Heterologous synthesis of geranyllinalool, a diterpenol plant product, in the cyanobacterium Synechocystis
Heterologous synthesis of geranyllinalool, a diterpenol plant product, in the cyanobacterium Synechocystis
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DOI：
10.1007/s00253-016-8081-8
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发表时间：
2017-04-01
影响因子：
5
通讯作者：
Melis, Anastasios
中科院分区：
文献类型：
--
作者：
Formighieri, Cinzia;Melis, Anastasios
蓝细菌是工业上强大的光合微生物，可以通过遗传编程来合成用于家庭和工业消费的商品。在目前的工作中，集胞藻被赋予了植物次生代谢产物香叶基芳樟醇，一个商业利益的二萜醇的合成。在48小时的培养期内，每克细胞干重得到360微克香叶基芳樟醇的总平均产量。香叶基芳樟醇主要被隔离在细胞内，相当于总异源产物的60-70%，而不是像较短的异源萜烯烃那样完全渗出。香叶基芳樟醇的提取需要破坏细胞以释放和分离该化学产物。此外，香叶基芳樟醇在细胞中的积累引起了细胞健身和生物量的生长速率，这样的集胞藻转化体的复制时间比对照的1.4倍长的温和的抑制作用。发现剩余的30-40%的香叶基芳樟醇产物漂浮在密封的微生物培养物的表面上，在那里通过施加疏水覆盖层将其虹吸掉，在这种情况下不需要破坏细胞。总之，这项工作扩大了在蓝藻中异源生产萜烯和萜烯醇产品的努力，并解决了这种生产系统固有的可能性和限制。
Cyanobacteria are industrially robust photosynthetic microorganisms that can be genetically programmed to synthesize commodity products for domestic and industrial consumption. In the present work, Synechocystis was endowed with the synthesis of the plant secondary metabolite geranyllinalool, a diterpene alcohol of commercial interest. Total average yields of 360 mu g of geranyllinalool per gram of dry cell weight were obtained in the course of a 48-h cultivation period. Geranyllinalool was primarily sequestered inside the transformant cells, corresponding to 60-70% of the total heterologous product, instead of being entirely exuded, as the case is with shorter heterologous terpene hydrocarbons. Extraction of geranyllinalool necessitated disruption of the cells in order to release and isolate this chemical product. Moreover, geranyllinalool accumulation in the cells caused a mild inhibitory effect on cell fitness and biomass growth rate, such that the duplication time of Synechocystis transformants was 1.4-fold longer than that of the control. The remaining 30-40% of the geranyllinalool product was found to float on the surface of sealed transformant cultures, where it was siphoned off by applying a hydrophobic overlayer, with no need to disrupt the cells in this case. Concluding, the work extended efforts to heterologously produce terpene and terpenol products in cyanobacteria, and addressed possibilities and constrains inherent to this production system.