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Genes 2018, 9(4), 219; https://doi.org/10.3390/genes9040219

Patchoulol Production with Metabolically Engineered Corynebacterium glutamicum

1
Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, D-33615 Bielefeld, Germany
2
Algae Biotechnology & Bioenergy, Faculty of Biology & CeBiTec, Bielefeld University, D-33615 Bielefeld, Germany
3
Fermentation Technology, Technical Faculty & CeBiTec, Bielefeld University, D-33615 Bielefeld, Germany
*
Author to whom correspondence should be addressed.
Received: 26 March 2018 / Revised: 10 April 2018 / Accepted: 16 April 2018 / Published: 17 April 2018
(This article belongs to the Special Issue Microbial Metabolic Engineering)
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Abstract

Patchoulol is a sesquiterpene alcohol and an important natural product for the perfume industry. Corynebacterium glutamicum 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, C. glutamicum 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 C. glutamicum strain by applying the following strategies: (i) construction of a farnesyl pyrophosphate-producing platform strain by combining genomic deletions with heterologous expression of ispA from Escherichia coli; (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 PcPS from Pogostemon cablin. 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−1 and volumetric productivities of up to 18 mg L−1 d−1. View Full-Text
Keywords: patchoulol; Corynebacterium glutamicum; sesquiterpene; metabolic engineering; algae; Escherichia coli patchoulol; Corynebacterium glutamicum; sesquiterpene; metabolic engineering; algae; Escherichia coli
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Henke, N.A.; Wichmann, J.; Baier, T.; Frohwitter, J.; Lauersen, K.J.; Risse, J.M.; Peters-Wendisch, P.; Kruse, O.; Wendisch, V.F. Patchoulol Production with Metabolically Engineered Corynebacterium glutamicum. Genes 2018, 9, 219.

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