Covalent flavinylation of 6‐hydroxy‐D‐nicotine oxidase involves an energy‐requiring process

FEBS Letters

Bichler, Veronika¹  Brandsch, Roderich¹ 

[1] Biochemisches Institut der Universität, Hermann-Herder-Str. 7, D-7800 Freiburg, FRG

关键词: 6-Hydroxy-D-nicotine oxidase;    Covalent flavinylation;    Flavoenzyme;    FAD;    6-HDNO;    6-hydroxy-D-nicotine oxidase;    IPTG;    isopropyl-β-D-thiogalactopyranoside;    DPI;    diphenyleneiodonium;    HPLC;    high-performance liquid chromatography;    CCCP;    carbonylcyanide chlorophenylhydrazone;    PEP;    phosphoenolpyruvate;    PK;    pyruvate kinase;    Cm;    chloramphenicol;   

DOI  :  10.1016/0014-5793(87)80433-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

E. coli cells harbouring the recombinant plasmid pDB222 with the 6-HDNO gene under the control of the tac-promotor were induced with IPTG to synthesize a high amount of 6-HDNO protein. Part of this protein was present as 6-HDNO apoenzyme. The proportion of 6-HDNO apoenzyme formed could be increased when the induction of 6-HDNO synthesis by IPTG was performed in the presence of the inhibitor diphenyleneiodonium. The 6-HDNO apoenzyme thus formed could be transformed into enzymatically active holoenzyme in the presence of FAD by a process requiring an energy-generating system consisting of ATP, phosphoenolpyruvate and pyruvate kinase. This finding suggests that an enzymatic step(s) is (are) involved in the covalent flavinylation of 6-HDNO.

【 授权许可】

Unknown