Interdisciplinary Graduate School of Science and Technology, Department of Life Sciences, Shinshu University, 3-15-1 Tokida, Ueda, Nagano 386-8567, Japan.
Biochem Biophys Res Commun. 2010 Jul 30;398(3):606-12. doi: 10.1016/j.bbrc.2010.07.001. Epub 2010 Jul 4.
CwlQ (previous YjbJ) is one of the putative cell wall hydrolases in Bacillus subtilis. Its domain has an amino acid sequence similar to the soluble-lytic transglycosylase (SLT) of Escherichia coli Slt70 and also goose lysozyme (muramidase). To characterize the enzyme, the domain of CwlQ was cloned and expressed in E. coli. The purified CwlQ protein exhibited cell wall hydrolytic activity. Surprisingly, RP-HPLC, mass spectrometry (MS), and MS/MS analyses showed that CwlQ produces two products, 1,6-anhydro-N-acetylmuramic acid and N-acetylmuramic acid, thus indicating that CwlQ is a bifunctional enzyme. The site-directed mutagenesis revealed that glutamic acid 85 (Glu-85) is an amino acid residue essential to both activities.
CwlQ(以前称为 YjbJ)是枯草芽孢杆菌中假定的细胞壁水解酶之一。它的结构域具有与大肠杆菌 Slt70 的可溶性溶菌酶(SLT)和鹅溶菌酶(胞壁质酶)相似的氨基酸序列。为了表征该酶,在大肠杆菌中克隆并表达了 CwlQ 的结构域。纯化的 CwlQ 蛋白表现出细胞壁水解活性。令人惊讶的是,反相高效液相色谱法(RP-HPLC)、质谱(MS)和 MS/MS 分析表明,CwlQ 产生两种产物,1,6-脱水-N-乙酰胞壁酸和 N-乙酰胞壁酸,因此表明 CwlQ 是一种双功能酶。定点突变揭示谷氨酸 85(Glu-85)是两个活性都必需的氨基酸残基。