Heterologous expression of the plant proteins strictosidine synthase and berberine bridge enzyme in insect cell culture.
The heterologous expression of cDNAs encoding the alkaloid biosynthetic enzymes, strictosidine synthase [EC 4.3.3.2] from Rauvolfia serpentina and the berberine bridge enzyme [(S)-reticuline: oxygen oxidoreductase (methylene-bridge-forming), EC 1.5.3.9] from Eschscholtzia californica, has been achieved in a cell culture (Sf9) of the fall army worm, Spodoptera frugiperda, using a baculovirus-based expression system. The expression resulted in the overproduction of each plant enzyme in a catalytically active form. The maximal production attained was 4 mg purified, active enzyme per litre cell culture for both the strictosidine synthase and berberine bridge enzymes.
Kutchan
T M
TM
Laboratorium für Molekulare Biologie, Universität München, Germany.
Bock
A
A
Dittrich
H
H
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D001483:Base Sequence; D001599:Berberine; D019759:Carbon-Nitrogen Lyases; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers; D008969:Molecular Sequence Data; D009036:Moths; D010089:Oxidoreductases, N-Demethylating / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D014166:Transferases / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification
D017931:DNA Primers; D010940:Plant Proteins; D001599:Berberine; C071690:reticuline oxidase; D010089:Oxidoreductases, N-Demethylating; D014166:Transferases; D019759:Carbon-Nitrogen Lyases; C022684:strictosidine synthetase