Metabolic engineering for resveratrol derivative biosynthesis in Escherichia coli.
Metabolic engineering for resveratrol derivative biosynthesis in Escherichia coli.
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DOI：
10.14348/molcells.2015.2188
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发表时间：
2015-04
影响因子：
3.8
通讯作者：
Kim CY
中科院分区：
文献类型：
--
作者：
Jeong YJ;Woo SG;An CH;Jeong HJ;Hong YS;Kim YM;Ryu YB;Rho MC;Lee WS;Kim CY
我们先前报道了SbROMT 3syn重组蛋白通过在重组E.杆菌为进一步研究E.通过在大肠杆菌中表达参与芪生物合成的酶，我们从大黄、花生和葡萄中分离了三个芪合酶（STS）基因，从葡萄和高粱中分离了两个白藜芦醇O-甲基转移酶（ROMT）基因。RpSTS在重组E. coli与其它AhSTS和VrSTS基因进行了比较。在三个STS中，只有AhSTS能够从对香豆酸生产白藜芦醇。因此，为了提高RpSTS、VrROMT和SbROMT 3在大肠杆菌中的溶解度。在大肠杆菌中，我们合成了RpSTS、VrROMT和SbROMT 3基因，并在密码子优化后表达了一个或两个基因以及天蓝色链霉菌的肉桂酸/4-香豆酸：辅酶A连接酶（CCL）基因。我们的HPLC和LC-MS分析表明，重组E。当对香豆酸作为前体时，同时表达ScCCL和RpSTSsyn的大肠杆菌导致白藜芦醇的产生。此外，SbROMT 3syn在重组E.大肠杆菌细胞产生白藜芦醇及其单甲基化衍生物松芪，作为对香豆酸的主要产物。然而，非常少量的紫檀芪仅在重组E.表达ScCCL、RpSTSsyn和SbROMT 3syn基因的大肠杆菌细胞。这些结果表明，RpSTSsyn在E. coli细胞。
We previously reported that the SbROMT3syn recombinant protein catalyzes the production of the methylated resveratrol derivatives pinostilbene and pterostilbene by methylating substrate resveratrol in recombinant E. coli. To further study the production of stilbene compounds in E. coli by the expression of enzymes involved in stilbene biosynthesis, we isolated three stilbene synthase (STS) genes from rhubarb, peanut, and grape as well as two resveratrol O-methyltransferase (ROMT) genes from grape and sorghum. The ability of RpSTS to produce resveratrol in recombinant E. coli was compared with other AhSTS and VrSTS genes. Out of three STS, only AhSTS was able to produce resveratrol from p-coumaric acid. Thus, to improve the solubility of RpSTS, VrROMT, and SbROMT3 in E. coli, we synthesized the RpSTS, VrROMT and SbROMT3 genes following codon-optimization and expressed one or both genes together with the cinnamate/4-coumarate:coenzyme A ligase (CCL) gene from Streptomyces coelicolor. Our HPLC and LC-MS analyses showed that recombinant E. coli expressing both ScCCL and RpSTSsyn led to the production of resveratrol when p-coumaric acid was used as the precursor. In addition, incorporation of SbROMT3syn in recombinant E. coli cells produced resveratrol and its mono-methylated derivative, pinostilbene, as the major products from p-coumaric acid. However, very small amounts of pterostilbene were only detectable in the recombinant E. coli cells expressing the ScCCL, RpSTSsyn and SbROMT3syn genes. These results suggest that RpSTSsyn exhibits an enhanced enzyme activity to produce resveratrol and SbROMT3syn catalyzes the methylation of resveratrol to produce pinostilbene in E. coli cells.
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影响因子：
6.7
作者：
Lee, Seung Woong;Hwang, Byung Soon;Rho, Mun-Chual
通讯作者：
Rho, Mun-Chual
影响因子：
7.2
作者：
Remsberg, Connie M.;Yanez, Jaime A.;Davies, Neal M.
通讯作者：
Davies, Neal M.
影响因子：
3.2
作者：
Becker, JVW;Armstrong, GO;Pretorius, IS
通讯作者：
Pretorius, IS
影响因子：
3.5
作者：
MELCHIOR, F;KINDL, H
通讯作者：
KINDL, H
影响因子：
3.5
作者：
Watts, Kevin T;Lee, Pyung C;Schmidt-Dannert, Claudia
通讯作者：
Schmidt-Dannert, Claudia