凝结芽孢杆菌AHU 1366膜中UDP-D-葡萄糖：N-乙酰葡糖胺焦磷酸十一碳烯醇葡糖基转移酶的增溶作用及性质

Solubilization and properties of UDP-D-glucose:N-acetylglucosaminyl pyrophosphorylundecaprenol glucosyltransferase from Bacillus coagulans AHU 1366 membranes.

作者信息

Kumita K, Murazumi N, Araki Y, Ito E

机构信息

Department of Chemistry, Faculty of Science, Hokkaido University.

出版信息

J Biochem. 1988 Dec;104(6):985-8. doi: 10.1093/oxfordjournals.jbchem.a122595.

DOI:10.1093/oxfordjournals.jbchem.a122595

PMID:2977388

Abstract

The glucosyltransferase which catalyzes the conversion of GlcNAc-PP-undecaprenol into Glc(beta 1----4)GlcNAc-PP-undecaprenol in the presence of UDP-glucose was solubilized from Bacillus coagulans AHU 1366 membranes by treatment with 0.1% Triton X-100 and partially purified by means of column chromatography on Sephacryl S-300 and DEAE-Sephacel. The final preparation was virtually free from other enzymes involved in the de novo synthesis of teichoic acid. The enzyme had a pH optimum of 6.6-8.0 and a Km value for UDP-glucose of 21 microM. The enzyme required 40 mM MgCl2, 0.6 M KCl, and 0.1% Nonidet P-40 for full activity.

摘要

在UDP-葡萄糖存在的情况下，催化GlcNAc-PP-十一异戊烯醇转化为Glc(β1----4)GlcNAc-PP-十一异戊烯醇的葡糖基转移酶，通过用0.1% Triton X-100处理从凝结芽孢杆菌AHU 1366膜中溶解出来，并通过Sephacryl S-300和DEAE-琼脂糖凝胶柱色谱法进行部分纯化。最终制剂几乎不含参与磷壁酸从头合成的其他酶。该酶的最适pH为6.6 - 8.0，UDP-葡萄糖的Km值为21μM。该酶需要40 mM MgCl2、0.6 M KCl和0.1% Nonidet P-40才能达到完全活性。