Kutchan T M, Bock A, Dittrich H
Laboratorium für Molekulare Biologie, Universität München, Germany.
Phytochemistry. 1994 Jan;35(2):353-60. doi: 10.1016/s0031-9422(00)94763-0.
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.
利用杆状病毒表达系统，已在秋粘虫（Spodoptera frugiperda）的细胞培养物（Sf9）中实现了编码生物碱生物合成酶的cDNA的异源表达，这些酶包括来自蛇根木（Rauvolfia serpentina）的裂环马钱子苷合酶[EC 4.3.3.2]和来自加利福尼亚罂粟（Eschscholtzia californica）的小檗碱桥酶[（S）-网状番荔枝碱：氧氧化还原酶（亚甲基桥形成），EC 1.5.3.9]。该表达导致每种植物酶以催化活性形式过量产生。对于裂环马钱子苷合酶和小檗碱桥酶，每升细胞培养物获得的最大产量为4毫克纯化的活性酶。
