Identification and characterization of fatty acid desaturases in Schizochytrium sp. HX-308

Jia, Yu-Lei; Wang, Yu‐Zhou; Nong, Fang-Tong; Ma, Wang; Huang, Pengwei; Sun, Xiao‐Man

doi:10.1016/j.algal.2022.102861

Cited by 4 publications

(7 citation statements)

References 52 publications

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“…The plasmid contains the gene encoding URA3, a nutrient-deficient gene that has been used extensively in the screening of Y. lipolytica strains . As shown in Figure S1, all engineered strains have grown single clones on URA3-deficient plates.…”

Section: Resultsmentioning

confidence: 99%

“…The reagents and methods used for plasmid construction and transformation, as well as the medium and conditions required for the fermentation of Y. lipolytica in this work, were the same as in our previous reports. 24 2.3. DCW, Total Lipid, and Fatty Acid Analysis.…”

Section: Cloning and Bioinformatics Analysis Of Genesmentioning

confidence: 99%

“…The methods of DCW determination, lipid extraction, and fatty acid methyl esters (FAMEs) were the same as in our previous reports. 24 The prepared FAME samples were analyzed using a GC-2030 gas chromatography system (GC-2030, Shimadzu, Japan).…”

Section: Cloning and Bioinformatics Analysis Of Genesmentioning

confidence: 99%

“…The plasmid contains the gene encoding URA3, a nutrient-deficient gene that has been used extensively in the screening of Y. lipolytica strains. 24 As shown in Figure S1, all engineered strains have grown single clones on URA3-deficient plates. Subsequently, the genomic DNA and RNA of Y. lipolytica were extracted as templates for PCR and qPCR, respectively.…”

Section: Heterologous Expression Of Orfa Orfb and Orfc In Y Lipolytic...mentioning

confidence: 99%

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Function of the Polyketide Synthase Domains of Schizochytrium sp. on Fatty Acid Synthesis in Yarrowia lipolytica

Jia

Nong

et al. 2023

J. Agric. Food Chem.

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It is well known that polyunsaturated fatty acids (PUFAs) in Schizochytrium sp. are mainly synthesized via the polyketide synthase (PKS) pathway. However, the specific mechanism of PKS in fatty acid synthesis is still unclear. In this work, the functions of ORFA, ORFB, ORFC, and their individual functional domain genes on fatty acid synthesis were investigated through heterologous expression in Yarrowia lipolytica. The results showed that the expression of ORFA, ORFB, ORFC, and their individual functional domains all led to the increase of the very long-chain PUFA content (mainly eicosapentaenoic acid). Furthermore, the transcriptomic analysis showed that except for the 3-ketoacyl-ACP synthase (KS) domain of ORFB, the expression of an individual functional domain, including malonyl-CoA: ACP acyltransferase, 3-hydroxyacyl-ACP dehydratase (DH), 3-ketoacyl-ACP reductase, and KS domains of ORFA, acyltransferase domains of ORFB, and two DH domains of ORFC resulted in upregulation of the tricarboxylic acid cycle and pentose phosphate pathway, downregulation of the triacylglycerol biosynthesis, fatty acid synthesis pathway, and β-oxidation in Yarrowia lipolytica. These results provide a theoretical basis for revealing the function of PKS in fatty acid synthesis in Y. lipolytica and elucidate the possible mechanism for PUFA biosynthesis.

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Section: Resultsmentioning

confidence: 99%

Section: Cloning and Bioinformatics Analysis Of Genesmentioning

confidence: 99%

Section: Cloning and Bioinformatics Analysis Of Genesmentioning

confidence: 99%

Section: Heterologous Expression Of Orfa Orfb and Orfc In Y Lipolytic...mentioning

confidence: 99%

See 2 more Smart Citations

Function of the Polyketide Synthase Domains of Schizochytrium sp. on Fatty Acid Synthesis in Yarrowia lipolytica

Jia

Nong

et al. 2023

J. Agric. Food Chem.

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“…The URA3 maker was recycled by selection on YPD plates supplemented with 1 g/L 5-fluoroorotic acid (5-FOA) and 15 g/L agar. The medium and conditions required for the fermentation of Y. lipolytica in this work were also described previously [ 45 ].…”

Section: Methodsmentioning

confidence: 99%

Identification of lipid synthesis genes in Schizochytrium sp. and their application in improving eicosapentaenoic acid synthesis in Yarrowia lipolytica

Jia,

Zhang,

et al. 2024

Biotechnol Biofuels

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Background Eicosapentaenoic acid (EPA) is widely used in the functional food and nutraceutical industries due to its important benefits to human health. Oleaginous microorganisms are considered a promising alternative resource for the production of EPA lipids. However, the storage of EPA in triglyceride (TG) becomes a key factor limiting its level. Results This study aimed to incorporate more EPA into TG storage through metabolic engineering. Firstly, key enzymes for TG synthesis, the diacylglycerol acyltransferase (DGAT) and glycerol-3-phosphate acyltransferase (GPAT) genes from Schizochytrium sp. HX-308 were expressed in Yarrowia lipolytica to enhance lipid and EPA accumulation. In addition, engineering the enzyme activity of DGATs through protein engineering was found to be effective in enhancing lipid synthesis by replacing the conserved motifs “HFS” in ScDGAT2A and “FFG” in ScDGAT2B with the motif “YFP”. Notably, combined with lipidomic analysis, the expression of ScDGAT2C and GPAT2 enhanced the storage of EPA in TG. Finally, the accumulation of lipid and EPA was further promoted by identifying and continuing to introduce the ScACC, ScACS, ScPDC, and ScG6PD genes from Schizochytrium sp., and the lipid and EPA titer of the final engineered strain reached 2.25 ± 0.03 g/L and 266.44 ± 5.74 mg/L, respectively, which increased by 174.39% (0.82 ± 0.02 g/L) and 282.27% (69.70 ± 0.80 mg/L) compared to the initial strain, respectively. Conclusion This study shows that the expression of lipid synthesis genes from Schizochytrium sp. in Y. lipolytica effectively improves the synthesis of lipids and EPA, which provided a promising target for EPA-enriched microbial oil production.

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Deciphering and regulating carotenoid synthesis using norflurazon to enhance fatty acid synthesis in oleaginous marine protist Schizochytrium sp. S31

Guo

Zeng

et al. 2023

Industrial Crops and Products

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Identification and characterization of fatty acid desaturases in Schizochytrium sp. HX-308

Cited by 4 publications

References 52 publications

Function of the Polyketide Synthase Domains of Schizochytrium sp. on Fatty Acid Synthesis in Yarrowia lipolytica

Function of the Polyketide Synthase Domains of Schizochytrium sp. on Fatty Acid Synthesis in Yarrowia lipolytica

Identification of lipid synthesis genes in Schizochytrium sp. and their application in improving eicosapentaenoic acid synthesis in Yarrowia lipolytica

Deciphering and regulating carotenoid synthesis using norflurazon to enhance fatty acid synthesis in oleaginous marine protist Schizochytrium sp. S31

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