Metabolic engineering strategies for sesquiterpene production in microorganism

Liu, Chunli; Xue, Kai; Yang, Yankun; Liu, Xiuxia; Li, Ye; Lee, Taek Soon; Bai, Zhonghu; Tan, Tianwei

doi:10.1080/07388551.2021.1924112

Cited by 32 publications

(31 citation statements)

References 212 publications

(210 reference statements)

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“…Sesquiterpenes are a large subgroup of terpenes with a wide range of applications; they have at least 121 different skeletons, including acyclic, monocyclic, bicyclic, tricyclic and tetracyclic skeletons, and have been identified in plants, bacteria, fungi and other organisms [ 1 , 2 ]. Germacrene D is a monocyclic sesquiterpene.…”

Section: Introductionmentioning

confidence: 99%

Identification of the sesquiterpene synthase AcTPS1 and high production of (–)-germacrene D in metabolically engineered Saccharomyces cerevisiae

Li¹,

Chen²,

Wan³

et al. 2022

Microb Cell Fact

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Background The sesquiterpene germacrene D is a highly promising product due to its wide variety of insecticidal activities and ability to serve as a precursor for many other sesquiterpenes. Biosynthesis of high value compounds through genome mining for synthases and metabolic engineering of microbial factories, especially Saccharomyces cerevisiae, has been proven to be an effective strategy. However, there have been no studies on the de novo synthesis of germacrene D from carbon sources in microbes. Hence, the construction of the S. cerevisiae cell factory to achieve high production of germacrene D is highly desirable. Results We identified five putative sesquiterpene synthases (AcTPS1 to AcTPS5) from Acremonium chrysogenum and the major product of AcTPS1 characterized by in vivo, in vitro reaction and NMR detection was revealed to be (–)-germacrene D. After systematically comparing twenty-one germacrene D synthases, AcTPS1 was found to generate the highest amount of (–)-germacrene D and was integrated into the terpene precursor-enhancing yeast strain, achieving 376.2 mg/L of (–)-germacrene D. Iterative engineering was performed to improve the production of (–)-germacrene D, including increasing the copy numbers of AcTPS1, tHMG1 and ERG20, and downregulating or knocking out other inhibitory factors (such as erg9, rox1, dpp1). Finally, the optimal strain LSc81 achieved 1.94 g/L (–)-germacrene D in shake-flask fermentation and 7.9 g/L (–)-germacrene D in a 5-L bioreactor, which is the highest reported (–)-germacrene D titer achieved to date. Conclusion We successfully achieved high production of (–)-germacrene D in S. cerevisiae through terpene synthase mining and metabolic engineering, providing an impressive example of microbial overproduction of high-value compounds.

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Section: Introductionmentioning

confidence: 99%

Identification of the sesquiterpene synthase AcTPS1 and high production of (–)-germacrene D in metabolically engineered Saccharomyces cerevisiae

Li¹,

Chen²,

Wan³

et al. 2022

Microb Cell Fact

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“…Terpenes, such as (+)-aristolochene and (+)-bicyclogermacrene, have great potential for applications. ,, However, large-scale biosynthesis of terpenes is very limited as a result of the poor expression of TSs in heterologous hosts, imbalanced metabolic pathways, and/or toxicity of substrates, intermediate metabolites, and terpenes. , Because the majority of studies on terpene production are focused on the optimization of precursor-providing pathways to balance metabolic pathways, decrease the toxicity of metabolites, and enhance the supply of precursors, ,, less attention has been paid to the limitations of terpene synthase itself.…”

Section: Discussionmentioning

confidence: 99%

Facile Production of (+)-Aristolochene and (+)-Bicyclogermacrene in Escherichia coli Using Newly Discovered Sesquiterpene Synthases from Penicillium expansum

Huang

et al. 2022

J. Agric. Food Chem.

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Penicillium expansum, producer of a wide array of secondary metabolites, has the potential to be a source of new terpene synthases. In this work, a platform was constructed with Escherichia coli BL21(DE3) by enhancing its endogenous 2-methyl-D-erythritol-4-phosphate pathway to supply sufficient terpenoid precursors. Using this precursor-supplying platform, we discovered two sesquiterpene synthases from P. expansum: PeTS1, a new (+)-aristolochene synthase, and PeTS4, the first microbial (+)-bicyclogermacrene synthase. To enhance the sesquiterpene production by PeTS1, we employed a MBP fusion tag to improve the heterologous protein expression, resulting in the increase of aristolochene production up to 50 mg/L in a 72 h flask culture, which is the highest production reported to date. We also realized the first biosynthesis of (+)-bicyclogermacrene, achieving 188 mg/ L in 72 h. This work highlights the great potential of this microbial platform for the discovery of new terpene synthases and opens new ways for the bioproduction of other valuable terpenoids.

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“…In recent years, numerous engineering strategies were developed to optimize terpene synthesis in different microorganisms. − This review summarizes the natural terpene biosynthesis pathways and various strategies to further improve terpene productivity and strain stability in the yeasts S. cerevisiae and Y. lipolytica using enzyme engineering, pathway engineering, and cellular engineering. Then, the future prospects of terpene production using these yeasts are discussed in light of the current progress, challenges, and trends in this field.…”

Section: Introductionmentioning

confidence: 99%

Advances in Metabolic Engineering Paving the Way for the Efficient Biosynthesis of Terpenes in Yeasts

Cui

Mai

et al. 2022

J. Agric. Food Chem.

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Terpenes are a large class of secondary metabolites with diverse structures and functions that are commonly used as valuable raw materials in food, cosmetics, and medicine. With the development of metabolic engineering and emerging synthetic biology tools, these important terpene compounds can be sustainably produced using different microbial chassis. Currently, yeasts such as Saccharomyces cerevisiae and Yarrowia lipolytica have received extensive attention as potential hosts for the production of terpenes due to their clear genetic background and endogenous mevalonate pathway. In this review, we summarize the natural terpene biosynthesis pathways and various engineering strategies, including enzyme engineering, pathway engineering, and cellular engineering, to further improve the terpene productivity and strain stability in these two widely used yeasts. In addition, the future prospects of yeast-based terpene production are discussed in light of the current progress, challenges, and trends in this field. Finally, guidelines for future studies are also emphasized.

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Metabolic engineering strategies for sesquiterpene production in microorganism

Cited by 32 publications

References 212 publications

Identification of the sesquiterpene synthase AcTPS1 and high production of (–)-germacrene D in metabolically engineered Saccharomyces cerevisiae

Identification of the sesquiterpene synthase AcTPS1 and high production of (–)-germacrene D in metabolically engineered Saccharomyces cerevisiae

Facile Production of (+)-Aristolochene and (+)-Bicyclogermacrene in Escherichia coli Using Newly Discovered Sesquiterpene Synthases from Penicillium expansum

Advances in Metabolic Engineering Paving the Way for the Efficient Biosynthesis of Terpenes in Yeasts

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