Fatty acid ethyl esters production in aqueous phase by the oleaginous yeast Rhodosporidium toruloides

Jin, Guojie; Zhang, Yixin; Shen, Hongwei; Yang, Xiaobing; Xie, Haibo; Zhao, Zongbao K.

doi:10.1016/j.biortech.2013.10.023

Cited by 27 publications

(20 citation statements)

References 31 publications

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“…The results showed that RtDGATa did not have DGAT activity and did not enhance the sterol ester biosynthesis capacity of the complemented S. cervisiae HY4, although RtDGATa belonged to DGAT1 family and shared substantial similarity with ScARE. Furthermore the level of sterol ester in R. toruloides wild type was extremely low (Jin et al, 2013) and these results were consistent with mRNA expression level of RtDGATa in R. toruloides.…”

Section: Discussionsupporting

confidence: 75%

“…The lipid content in red yeast R. toruloides was up to 70% of its biomass under certain conditions and total lipids mainly composed with neutral lipids (mainly TAG and DAG) (Jin et al, 2013). DGAT, which catalyzes TAG formation from DAG and fatty acyl-CoA, is the terminal step for lipid accumulation.…”

Section: Discussionmentioning

confidence: 99%

“…It can accumulate lipids up to 70% of its biomass under certain conditions and the content of neutral lipids (mainly TAG and DAG) was up to over 90% of total lipids (Jin et al, 2013). However, the molecular mechanism of TAG biosynthesis in R. toruloides had barely been studies.…”

Section: Introductionmentioning

confidence: 99%

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Identification and Characterization of Diacylglycerol Acyltransferase in Oleaginous Yeast <i>Rhodosporidium toruloides</i>

Wang¹,

Zhang²,

Zhao³

et al. 2016

American Journal of Biochemistry and Biotechnology

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Diacylglycerol acyltransferase (DGAT), which catalyzes TAG formation from DAG and acyl-CoA, has been considered to play a vital role in TAG accumulation in oleaginous microorganisms. The genome of oleaginous yeast Rhodosporidium toruloides contains two putative DGAT genes, RtDGATa and RtDGATb, which shared little conserved amino acid coding sequence with each other. Phylogeny tree analysis showed that RtDGATa belonged to DGAT1 family and RtDGATb belonged to DGAT2 family. For functional identification of the DGATs, RtDGATa and RtDGATb were individually expressed in Saccharomyces cerevisiae TAGdeficient quadruple mutant (H1246). RtDGATa had obvious preference for monounsaturated fatty acids, however the expression of RtDGATa did not alter the TAG content in S. cerevisiae H1246 and it had non-involvement in TAG accumulation according to its mRNA expression level in R. toruloides. The expression of RtDGATb could completely resume TAG biosynthesis in S. cerevisiae H1246. Substrate preference experiments revealed that RtDGATb preferred unsaturated fatty acids over saturated fatty acids, but not C18:3. Only the expression pattern of RtDGATb was related to the process of fatty acid biosynthesis, suggesting that RtDGATb plays an important role in lipid accumulation in R. toruloides.

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Section: Discussionsupporting

confidence: 75%

Section: Discussionmentioning

confidence: 99%

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Identification and Characterization of Diacylglycerol Acyltransferase in Oleaginous Yeast <i>Rhodosporidium toruloides</i>

Wang¹,

Zhang²,

Zhao³

et al. 2016

American Journal of Biochemistry and Biotechnology

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“…Recently, many efforts have been undertaken to engineer S. cerevisiae or Escherichia coli for the production of lipid-derived metabolites and biofuels, but the efficiency remained fairly low (14,57). Alternatively, it has been shown that more than 70% of the TAGs in R. toruloides can be converted to fatty acid ethyl esters when incubating lipid-rich cells with 10% ethanol in an aqueous environment (58). It is believed that the LD provides the hydrophobic microenvironment for LD-associated lipases, which traditionally catalyzed the transesterification in nonaqueous media (59).…”

Section: Discussionmentioning

confidence: 99%

Dynamics of the Lipid Droplet Proteome of the Oleaginous Yeast Rhodosporidium toruloides

