In vitro expression and activity of lycopene cyclase and β‐carotene hydroxylase from Erwinia herbicola

FEBS Letters

Beyer, Peter¹  O'Brien, David A.²  Kiemig, Hans¹  Hearst, John E.²  Hundle, Bhupinder S.² 

[1] Institut für Biologie 2, Albert-Ludwigs-Universität, Schänzlestr. 1, D-7800 Freiburg, Germany;Department of Chemistry, University of California and Division of Chemical Biodynamics, Lawrence Berkeley Laboratory, Berkeley, CA 94720, USA

关键词: Carotenoid;    Zeaxanthin;    Lycopene;    β-Carotene;    Erwinia herbicola;    Cyclase;    Hydroxylase;    Overexpression;    HPLC;    high performance liquid chromatography;    PCR;    polymerase chain reaction;    SDS-PAGE;    sodium dodecylsulfate polyacrylamide gel electrophoresis;    IPTG;    isopropyl β-D-thio-galac-topyranoside;    βME;    β-mercaptoethanol;    PMSF;    phenylmethylsulfonyl fluoride;   

DOI  :  10.1016/0014-5793(93)81188-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The cyclisation of lycopene to β-carotene and the hydroxylation of β-carotene to zeaxanthin are common enzymatic steps in the biosynthesis of carotenoids in a wide range of bacteria, fungi, and plants. We have individually expressed in E. coli the two genes coding for these enzymatic steps in Erwinia herbicola. The cyclase and hydroxylase enzymes have apparent molecular weights of 43 kDa and 22 kDa, respectively, as determined by SDS-PAGE. Hydroxylase in vitro activity was obtained only in the cytoplasmic fraction. Cyclase also demonstrated enzyme activity in a crude cell-free lysate, although to a lesser extent.

【 授权许可】

Unknown