Production of sesquiterpenoid zerumbone from metabolic engineered Saccharomyces cerevisiae

Zhang, Chuanbo; Liu, Jingjing; Zhao, Fanglong; Lu, Chunzhe; Zhao, Guang‐Rong; Lu, Wenyu

doi:10.1016/j.ymben.2018.07.010

Cited by 58 publications

(60 citation statements)

References 37 publications

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“…Firstly, FPP supply was optimized to improve the production of 13 R -MO. CfTPS2 and CfTPS3 were introduced into the well-established chassis strain LW03 ( tHMG1 and ERG20 overexpressed and the ERG9 promoter replaced by MET3p ), which was constructed for the production of the sesquiterpene α-humulene in our previous study, resulting in strain LZJ1 [18]. LZJ1 could produce 2.86 mg/L of 13 R -MO, which was slightly higher than that produced by the original strain GW-1.…”

Section: Resultsmentioning

confidence: 99%

“…In our previous study, the ERG9 promoter was replaced with MET3p in S. cerevisiae to produce levopimaradiene, a type of diterpene produced by plants at a titer 8.5-fold higher than that produced by its parental strain. However, this strategy needs the addition of methionine, which could be metabolized by the cell [18, 29]. In this study, a well-established sesquiterpene-producing yeast platform, LW03, was used to produce 13 R -MO by overexpressing CfTPS2 and CfTPS3 , but the titer of 13 R -MO did not increase significantly.…”

Section: Discussionmentioning

confidence: 99%

“…The feed solution was prepared according to our previously reported methods and was added automatically according to the pH and DO after 18 h [18]. When the pH was above 5.5 and DO was above 35%, the feed solution was added at a rate of 2 mL/min.…”

Section: Methodsmentioning

confidence: 99%

“…A yeast strain that produces high titers (40 g/L) of amorphadiene has demonstrated potential for natural terpenoid production [16]; however, the yields of diterpene were markedly lower than that of sesquiterpenoids. Metabolic engineering to increase the FPP pool, such as increasing precursor levels, inhibiting the ergosterol synthetic pathway, and deleting branch pathways, was shown to remarkably increased sesquiterpenoid production [17, 18]. However, S. cerevisiae GGPP synthase (Bts1p) showed a higher K m value (3.2 μM) for FPP than for squalene synthase (2.5 μM) [19].…”

Section: Introductionmentioning

confidence: 99%

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High-titer production of 13R-manoyl oxide in metabolically engineered Saccharomyces cerevisiae

Zhang

et al. 2019

Microb Cell Fact

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Background Diterpenoids are a large class of natural products with complex structures and broad commercial applications as food additives, important medicines, and fragrances. However, their low abundance in plants and high structural complexity limit their applications. Therefore, it is important to create an efficient diterpenoid-producing yeast cell factory of the production of various high-value diterpenoid compounds in a cost-effective manner Results In this study, 13 R -manoyl oxide (13 R -MO; 2.31 mg/L) was produced by expressing CfTPS2 and CfTPS3 from Coleus forskohlii in Saccharomyces cerevisiae . The 13 R -MO titer was increased by 142-fold to 328.15 mg/L via the stepwise metabolic engineering of the original strain, including the overexpression of the rate-limiting genes ( tHMG1 and ERG20 ) of the mevalonate pathway, transcription and protein level regulation of ERG9 , Bts1p and Erg20 F96C p fusion, and the overexpression of tCfTPS2 and tCfTPS3 (excision of the N-terminal plastid transit peptide sequences of CfTPS2 and CfTPS3 ). The final titer of 13 R -MO reached up to 3 g/L by fed-batch fermentation in a 5 L bioreactor. Conclusions In this study, an efficient 13 R -MO yeast cell factory was constructed, which achieved the de novo production of 3 g/L of 13R-MO from glucose. To the best of our knowledge, this is the highest 13 R -MO titer reported to date. Furthermore, the metabolic engineering strategies presented here could be used to produce other valuable diterpenoid compounds in yeast. Electronic supplementary material The online version of this article (10.1186/s12934-019-1123-z) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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High-titer production of 13R-manoyl oxide in metabolically engineered Saccharomyces cerevisiae

Zhang

et al. 2019

Microb Cell Fact

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“…Cytochromes P450 (P450s) play a crucial role in yielding final terpenoid products with wide chemical diversity and bioactivities. S. cerevisiae is a favored host for expressing P450s on account of advanced protein expression mechanism, abundant intracellular membranes and the inherent benefits of large-scale microbial fermentation (Paddon et al, 2013;Gold et al, 2018;Wong et al, 2018;Zhang et al, 2018a). It is generally deemed that the ability to express soluble P450s in E. coli is limited.…”

Section: Cytochromes P450 and Post-modifications Of Terpenoidsmentioning

confidence: 99%

Production of Terpenoids by Synthetic Biology Approaches

Zhang

Hong

2020

Front. Bioeng. Biotechnol.

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Terpenoids are a large family of natural products with remarkable diverse biological functions, and have a wide range of applications as pharmaceuticals, flavors, pigments, and biofuels. Synthetic biology is presenting possibilities for sustainable and efficient production of high value-added terpenoids in engineered microbial cell factories, using Escherichia coli and Saccharomyces cerevisiae which are identified as well-known industrial workhorses. They also provide a promising alternative to produce non-native terpenes on account of available genetic tools in metabolic engineering and genome editing. In this review, we summarize the recent development in terpenoids production by synthetic biology approaches.

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