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Occurrence of long-chain fatty acids and glycolipids in the cell envelope fractions of baker's yeast.

ABSTRACT: 1. The total yield of fatty acids from the whole envelopes was markedly higher than that obtained from the ordinary cell walls. In both samples the major fatty acids were C(16) and C(18) acids. 2. The whole envelopes contained C(18) acids and long-chain (C(19)-C(26)) fatty acids, in a higher proportion than did the ordinary cell walls. Fifteen fatty acids with more than 18 carbon atoms were identified, among which 2-hydroxy-C(26:0) and C(26:0) acids predominated. 3. A complex sphingolipid containing inositol, phosphorus and mannose was isolated from the whole cell envelopes. The main fatty acids of this lipid were 2-hydroxy-C(26:0) and C(26:0) acids. It was concluded that this sphingolipid is present both in the ordinary cell wall and in the plasma membrane of baker's yeast. 4. The neutral lipids amounted to over 50% and the glycerophosphatides to about 30% of the total fatty acid content of the whole envelope. The major fatty acids in these lipids were C(16:1), C(18:1) and C(16:0) acids. The proportion of fatty acids with more than 18 carbon atoms was lowest in the neutral lipids, whereas the neutral glycolipids contained the highest percentage of these fatty acids. Acidic glycolipids amounted to 14% of the total fatty acid content of the whole envelope. The presence of a cerebroside sulphate in this lipid fraction was demonstrated, whereas the high content of 2-hydroxy-C(26:0) acid found is caused by the complex inositol- and mannose-containing sphingolipid.

SUBMITTER: Nurminen T

PROVIDER: S-EPMC1178257 | biostudies-other | 1971 Dec

REPOSITORIES: biostudies-other

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Project description:Concerns about global warming, fossil-fuel depletion, food security, and human health have promoted metabolic engineers to develop tools/strategies to overproduce microbial functional oils directly from renewable resources. Medium-chain fatty acids (MCFAs, C8-C12) have been shown to be important sources due to their diverse biotechnological importance, providing benefits ranging from functional lipids to uses in bio-fuel production. However, oleaginous microbes do not carry native pathways for the production of MCFAs, and therefore, diverse approaches have been adapted to compensate for the requirements of industrial demand. Mucor circinelloides is a promising organism for lipid production (15-36% cell dry weight; CDW) and the investigation of mechanisms of lipid accumulation; however, it mostly produces long-chain fatty acids (LCFAs). To address this challenge, we genetically modified strain M. circinelloides MU758, first by integrating heterologous acyl-ACP thioesterase (TE) into fatty acid synthase (FAS) complex and subsequently by modifying the ?-oxidation pathway by disrupting the acyl-CoA oxidase (ACOX) and/or acyl-CoA thioesterase (ACOT) genes with a preference for medium-chain acyl-CoAs, to elevate the yield of MCFAs. The resultant mutant strains (M-1, M-2, and M-3, respectively) showed a significant increase in lipid production in comparison to the wild-type strain (WT). MCFAs in M-1 (47.45%) was sharply increased compared to the wild type strain (2.25%), and it was further increased in M-2 (60.09%) suggesting a negative role of ACOX in MCFAs production. However, MCFAs in M-3 were much decreased compared to M-1,suggesting a positive role of ACOT in MCFAs production. The M-2 strain showed maximum lipid productivity (~1800 milligram per liter per day or mg/L.d) and MCFAs productivity (~1100 mg/L.d). Taken together, this study elaborates on how the combination of two multidimensional approaches, TE gene over-expression and modification of the ?-oxidation pathway via substantial knockout of specific ACOX gene, significantly increased the production of MCFAs. This synergistic approach ultimately offers a novel opportunity for synthetic/industrial biologists to increase the content of MCFAs.

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Occurrence of long-chain fatty acids and glycolipids in the cell envelope fractions of baker's yeast.

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