Skarlatos P, Dahl M K
Lehrstuhl für Mikrobiologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Germany.
J Bacteriol. 1998 Jun;180(12):3222-6. doi: 10.1128/JB.180.12.3222-3226.1998.
The open reading frame yqgR (now termed glcK), which had been sequenced as part of the genome project, encodes a glucose kinase of Bacillus subtilis. A 1.1-kb DNA fragment containing glcK complemented an Escherichia coli strain deficient in glucose kinase activity. Insertional mutagenesis of glcK resulted in a complete inactivation of glucose kinase activity in crude protein extracts, indicating that B. subtilis contains one major glucose kinase. The glcK gene encodes a 321-residue protein with a molecular mass of 33.5 kDa. The glucose kinase was overexpressed as a fusion protein to a six-His affinity tag and purified to homogeneity. The enzyme had K(m) values for ATP and glucose of 0.77 and 0.24 mM, respectively, and a Vmax of 93 mumol min-1 mg-1. A B. subtilis strain deficient for glucose kinase grew at the same rate on different carbon sources tested, including disaccharides such as maltose, trehalose, and sucrose.
作为基因组计划的一部分进行测序的开放阅读框yqgR（现称为glcK），编码枯草芽孢杆菌的一种葡萄糖激酶。一个包含glcK的1.1 kb DNA片段互补了一株葡萄糖激酶活性缺陷的大肠杆菌菌株。glcK的插入诱变导致粗蛋白提取物中的葡萄糖激酶活性完全失活，这表明枯草芽孢杆菌含有一种主要的葡萄糖激酶。glcK基因编码一个由321个残基组成、分子量为33.5 kDa的蛋白质。葡萄糖激酶作为与六聚组氨酸亲和标签的融合蛋白进行过表达，并纯化至同质。该酶对ATP和葡萄糖的K(m)值分别为0.77和0.24 mM，Vmax为93 μmol min-1 mg-1。一株缺乏葡萄糖激酶的枯草芽孢杆菌菌株在包括麦芽糖、海藻糖和蔗糖等二糖在内的不同测试碳源上以相同的速率生长。
