Microbial production of plant benzylisoquinoline alkaloids
Microbial production of plant benzylisoquinoline alkaloids
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DOI：
10.1073/pnas.0802981105
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发表时间：
2008-05-27
影响因子：
11.1
通讯作者：
Sato, Fumihiko
中科院分区：
文献类型：
--
作者：
Minami, Hiromichi;Kim, Ju-Sung;Sato, Fumihiko
苄基异喹啉生物碱，如镇痛化合物吗啡和可待因，以及抗菌剂小檗碱、巴马汀和木兰碱，是从罂粟科、小檗科、毛茛科、木兰科和许多其他植物科的酪氨酸合成的。生物碱在植物次生代谢的严格控制下难以大规模生产，而且它们过于复杂，无法进行成本有效的化学合成。利用微生物酶和植物酶相结合的方法，以多巴胺为原料，利用转基因大肠杆菌粗酶合成苄基异喹啉生物碱的关键中间体（S）-牛心果碱。在1h内（S）-牛心果碱的最终产量为55 mg/L。此外，我们利用转基因大肠杆菌的不同组合培养物，通过牛心果碱从多巴胺合成了阿朴啡生物碱木兰花碱或原小檗碱生物碱臭草碱。大肠杆菌和酿酒酵母细胞。木兰花碱和臭牡丹碱的最终产量分别为7.2和8.3毫克/升培养基。这些结果表明，将植物基因的微生物系统，不仅能够大规模生产稀缺的苄基异喹啉生物碱，但也可能开辟新的苄基异喹啉生物碱的生产途径。
Benzylisoquinoline alkaloids, such as the analgesic compounds morphine and codeine, and the antibacterial agents berberine, palmatine, and magnoflorine, are synthesized from tyrosine in the Papaveraceae, Berberidaceae, Ranunculaceae, Magnoliaceae, and many other plant families. It is difficult to produce alkaloids on a large scale under the strict control of secondary metabolism in plants, and they are too complex for cost-effective chemical synthesis. By using a system that combines microbial and plant enzymes to produce desired benzylisoquinoline alkaloids, we synthesized (S)-reticuline, the key intermediate in benzylisoquinoline alkaloid biosynthesis, from dopamine by crude enzymes from transgenic Escherichia coli. The final yield of (S)-reticuline was 55 mg/liter within 1 h. Furthermore, we synthesized an aporphine alkaloid, magnoflorine, or a protoberberine alkaloid, scoulerine, from dopamine via reticuline by using different combination cultures of transgenic E. coli and Saccharomyces cerevisiae cells. The final yields of magnoflorine and scoulerine were 7.2 and 8.3 mg/liter culture medium. These results indicate that microbial systems that incorporate plant genes cannot only enable the mass production of scarce benzylisoquinoline alkaloids but may also open up pathways for the production of novel benzylisoquinoline alkaloids.