Advances in the metabolic engineering of Yarrowia lipolytica for the production of terpenoids

Ma, Yi-Rong; Wang, Kaifeng; Wang, Weijian; Ding, Ying; Shi, Tian‐Qiong; Huang, He; Ji, Xiao‐Jun

doi:10.1016/j.biortech.2019.02.116

Cited by 89 publications

(79 citation statements)

References 73 publications

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“…Several useful synthetic biology tools have been aiding the creation of new and better performing Y. lipolytica strains [42,43]. To date, the highest yields of lipid, lycopene and carotene produced via microbial cell factories have been achieved by using engineered Y. lipolytica strains [7,12].…”

Section: Discussionmentioning

confidence: 99%

“…The use of generally recognized as safe (GRAS) microorganisms to convert natural raw materials into products leads to aroma compounds that can be described as natural, if these compounds are known to be components of the natural raw materials, according to the United States and the European Union flavor regulations [9]. As a consequence, companies and researchers have shifted their focus from Escherichia coli (E. coli) to GRAS microorganisms, such as yeasts, for terpenoids production [10][11][12][13].…”

Section: Introductionmentioning

confidence: 99%

“…Recently, the GRAS, unconventional yeast Y. lipolytica has rapidly emerged as a valuable cell factory for the production of terpenoids due to its own endogenous mevalonate pathway (MVA) and its high lipid production capacity [7,12]. Metabolic engineering and synthetic biology offer the ability of rewiring microbial carbon fluxes to create efficient cell factories for the production of natural products.…”

Section: Introductionmentioning

confidence: 99%

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A modular pathway engineering strategy for the high-level production of β-ionone in Yarrowia lipolytica

Yang

Lin

et al. 2020

Microb Cell Fact

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Background: The GRAS and oleaginous yeast Yarrowia lipolytica (Y. lipolytica) is an attractive cell factory for the production of chemicals and biofuels. The production of many natural products of commercial interest have been investigated in this cell factory by introducing heterologous biosynthetic pathways and by modifying the endogenous pathways. However, since natural products anabolism involves long pathways and complex regulation, re-channelling carbon into the product of target compounds is still a cumbersome work, and often resulting in low production performance. Results: In this work, the carotenogenic genes contained carB and bi-functional carRP from Mucor circinelloides and carotenoid cleavage dioxygenase 1 (CCD1) from Petunia hybrida were introduced to Y. lipolytica and led to the low production of β-ionone of 3.5 mg/L. To further improve the β-ionone synthesis, we implemented a modular engineering strategy for the construction and optimization of a biosynthetic pathway for the overproduction of β-ionone in Y. lipolytica. The strategy involved the enhancement of the cytosolic acetyl-CoA supply and the increase of MVA pathway flux, yielding a β-ionone titer of 358 mg/L in shake-flask fermentation and approximately 1 g/L (~ 280-fold higher than the baseline strain) in fed-batch fermentation. Conclusions: An efficient β-ionone producing GRAS Y. lipolytica platform was constructed by combining integrated overexpressed of heterologous and native genes. A modular engineering strategy involved the optimization pathway and fermentation condition was investigated in the engineered strain and the highest β-ionone titer reported to date by a cell factory was achieved. This effective strategy can be adapted to enhance the biosynthesis of other terpenoids in Y. lipolytica.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A modular pathway engineering strategy for the high-level production of β-ionone in Yarrowia lipolytica

Yang

Lin

et al. 2020

Microb Cell Fact

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“…production NADPH as the primary biological reducing equivalent protects cell from oxidative stress and extend carbon-carbon backbones, which was also reported as the major rate-limiting precursor in fatty acids synthesis in oleaginous species Wasylenko, Ahn, & Stephanopoulos, 2015). HMG-CoA reductase (HMGR) is the first rate-limiting enzyme in the mevalonate pathway and plays critical role in regulating squalene biosynthesis (Ma et al, 2019).…”

