨酸硫内酯酶的克隆、基因表达及酶学性质研究
O-acetylhomoserine sulfhydrylase from Clostridium novyi. Cloning, expression of the gene and characterization of the enzyme.
机构信息
Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, Vavilov Street, 32, Moscow, 119991, Russia.
Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, Vavilov Street, 32, Moscow, 119991, Russia.
出版信息
Protein Expr Purif. 2021 Apr;180:105810. doi: 10.1016/j.pep.2020.105810. Epub 2020 Dec 16.
The gene NT01CX_1210 of pathogenic bacterium Clostridium novyi annotated as encoding O-acetylhomoserine sulfhydrylase was cloned and expressed in Escherichia coli. The gene product having O-acetylhomoserine sulfhydrylase activity was purified to homogeneity. The protein showed molecular mass of approximately 184 kDa for the native form and 46 kDa for the subunit. The enzyme catalyzes the γ-substitution reaction of O-acetylhomoserine with maximum activity at pH 7.5. Analysis of C. novyi genome allowed us to suggest that there is only one way for the synthesis of l-methionine in the bacterium. The data obtained may provide the basis for further study of the role of OAHS in Clostridium bacteria and an ascertainment of its mechanism.
致病性细菌诺维梭菌的基因 NT01CX_1210 被注释为编码 O-乙酰高丝氨酸硫内酯酶，该基因在大肠杆菌中被克隆和表达。具有 O-乙酰高丝氨酸硫内酯酶活性的基因产物被纯化至均一状态。该蛋白的天然形式的分子量约为 184 kDa，亚基的分子量约为 46 kDa。该酶催化 O-乙酰高丝氨酸的γ取代反应，在 pH 7.5 时具有最大活性。对诺维梭菌基因组的分析使我们推测，在该细菌中只有一种合成 l-蛋氨酸的途径。获得的数据可能为进一步研究 OAHS 在梭菌中的作用及其机制提供依据。
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