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利用代谢工程改造的谷氨酸棒杆菌从葡萄糖生物合成蒎烯。

Biosynthesis of pinene from glucose using metabolically-engineered Corynebacterium glutamicum.

作者信息

Kang Min-Kyoung, Eom Jin-Hee, Kim Yunje, Um Youngsoon, Woo Han Min

机构信息

Clean Energy Research Center, Korea Institute of Science and Technology, Hwarangno 14-gil 5 Seongbuk-gu, Seoul, 136-791, Republic of Korea.

出版信息

Biotechnol Lett. 2014 Oct;36(10):2069-77. doi: 10.1007/s10529-014-1578-2. Epub 2014 Jun 15.

Abstract

Pinene is a monoterpenes (C10) that is produced in a genetically-engineered microbial host for its industrial applications in fragrances, flavoring agents, pharmaceuticals, and biofuels. Herein, we have metabolically-engineered Corynebacterium glutamicum, to produce pinene and studied its toxicity in C. glutamicum. Geranyl diphosphate synthases (GPPS) and pinene synthases (PS), obtained from Pinus taeda and Abies grandis, were co-expressed with over-expressed native 1-deoxy-d-xylulose-5-phosphate synthase (Dxs) and isopentenyl diphosphate isomerase (Idi) from C. glutamicum using CoryneBrick vector. Most strains expressing PS-GPPSs produced detectable amounts of pinene, but co-expression of DXS and IDI with PS (P. taeda) and GPPS (A. grandis) resulted in 27 μg ± 7 α-pinene g(-1) cell dry weight, which is the first report in C. glutamicum. Further engineering of PS and GPPS in the C. glutamicum strain may increase pinene production.

摘要

蒎烯是一种单萜(C10),可在基因工程改造的微生物宿主中产生,用于香料、调味剂、药品和生物燃料等工业应用。在此,我们对谷氨酸棒杆菌进行了代谢工程改造以生产蒎烯,并研究了其在谷氨酸棒杆菌中的毒性。从火炬松和巨冷杉中获得的香叶基二磷酸合酶(GPPS)和蒎烯合酶(PS),与谷氨酸棒杆菌中过表达的天然1-脱氧-D-木酮糖-5-磷酸合酶(Dxs)和异戊烯基二磷酸异构酶(Idi)一起,使用CoryneBrick载体共表达。大多数表达PS-GPPSs的菌株都产生了可检测量的蒎烯,但DXS和IDI与PS(火炬松)和GPPS(巨冷杉)的共表达产生了27 μg ± 7 α-蒎烯/克细胞干重,这是谷氨酸棒杆菌中的首次报道。在谷氨酸棒杆菌菌株中对PS和GPPS进行进一步工程改造可能会提高蒎烯产量。

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