In vivo and in vitro expression of the 6‐hydroxy‐D‐nicotine oxidase gene of Arthrobacter oxidans, cloned into Escherichia coli, as an enzymatically active, covalently flavinylated polypeptide

FEBS Letters

Bichler, Veronika¹  Brandsch, Roderich¹ 

[1] Biochemisches Institut der Universität, D-7800 Freiburg i.Br., FRG

关键词: (Arthrobacter oxidans);    6-Hydroxy-D-nicotine oxidase;    Covalent flavinylation;    Coupled transcription-translation;    6-HDNO;    6-hydroxy-D-nicotine oxidase;    SDH;    succinate dehydrogenase;    FR;    fumarate reductase;    DNBD;    dinucleotide-binding domain;    kb;    kilobases;   

DOI  :  10.1016/0014-5793(85)80108-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The 6-hydroxy-D-nicotine oxidase gene of Arthrobacter oxidans was cloned into E.coli with the aid of the expression vector pKK-223-3. This enzyme, as well as the E.coli enzymes succinate dehydrogenase and fumarate reductase, bears the cofactor FAD covalently attached to the polypeptide through a His-N₃-8α-linkage. The amino acid sequence surrounding the histidine residue involved in FAD binding in 6-hydroxy-D-nicotine oxidase and the two E.coli enzymes, however, show no homology. Nevertheless, 6-hydroxy-D-nicotine oxidase is expressed in E.coli in vivo and in an E.coli-derived coupled transcription-translation system as a covalently flavinylated, enzymatically active polypeptide.

【 授权许可】

Unknown