De novo Biosynthesis of Odd-Chain Fatty Acids in Yarrowia lipolytica Enabled by Modular Pathway Engineering

Park, Young-kyoung; Ledesma-Amaro, Rodrigo; Nicaud, Jean‐Marc

doi:10.3389/fbioe.2019.00484

Cited by 47 publications

(40 citation statements)

References 59 publications

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“…4 h). This finding is interesting for the synthesis of odd-chain fatty acids in the yeast, which relies on propionyl-CoA availability and usually requires toxic propionate supplementation or massive strain engineering [ 29 ]. In contrast, glucose resulted in a higher amount of malonyl-CoA.…”

Section: Discussionmentioning

confidence: 99%

A common approach for absolute quantification of short chain CoA thioesters in prokaryotic and eukaryotic microbes

et al. 2020

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Background: Thioesters of coenzyme A participate in 5% of all enzymatic reactions. In microbial cell factories, they function as building blocks for products of recognized commercial value, including natural products such as polyketides, polyunsaturated fatty acids, biofuels, and biopolymers. A core spectrum of approximately 5-10 short chain thioesters is present in many microbes, as inferred from their genomic repertoire. The relevance of these metabolites explains the high interest to trace and quantify them in microbial cells. Results: Here, we describe a common workflow for extraction and absolute quantification of short chain CoA thioesters in different gram-positive and gram-negative bacteria and eukaryotic yeast, i.e. Corynebacterium glutamicum, Streptomyces albus, Pseudomonas putida, and Yarrowia lipolytica. The approach assessed intracellular CoA thioesters down to the picomolar level and exhibited high precision and reproducibility for all microbes, as shown by principal component analysis. Furthermore, it provided interesting insights into microbial CoA metabolism. A succinyl-CoA synthase defective mutant of C. glutamicum exhibited an unaffected level of succinyl-CoA that indicated a complete compensation by the l-lysine pathway to bypass the disrupted TCA cycle. Methylmalonyl-CoA, an important building block of high-value polyketides, was identified as dominant CoA thioester in the actinomycete S. albus. The microbe revealed a more than 10,000-fold difference in the abundance of intracellular CoA thioesters. A recombinant strain of S. albus, which produced different derivatives of the antituberculosis polyketide pamamycin, revealed a significant depletion of CoA thioesters of the ethylmalonyl CoA pathway, influencing product level and spectrum. Conclusions: The high relevance of short chain CoA thioesters to synthetize industrial products and the interesting insights gained from the examples shown in this work, suggest analyzing these metabolites in microbial cell factories more routinely than done so far. Due to its broad application range, the developed approach appears useful to be applied this purpose. Hereby, the possibility to use one single protocol promises to facilitate automatized efforts, which rely on standardized workflows.

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Section: Discussionmentioning

confidence: 99%

A common approach for absolute quantification of short chain CoA thioesters in prokaryotic and eukaryotic microbes

et al. 2020

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“…3E). This finding is interesting for the synthesis of odd-chain fatty acids in the yeast, which relies on propionyl-CoA availability and usually requires toxic propionate supplementation or massive strain engineering [29]. In contrast, glucose resulted in a higher amount of malonyl-CoA.…”

Section: Discussionmentioning

confidence: 92%

A common approach for absolute quantification of short chain CoA thioesters in industrially relevant gram-positive and gram-negative prokaryotic and eukaryotic microbes

Gläser

Kuhl

Jovanović

et al. 2020

Preprint

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Background: Thioesters of coenzyme A participate in 5% of all enzymatic reactions and at least one third of all cellular carbon is typically metabolized through a CoA thioester. In microbial cell factories, they function as building blocks for products of recognized commercial value, including natural products such as polyketides, polyunsaturated fatty acids, biofuels, and biopolymers. A core spectrum of approximately 5 – 10 short chain thioesters is present in many microbes, as inferred from their genomic repertoire. The relevance these metabolites explains the high interest to trace and quantify them in microbial cells.Results: Here, we describe a common workflow for extraction and absolute quantification of short chain CoA thioesters in different gram-positive and gram-negative bacteria and eukaryotic yeast, i.e. Corynebacterium glutamicum, Streptomyces albus, Pseudomonas putida, and Yarrowia lipolytica. The approach detected CoA thioesters down to the level of 40 attomole and exhibited high precision and reproducibility for all microbes as shown by principal component analysis. Furthermore, it provided interesting insights into microbial CoA-spectra. A succinyl-CoA synthase defective mutant of C. glutamicum, exhibited an unaffected level of succinyl-CoA, which indicated a complete compensation of the l-lysine pathway to bypass the disrupted TCA cycle. Methylmalonyl-CoA, an important building block of high-value polyketides, was identified as dominant CoA thioester in the microbe. S. albus revealed a more than 10,000-fold difference in the abundance of intracellular CoA thioesters. A recombinant strain of S. albus, which produced different derivatives of the antituberculosis polyketide pamamycin, revealed a significant depletion of CoA thioesters of the ethylmalonyl CoA pathway, influencing product level and spectrum. Conclusions: The high relevance of short chain CoA thioesters to synthetize industrial products and the interesting insights gained from the examples shown in this work, suggest analyzing these metabolites in microbial cell factories more routinely than done so far. Due to its broad application range, the developed approach appears useful to be applied this purpose. Hereby, the possibility to use on single protocol promises to facilitate automatized efforts, which rely on standardized workflows.

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“…Recently, the deletion of the PHD1 gene and optimization of the fermentation process were applied to produce odd-chain fatty acids (mainly C 15:0 , C 17:0 , and C 17:1 ) by Y. lipolytica grown on propionate (Park et al, 2018a). Additionally, Park et al (2020) constructed an engineered Y. lipolytica capable of de novo producing odd-chain fatty acids, using glucose as sole substrate without any propionate supplementation.…”

Section: Production Of Fatty Acid-based Bioproducts Nutraceuticalsmentioning

confidence: 99%

Yarrowia lipolytica as an Oleaginous Platform for the Production of Value-Added Fatty Acid-Based Bioproducts

et al. 2021

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The microbial fermentation process has been used as an alternative pathway to the production of value-added natural products. Of the microorganisms, Yarrowia lipolytica, as an oleaginous platform, is able to produce fatty acid-derived biofuels and biochemicals. Nowadays, there are growing progresses on the production of value-added fatty acid-based bioproducts in Y. lipolytica. However, there are fewer reviews performing the metabolic engineering strategies and summarizing the current production of fatty acid-based bioproducts in Y. lipolytica. To this end, we briefly provide the fatty acid metabolism, including fatty acid biosynthesis, transportation, and degradation. Then, we introduce the various metabolic engineering strategies for increasing bioproduct accumulation in Y. lipolytica. Further, the advanced progress in the production of fatty acid-based bioproducts by Y. lipolytica, including nutraceuticals, biofuels, and biochemicals, is summarized. This review will provide attractive thoughts for researchers working in the field of Y. lipolytica.

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De novo Biosynthesis of Odd-Chain Fatty Acids in Yarrowia lipolytica Enabled by Modular Pathway Engineering

Cited by 47 publications

References 59 publications

A common approach for absolute quantification of short chain CoA thioesters in prokaryotic and eukaryotic microbes

A common approach for absolute quantification of short chain CoA thioesters in prokaryotic and eukaryotic microbes

A common approach for absolute quantification of short chain CoA thioesters in industrially relevant gram-positive and gram-negative prokaryotic and eukaryotic microbes

Yarrowia lipolytica as an Oleaginous Platform for the Production of Value-Added Fatty Acid-Based Bioproducts

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