Henke Nadja A, Wichmann Julian, Baier Thomas, Frohwitter Jonas, Lauersen Kyle J, Risse Joe M, Peters-Wendisch Petra, Kruse Olaf, Wendisch Volker F
Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, D-33615 Bielefeld, Germany.
Algae Biotechnology & Bioenergy, Faculty of Biology & CeBiTec, Bielefeld University, D-33615 Bielefeld, Germany.
Genes (Basel). 2018 Apr 17;9(4):219. doi: 10.3390/genes9040219.
Patchoulol is a sesquiterpene alcohol and an important natural product for the perfume industry. is the prominent host for the fermentative production of amino acids with an average annual production volume of ~6 million tons. Due to its robustness and well established large-scale fermentation, has been engineered for the production of a number of value-added compounds including terpenoids. Both C40 and C50 carotenoids, including the industrially relevant astaxanthin, and short-chain terpenes such as the sesquiterpene valencene can be produced with this organism. In this study, systematic metabolic engineering enabled construction of a patchoulol producing strain by applying the following strategies: (i) construction of a farnesyl pyrophosphate-producing platform strain by combining genomic deletions with heterologous expression of from ; (ii) prevention of carotenoid-like byproduct formation; (iii) overproduction of limiting enzymes from the 2-c-methyl-d-erythritol 4-phosphate (MEP)-pathway to increase precursor supply; and (iv) heterologous expression of the plant patchoulol synthase gene PS from . Additionally, a proof of principle liter-scale fermentation with a two-phase organic overlay-culture medium system for terpenoid capture was performed. To the best of our knowledge, the patchoulol titers demonstrated here are the highest reported to date with up to 60 mg L and volumetric productivities of up to 18 mg L d.
广藿香醇是一种倍半萜醇，是香料工业中一种重要的天然产物。[具体生物名称]是氨基酸发酵生产的主要宿主，年平均产量约600万吨。由于其具有较强的适应性且大规模发酵技术成熟，已对其进行工程改造以生产多种增值化合物，包括萜类化合物。利用这种生物可以生产C40和C50类胡萝卜素，包括具有工业相关性的虾青素，以及短链萜类化合物，如倍半萜瓦伦西亚烯。在本研究中，通过应用以下策略，系统代谢工程实现了构建产广藿香醇的[具体生物名称]菌株：（i）通过将基因组缺失与来自[具体生物名称]的[相关基因名称]的异源表达相结合，构建一个生产法呢基焦磷酸的平台菌株；（ii）防止类胡萝卜素样副产物的形成；（iii）过量表达来自2-C-甲基-D-赤藓糖醇-4-磷酸（MEP）途径的限制酶以增加前体供应；（iv）异源表达来自[具体植物名称]的植物广藿香醇合酶基因PS。此外，还进行了原理验证的升规模发酵，采用两相有机覆盖培养基系统进行萜类化合物捕获。据我们所知，此处展示的广藿香醇滴度是迄今为止报道的最高值，高达60 mg/L，体积产率高达18 mg/(L·d)。
