来自奥氏无色杆菌的α-氨基-ε-己内酰胺消旋酶的纯化及性质

Purification and properties of alpha-amino-epsilon-caprolactam racemase from Achromobacter obae.

作者信息

Ahmed S A, Esaki N, Soda K

出版信息

FEBS Lett. 1982 Dec 27;150(2):370-4. doi: 10.1016/0014-5793(82)80770-9.

Abstract

We have purified a unique enzyme, alpha-amino-epsilon-caprolactam racemase 945-fold from an extract of Achromobacter obae by Octyl-Sepharose CL-4B and Thiopropyl-Sepharose 6B and some other chromatographies. The purified enzyme was found homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and analytical ultracentrifugation. The enzyme has a monomeric structure with Mr approximately 50000 and a sedimentation coefficient (S20,w) of 4.28 S. The enzyme contains pyridoxal 5'-phosphate as a coenzyme. The pH optimum for the enzyme activity is approximately 9.0. D- and L-alpha-amino-epsilon-caprolactams are the only substrates. The Km values for the D- and L-isomers are, 8 and 6 mM, respectively.

摘要

我们通过辛基琼脂糖凝胶CL-4B、硫丙基琼脂糖凝胶6B及其他一些色谱法，从奥氏无色杆菌提取物中纯化出一种独特的酶，α-氨基-ε-己内酰胺消旋酶，纯化倍数达945倍。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和分析超速离心发现纯化后的酶呈均一状态。该酶具有单体结构，分子量约为50000，沉降系数（S20,w）为4.28 S。该酶含有磷酸吡哆醛作为辅酶。酶活性的最适pH约为9.0。D-和L-α-氨基-ε-己内酰胺是仅有的底物。D-和L-异构体的米氏常数分别为8 mM和6 mM。