节杆菌属中一种果糖基胺氧化酶的分离与鉴定

Isolation and characterization of a fructosyl-amine oxidase from an Arthrobacter sp.

作者信息

Ferri Stefano, Sakaguchi Akane, Goto Hiroki, Tsugawa Wakako, Sode Koji

机构信息

Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, 184-8588, Tokyo, Koganei, Japan.

出版信息

Biotechnol Lett. 2005 Jan;27(1):27-32. doi: 10.1007/s10529-004-6312-z.

DOI:10.1007/s10529-004-6312-z

PMID:15685416

Abstract

An Arthrobacter sp. was isolated that, when induced by fructosyl-valine, expressed a fructosyl-amine oxidase (FAOD) that was specific for alpha-glycated amino acids. The N-terminal amino acid sequence of the purified oxidase was determined and used to design oligonucleotides to amplify the gene by inverse PCR. Expression of the gene in Escherichia coli produced 0.23 units FAOD per mg protein, over 30-fold greater than native expression levels, with properties almost indistinguishable from the native enzyme. The presence of FAOD was confirmed in other Arthrobacter ssp.

摘要

分离出了一种节杆菌属细菌，当用果糖基缬氨酸诱导时，它会表达一种对α-糖化氨基酸具有特异性的果糖基胺氧化酶（FAOD）。测定了纯化氧化酶的N端氨基酸序列，并用于设计寡核苷酸通过反向PCR扩增该基因。该基因在大肠杆菌中的表达产生了每毫克蛋白质0.23单位的FAOD，比天然表达水平高30多倍，其性质与天然酶几乎无法区分。在其他节杆菌属细菌中也证实了FAOD的存在。