Light‐induced cytochrome P450‐dependent enzyme in indole alkaloid biosynthesis: tabersonine 16‐hydroxylase

FEBS Letters

Unterbusch, Elke²  De Luca, Vincenzo³  Schmidt, Jürgen¹  Schröder, Joachim²  Strack, Dieter¹  Schröder, Gudrun²  Kaltenbach, Melanie² 

[1] Institut für Pflanzenbiochemie, Weinberg 3, D-06120 Halle (Saale), Germany;Institut für Biologie II, Universität Freiburg, Schänzlestrasse 1, D-79104 Freiburg, Germany;Novartis Agribusiness Biotechnology Research, Inc., P.O. Box 12257, Research Triangle Park, NC 27709-2257, USA

关键词: Alkaloid biosynthesis;    Cytochrome P450;    Indole alkaloid;    Light induction;    Tabersonine hydroxylase;    Catharanthus roseus;    T16H;    tabersonine 16-hydroxylase;    P450;    cytochrome P450 monooxygenase;    P450Red;    P450 reductase;    SAM;    S-adenosyl-L-methionine;    TLC;    thin layer chromatography;    D4H;    deacetoxyvindoline 4-hydroxylase;    DAT;    deacetylvindoline 4-O-acetyltransferase;    NMT;    N-methyltransferase;    OMT;    O-methyltransferase;   

DOI  :  10.1016/S0014-5793(99)01138-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Vinblastine and vincristine are two medically important bisindole alkaloids from Catharanthus roseus (Madagascar periwinkle). Attempts at production in cell cultures failed because a part of the complex pathway was not active, i.e. from tabersonine to vindoline. It starts with tabersonine 16-hydroxylase (T16H), a cytochrome P450-dependent enzyme. We now show that T16H is induced in the suspension culture by light and we report the cloning of the cDNA. The enzyme was expressed in Escherichia coli as translational fusion with the P450 reductase from C. roseus, and the reaction product was identified by mass spectrometry. The protein (CYP71D12) shares 47–52% identity with other members of the CYP71D subfamily with unknown function. The induction by light was strongly enhanced by a nutritional downshift (transfer into 8% aqueous sucrose). We discuss the possibility that the entire pathway to bisindoles can be expressed in suspension cultures.

【 授权许可】

Unknown