Heterologous biosynthesis of triterpenoid ambrein in engineered Escherichia coli

Ke, Di; Caiyin, Qinggele; Zhao, Fanglong; Liu, Ting; Lu, Wenyu

doi:10.1007/s10529-017-2483-2

Cited by 15 publications

(10 citation statements)

References 15 publications

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“…Upscaling to 5 L bioreactors resulted in over 100 mg L −1 of (+)-ambrein, underlining the potential of this engineered P. pastoris strain as triterpenoid production platform. Combining cell and enzyme engineering approaches, (+)-ambrein yields of our strain clearly exceed production levels of the only other whole-cell system, E. coli , that had been reported so far ( Ke et al, 2018 ). Metabolic and enzyme engineering approaches as well as adjusting cultivation and process conditions are highly promising and will further enhance (+)-ambrein yield.…”

Section: Discussionmentioning

confidence: 64%

“…Our work confirmed the findings of Ueda et al (2013) who stated that these two compounds cannot be separated easily due to their highly similar chemical structure. It should not go unnoticed that contradicting work was published recently ( Ke et al, 2018 ). Consequently, a GC-MS method suitable to separate highly similar compounds such as squalene/3-deoxyachilleol and 8α-hydroxypolypoda-13,17,21-triene/(+)-ambrein was of utmost importance and therefore established ( Fig.…”

Section: Resultsmentioning

confidence: 95%

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Whole-cell (+)-ambrein production in the yeast Pichia pastoris

Moser

Strohmeier

Leitner

et al. 2018

Metabolic Engineering Communications

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The triterpenoid (+)-ambrein is a natural precursor for (-)-ambrox, which constitutes one of the most sought-after fragrances and fixatives for the perfume industry. (+)-Ambrein is a major component of ambergris, an intestinal excretion of sperm whales that is found only serendipitously. Thus, the demand for (-)-ambrox is currently mainly met by chemical synthesis. A recent study described for the first time the applicability of an enzyme cascade consisting of two terpene cyclases, namely squalene-hopene cyclase from Alicyclobacillus acidocaldarius (AaSHC D377C) and tetraprenyl-β-curcumene cyclase from Bacillus megaterium (BmeTC) for in vitro (+)-ambrein production starting from squalene. Yeasts, such as Pichia pastoris, are natural producers of squalene and have already been shown in the past to be excellent hosts for the biosynthesis of hydrophobic compounds such as terpenoids. By targeting a central enzyme in the sterol biosynthesis pathway, squalene epoxidase Erg1, intracellular squalene levels in P. pastoris could be strongly enhanced. Heterologous expression of AaSHC D377C and BmeTC and, particularly, development of suitable methods to analyze all products of the engineered strain provided conclusive evidence of whole-cell (+)-ambrein production. Engineering of BmeTC led to a remarkable one-enzyme system that was by far superior to the cascade, thereby increasing (+)-ambrein levels approximately 7-fold in shake flask cultivation. Finally, upscaling to 5 L bioreactor yielded more than 100 mg L−1 of (+)-ambrein, demonstrating that metabolically engineered yeast P. pastoris represents a valuable, whole-cell system for high-level production of (+)-ambrein.

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Section: Discussionmentioning

confidence: 64%

Section: Resultsmentioning

confidence: 95%

Whole-cell (+)-ambrein production in the yeast Pichia pastoris

Moser

Strohmeier

Leitner

et al. 2018

Metabolic Engineering Communications

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“…2c). In a bioreactor, the production of 1 by BmeTC D373C in yeast Pichia pastoris reached a maximum titer of 105 mg L -1 of culture medium 17 , which was higher than that produced using other host systems or with the two enzyme system involving SHC D377C and BmeTC WT (Supplementary Table 1) [17][18][19] .…”

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confidence: 88%

Construction of an artificial system for ambrein biosynthesis and investigation of some biological activities of ambrein

Yamabe¹,

Kawagoe²,

Okuno³

et al. 2020

Sci Rep

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Ambergris, a sperm whale metabolite, has long been used as a fragrance and traditional medication, but it is now rarely available. The odor components of ambergris result from the photooxidative degradation of the major component, ambrein. The pharmacological activities of ambergris have also been attributed to ambrein. However, efficient production of ambrein and odor compounds has not been achieved. Here, we constructed a system for the synthesis of ambrein and odor components. First, we created a new triterpene synthase, “ambrein synthase,” for mass production of ambrein by redesigning a bacterial enzyme. The ambrein yields were approximately 20 times greater than those reported previously. Next, an efficient photooxidative conversion system from ambrein to a range of volatiles of ambergris was established. The yield of volatiles was 8–15%. Finally, two biological activities, promotion of osteoclast differentiation and prevention of amyloid β-induced apoptosis, were discovered using the synthesized ambrein.

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“…41 In addition, the introduction of SQS into E. coli can also provide a central precursor for the synthesis of other cyclic triterpenoids, which will facilitate the overproduction of squalene. 42 Key ratelimiting enzymes in MVA pathway include isopentenyl diphosphate isomerase (IDI), farnesyl pyrophosphate synthase (ISPA/ERG20) and HMG-CoA reductase (HMGR). When squalene synthase genes from green alga of Botryococcus braunii were introduced into Synechocytis sp.…”

Section: Regulation Of Rate-limiting Enzymes Expressionmentioning

confidence: 99%

Advanced Strategies for the Efficient Production of Squalene by Microbial Fermentation

Wang

et al. 2022

Ind. Eng. Chem. Res.

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Squalene is an isoprene compound, which shows excellent properties, such as antioxidant and anticancer and so on. Currently, squalene has been widely used in medicine, cosmetics, vaccine adjuvants, and other fields. The traditional methods for squalene production mainly include natural extraction and chemical synthesis, which cannot meet the increasing market demand. Squalene production through microbial fermentation has become a promising alternative owing to its high efficiency and environmental friendliness. To improve the squalene production, a series of strategies including metabolic engineering and fermentation optimization and so on have been recruited. Accordingly, this review will comprehensively discuss various strategies including the isolation of high squalene producer, metabolic regulation of synthetic pathway and process optimization to improve the synthetic efficiency of squalene. Challenges and future perspectives for microbial squalene synthesis were also proposed.

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Heterologous biosynthesis of triterpenoid ambrein in engineered Escherichia coli

Abstract: An E. coli chassis for ambrein production was constructed by combining the squalene synthesis module with the downstream cyclization module.

Cited by 15 publications

References 15 publications

Whole-cell (+)-ambrein production in the yeast Pichia pastoris

Whole-cell (+)-ambrein production in the yeast Pichia pastoris

Construction of an artificial system for ambrein biosynthesis and investigation of some biological activities of ambrein

Advanced Strategies for the Efficient Production of Squalene by Microbial Fermentation

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