Purification and characterization of methylmalonyl-CoA mutase from a methanol-utilizing bacterium, Methylobacterium extorquens NR-1.
Purification and characterization of methylmalonyl-CoA mutase from a methanol-utilizing bacterium, Methylobacterium extorquens NR-1.
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来自利用甲醇的细菌扭扭甲基杆菌 NR-1 的甲基丙二酰辅酶 A 变位酶的纯化和表征。
DOI：
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发表时间：
2002
影响因子：
1.6
通讯作者：
Y. Nakano
中科院分区：
文献类型：
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作者：
E. Miyamoto;F. Watanabe;R. Yamaji;H. Inui;Kazuyoshi Sato;Y. Nakano
在利用甲醇的细菌扭脱甲基杆菌NR-1的细胞生长期间，不断发现甲基丙二酰-CoA酶（82-95%脱辅基酶）的高活性（约50 mU × mg蛋白（-1））。将脱辅基酶纯化至均一并表征。纯化的酶是无色的。一个明显的M。通过Superdex 200 HR凝胶过滤，计算出extroquens NR-1酶为150，000 +/-5，000。纯化酶的SDS-聚丙烯酰胺凝胶电泳显示两条蛋白带，其表观Mr分别为85.000 ± 2，000和70，000 ± 2，000，表明M.扭脱体NR-1酶由两个不同的亚基组成。M.的小亚基和大亚基的NH 2-末端氨基酸序列。extroquens NR-1酶与其他物种的NR-1酶没有明显的同源性。对该菌的酶学性质进行了研究。研究了扭脱醌NR-1酶的活性。
High activity (about 50 mU x mg protein(-1)) of methylmalonyl-CoA mutase (82-95% apo-enzyme) was constantly found during the cell growth of a methanol-utilizing bacterium, Methylobacterium extorquens NR-1. The apo-enzyme was purified to homogeneity and characterized. The purified enzyme was colorless. An apparent Mr of M. extorquens NR-1 enzyme was calculated to be 150,000 +/- 5,000 by Superdex 200 HR gel filtration. SDS-polyacrylamide gel electrophoresis of the purified enzyme gave two protein bands with an apparent Mr of 85.000 +/- 2,000 and 70,000 +/- 2,000, indicating that the M. extorquens NR-1 enzyme is composed of two nonidentical subunits. NH2-terminal amino acid sequences of the small and large subunits of M. extorquens NR-1 enzyme showed no significant homology to those of the enzyme from other species. Some enzymological properties of the M. extorquens NR-1 enzyme were studied.