et al. 2015

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c Lipid droplets (LDs) are ubiquitous organelles that serve as a neutral lipid reservoir and a hub for lipid metabolism. Manipulating LD formation, evolution, and mobilization in oleaginous species may lead to the production of fatty acid-derived biofuels and chemicals. However, key factors regulating LD dynamics remain poorly characterized. Here we purified the LDs and identified LD-associated proteins from cells of the lipid-producing yeast Rhodosporidium toruloides cultured under nutrient-rich, nitrogen-limited, and phosphorus-limited conditions. The LD proteome consisted of 226 proteins, many of which are involved in lipid metabolism and LD formation and evolution. Further analysis of our previous comparative transcriptome and proteome data sets indicated that the transcription level of 85 genes and protein abundance of 77 proteins changed under nutrient-limited conditions. Such changes were highly relevant to lipid accumulation and partially confirmed by reverse transcription-quantitative PCR. We demonstrated that the major LD structure protein Ldp1 is an LD marker protein being upregulated in lipid-rich cells. When overexpressed in Saccharomyces cerevisiae, Ldp1 localized on the LD surface and facilitated giant LD formation, suggesting that Ldp1 plays an important role in controlling LD dynamics. Our results significantly advance the understanding of the molecular basis of lipid overproduction and storage in oleaginous yeasts and will be valuable for the development of superior lipid producers. Lipid droplets (LDs), intracellular organelles with deposits of neutral lipids and involved in many cellular activities, are widely present in both eukaryotic and prokaryotic cells (1-4). These organelles consist of a neutral lipid core surrounded by a phospholipid monolayer and associated proteins (3, 5). It has been known that LDs serve as the energy reservoir of cells, which may increase the adaptation by mobilization and degradation of lipids during nutrient deprivation, and also connect with other cellular processes, including lipid transport, membrane biogenesis, lipotoxicity relief, protein storage and degradation, pathogenicity, and autophagy (6-9). Because the biology of LDs is closely linked to some diseases, such as obesity, type 2 diabetes, and atherosclerosis, great progress has been made in elucidating the cellular trafficking, dynamics, and biogenesis of LDs in mammalian cells (2, 7, 10). However, there have been few studies on LDs in other species, especially naturally lipid-producing microorganisms (11-13). Analysis of these microorganisms is motivated by the fact that microbial lipid production holds a great promise to convert waste materials, including lignocellulosic biomass, into fatty acid-derived fuel molecules and chemicals in a scenario of biorefinery and sustainable development (14, 15).The major components of LDs are neutral lipids, including triacylglycerols (TAGs), sterol esters, and ether lipids (16). Neutral lipids constitute more than 90% of LDs by weight, but the ratio of TAGs to ste...

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“…To synthesize FFAs directly from glucose, many research groups have attempted to develop and engineer yeast (Jin et al 2013) and Escherichia coli strains as production host cells (Liu et al 2010;Lu et al 2008). According to these works, releasing fatty acyl groups from acyl carrier protein (ACP) through amplification of thioesterase (TE), or the disruption of the FFA β-oxidation pathway by the deletion of acyl-CoA synthase (fadD), functioning as the first-step enzyme of the β-oxidation pathway, is critical for efficient synthesis of FFAs (Lu et al 2008;Jeon 2012).…”

Section: Introductionmentioning

confidence: 99%

The production of ω-hydroxy palmitic acid using fatty acid metabolism and cofactor optimization in Escherichia coli

Sung

Jung

Choi

et al. 2015

Appl Microbiol Biotechnol

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Hydroxylated fatty acids (HFAs) are used as important precursors for bulk and fine chemicals in the chemical industry. Here, to overproduce long-chain (C16-C18) fatty acids and hydroxy fatty acid, their biosynthetic pathways including thioesterase (Lreu_0335) from Lactobacillus reuteri DSM20016, β-hydroxyacyl-ACP dehydratase (fabZ) from Escherichia coli, and a P450 system (i.e., CYP153A from Marinobacter aquaeolei VT8 and camA/camB from Pseudomonas putida ATCC17453) were overexpressed. Acyl-CoA synthase (fadD) involved in fatty acid degradation by β-oxidation was also deleted in E. coli BW25113. The engineered E. coli FFA4 strain without the P450 system could produce 503.0 mg/l of palmitic (C16) and 508.4 mg/l of stearic (C18) acids, of which the amounts are ca. 1.6- and 2.3-fold higher than those of the wild type. On the other hand, the E. coli HFA4 strain including the P450 system for ω-hydroxylation could produce 211.7 mg/l of ω-hydroxy palmitic acid, which was 42.1 ± 0.1 % of the generated palmitic acid, indicating that the hydroxylation reaction was the rate-determining step for the HFA production. For the maximum production of ω-hydroxy palmitic acid, NADH, i.e., an essential cofactor for P450 reaction, was overproduced by the integration of NAD(+)-dependent formate dehydrogenase (FDH) from Candida boidinii into E. coli chromosome and the deletion of alcohol dehydrogenase (ADH). Finally, the NADH-level-optimized E. coli strain produced 610 mg/l of ω-hydroxy palmitic acid (ω-HPA), which was almost a threefold increase in its yield compared to the same strain without NADH overproduction.

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Fatty acid ethyl esters production in aqueous phase by the oleaginous yeast Rhodosporidium toruloides

Cited by 27 publications

References 31 publications

Identification and Characterization of Diacylglycerol Acyltransferase in Oleaginous Yeast <i>Rhodosporidium toruloides</i>

Identification and Characterization of Diacylglycerol Acyltransferase in Oleaginous Yeast <i>Rhodosporidium toruloides</i>

Dynamics of the Lipid Droplet Proteome of the Oleaginous Yeast Rhodosporidium toruloides

The production of ω-hydroxy palmitic acid using fatty acid metabolism and cofactor optimization in Escherichia coli

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