Section: Augmenting Napdh and Acetyl-coa Precursor Pathways To Improvmentioning

confidence: 99%

Optimizing mevalonate pathway for squalene production inYarrowia lipolytica

Liu

Wang

Deng

et al. 2020

Preprint

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Li Deng) and pengxu@umbc.edu (PX). AbstractSqualene is the gateway molecule for triterpene-based natural products and steroidsbased pharmaceuticals. As a super lubricant, it has been used widely in health care industry due to its skin compatibility and thermostability. Squalene is traditionally sourced from sharkhunting or oil plant extraction, which is cost-prohibitive and not sustainable. Reconstitution of squalene biosynthetic pathway in microbial hosts is considered as a promising alternative for cost-efficient and scalable synthesis of squalene. In this work, we reported the engineering of the oleaginous yeast, Y. lipolytica, as a potential host for squalene production. We systematically identified the bottleneck of the pathway and discovered that the native HMG-CoA reductase led to the highest squalene improvement. With the recycling of NADPH from the mannitol cycle, the engineered strain produced about 180.3 mg/l and 188.2 mg/L squalene from glucose or acetate minimal media, respectively. By optimizing the C/N ratio, controlling the media pH and mitigating the acetyl-CoA flux competition from lipogenesis, the engineered strain produced about 502.7 mg/L squalene in shake flaks, a 28-fold increase compared to the parental strain (17.2 mg/L). We also profiled the metabolic byproducts citric acid and mannitol level and observed that they are reincorporated into cell metabolism at the late stage of fermentation. This work may serve as a baseline to harness Y. lipolytica as an oleaginous cell factory for production of squalene or terpene-based chemicals.

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“…Background β-carotene enhances the immune function of human, delays skin aging, and prevents cardiovascular diseases and cancers [1,2]. β-carotene is widely used as a nutritional supplement and antioxidant in the food and pharmaceutical industries [3,4]. The global market size of β-carotene was USD 466.7 million in 2017 [5].…”

mentioning

confidence: 99%

Enhanced β-carotene production by promoting the multivesicular body (MVB) pathway in Yarrowia lipolytica

Yang¹,

Liu²,

Shan³

et al. 2020

Preprint

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Background: β-carotene is a precursor of vitamin A and has great commercial value as an additive in foods and feeds. Many pathways not directly related to the β-carotene synthesis affect β-carotene production since the interactions among metabolic fluxes of cells confer a complex regulatory network. Engineered Y. lipolytica strain has excellent potential for β-carotene production as oleaginous yeast. Optimizing indirectly metabolic pathways in Y. lipolytica may offer a new strategy for making the β-carotene production achieve a commercially viable yield. Results:In this study, we found that the proper promotion of the multivesicular body (MVB) sorting pathway elevated the production of β-carotene by 1.58 fold when overexpressing one copy of the Did2 gene in Y. lipolytica. Through the measurement of ATP, NADPH, the mRNA, and protein level of key genes in the β-carotene synthesis pathway, the reason for β-carotene elevated was deuced that the protein level of the key enzymes (tHMG and CarA) was increased. When overexpressing two copies of the Did2 gene, the transcription level of the key genes was all elevated. However, the protein level of key enzymes in the β-carotene synthesis pathway was reduced compared with the overexpressing one copy of the Did2 gene, which resulted in reduced β-carotene content. Conclusion:This study suggests that the MVB sorting pathway is not responsible for sorting protein but has a crucial regulating effect on protein abundance in cells. Engineering the MVB sorting pathway could potentially increase the production of other high-value products. Moreover, manipulation of indirectly related metabolic pathways also is a critical strategy in synthetic biology research.

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Advances in the metabolic engineering of Yarrowia lipolytica for the production of terpenoids

Cited by 89 publications

References 73 publications

A modular pathway engineering strategy for the high-level production of β-ionone in Yarrowia lipolytica

A modular pathway engineering strategy for the high-level production of β-ionone in Yarrowia lipolytica

Optimizing mevalonate pathway for squalene production inYarrowia lipolytica

Enhanced β-carotene production by promoting the multivesicular body (MVB) pathway in Yarrowia lipolytica

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