Whole-Cell Production of Patchouli Oil Sesquiterpenes in Escherichia coli: Metabolic Engineering and Fermentation Optimization in Solid-Liquid Phase Partitioning Cultivation.
Whole-Cell Production of Patchouli Oil Sesquiterpenes in Escherichia coli: Metabolic Engineering and Fermentation Optimization in Solid-Liquid Phase Partitioning Cultivation.
复制标题
大肠杆菌全细胞生产广藿香油倍半萜：固-液相分配培养中的代谢工程和发酵优化。
DOI：
10.1021/acsomega.0c04590
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发表时间：
2020-12-22
期刊：
影响因子：
4.1
通讯作者：
Beutel S
中科院分区：
文献类型：
--
作者：
Aguilar F;Ekramzadeh K;Scheper T;Beutel S
广藿香油是香水的主要成分，由于其主要成分（−）-广藿香醇而赋予黑暗的木质气味。对广藿香油日益增长的需求引起了人们对开发生物技术过程以确保可靠供应的兴趣。在此，我们报告的生产广藿香油倍半萜代谢工程大肠杆菌菌株，使用固-液相分配培养。（-）-广藿香醇的生产是可能的，使用内源性甲基磷酸途径和过表达广藿香的（-）-广藿香醇合酶同种型，但在低滴度。为了提高（-）-百曲醇的产量，在多质粒PTS + Mev菌株中过表达外源甲羟戊酸途径，这使（-）-百曲醇滴度增加了5倍。通过评估几种确定的培养基，并优化培养液的pH和温度，进一步改善发酵，将（-）-百秋醇滴度提高3倍。为了增加从发酵中的（-）-百秋醇回收率，通过筛选聚合物吸附器来分析固-液相分配培养，其中Diaion HP 20吸附器从所有测试中证明了最高的（-）-百秋醇回收率。将发酵放大到补料分批生物反应器，达到40.2 mg L-1的（-）-百曲醇滴度和20.1 mg L-1 d-1的生产率。由PTS + Mev菌株产生的萜烯特征和香味类似于广藿香油，包含（-）-广藿香醇作为主要产物，以及α-布来烯、反式-β-香芹烯、β-广藿香烯和愈创木-5，11-二烯作为副产物。本研究首次对E.该研究为开发微生物生产芳香萜类化合物的可持续生物过程提供了新的思路。
Patchouli oil is a major ingredient in perfumery, granting a dark-woody scent due to its main constituent (−)-patchoulol. The growing demand for patchouli oil has raised interest in the development of a biotechnological process to assure a reliable supply. Herein, we report the production of patchouli oil sesquiterpenes by metabolically engineered Escherichia coli strains, using solid–liquid phase partitioning cultivation. The (−)-patchoulol production was possible using the endogenous methylerythritol phosphate pathway and overexpressing a (−)-patchoulol synthase isoform from Pogostemon cablin but at low titers. To improve the (−)-patchoulol production, the exogenous mevalonate pathway was overexpressed in the multi-plasmid PTS + Mev strain, which increased the (−)-patchoulol titer 5-fold. Fermentation was improved further by evaluating several defined media, and optimizing the pH and temperature of culture broth, enhancing the (−)-patchoulol titer 3-fold. To augment the (−)-patchoulol recovery from fermentation, the solid–liquid phase partitioning cultivation was analyzed by screening polymeric adsorbers, where the Diaion HP20 adsorber demonstrated the highest (−)-patchoulol recovery from all tests. Fermentation was scaled-up to fed-batch bioreactors, reaching a (−)-patchoulol titer of 40.2 mg L–1 and productivity of 20.1 mg L–1 d–1. The terpene profile and aroma produced from the PTS + Mev strain were similar to the patchouli oil, comprising (−)-patchoulol as the main product, and α-bulnesene, trans-β-caryophyllene, β-patchoulene, and guaia-5,11-diene as side products. This investigation represents the first study of (−)-patchoulol production in E. coli by solid–liquid phase partitioning cultivation, which provides new insights for the development of sustainable bioprocesses for the microbial production of fragrant terpenes.
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影响因子：
2.9
作者：
Guan, ZQ;Breazeale, SD;Raetz, CRH
通讯作者：
Raetz, CRH
影响因子：
4
作者：
Chubukov V;Desmarais JJ;Wang G;Chan LJG;Baidoo EE;Petzold CJ;Keasling JD;Mukhopadhyay A
通讯作者：
Mukhopadhyay A
影响因子：
6.4
作者：
Li Z;Carstensen B;Rinas U
通讯作者：
Rinas U
影响因子：
8.4
作者：
Asadollahi, Mohammad A.;Maury, Jerome;Nielsen, Jens
通讯作者：
Nielsen, Jens
影响因子：
4.6
作者：
Cao, Yujin;Zhang, Rubing;Liu, Huizhou
通讯作者：
Liu, Huizhou