Functions of PKS Genes in Lipid Synthesis of Schizochytrium Sp. by Gene Disruption and Metabolomics Analysis

Overview

Journal Mar Biotechnol (NY)

Specialties Biology
Biotechnology

Date 2018 Aug 24

PMID 30136198

Citations 11

Authors

Zhipeng Li

Xi Chen

Jun Li

Tong Meng

Lingwei Wang

Zhen Chen

Yanyan Shi

Xueping Ling

Weiang Luo

Dafeng Liang

Yinghua Lu

Qingbiao Li

Ning He

Affiliations

Soon will be listed here.

Abstract

Schizochytrium sp. is a kind of marine microalgae with great potential as promising sustainable source of polyunsaturated fatty acids (PUFAs). Polyketide synthase-like (PKS synthase) is supposed to be one of the main ways to synthesize PUFAs in Schizochytrium sp. In order to study the exact relationship between PKS and PUFA biosynthesis, chain length factor (CLF) and dehydrogenase (DH) were cloned from the PKS gene cluster in Schizochytrium sp., then disrupted by homologous recombination. The results showed that DH- and CLF-disrupted strains had significant decreases (65.85 and 84.24%) in PUFA yield, while the saturated fatty acid (SFA) proportion in lipids was slightly increased. Meanwhile, the disruption of CLF decreased the C-22 PUFA proportion by 57.51% without effect on C-20 PUFA accumulation while DH-disrupted mutant decreased the production of each PUFA. Combined with analysis of protein prediction, it indicated that CLF gene exerted an enormous function on the carbon chain elongation in PUFA synthesis, especially for the final elongation from C-20 to C-22 PUFAs. Metabolomics analysis also suggested that the disruption of both genes resulted in the decrease of PUFAs but increase of SFAs, thus weakening glycolysis and tricarboxylic acid (TCA) cycle pathways. This study offers a broad new vision to research the mechanism of PUFA synthesis in Schizochytrium sp.

Citing Articles

Comparative Transcriptome Analysis Unveils Regulatory Factors Influencing Fatty Liver Development in Lion-Head Geese under High-Intake Feeding Compared to Normal Feeding.

Kong J, Yao Z, Chen J, Zhao Q, Li T, Dong M Vet Sci. 2024; 11(8).

PMID: 39195820 PMC: 11359645. DOI: 10.3390/vetsci11080366.

Selectively superior production of docosahexaenoic acid in Schizochytrium sp. through engineering the fatty acid biosynthetic pathways.

Liu Y, Han X, Chen Z, Yan Y, Chen Z Biotechnol Biofuels Bioprod. 2024; 17(1):75.

PMID: 38831337 PMC: 11145866. DOI: 10.1186/s13068-024-02524-2.

Engineering Fatty Acid Biosynthesis in Microalgae: Recent Progress and Perspectives.

Song Y, Wang F, Chen L, Zhang W Mar Drugs. 2024; 22(5).

PMID: 38786607 PMC: 11122798. DOI: 10.3390/md22050216.

Knockout of a PLD gene in Schizochytrium limacinum SR21 enhances docosahexaenoic acid accumulation by modulation of the phospholipid profile.

Zhang Y, Cui X, Lin S, Lu T, Li H, Lu Y Biotechnol Biofuels Bioprod. 2024; 17(1):16.

PMID: 38291531 PMC: 10826259. DOI: 10.1186/s13068-024-02465-w.

Biotechnological production of omega-3 fatty acids: current status and future perspectives.

Qin J, Kurt E, LBassi T, Sa L, Xie D Front Microbiol. 2023; 14:1280296.

PMID: 38029217 PMC: 10662050. DOI: 10.3389/fmicb.2023.1280296.

References

Wallis J, Watts J, Browse J . Polyunsaturated fatty acid synthesis: what will they think of next?. Trends Biochem Sci. 2002; 27(9):467. DOI: 10.1016/s0968-0004(02)02168-0. View

Sakaguchi K, Matsuda T, Kobayashi T, Ohara J, Hamaguchi R, Abe E . Versatile transformation system that is applicable to both multiple transgene expression and gene targeting for Thraustochytrids. Appl Environ Microbiol. 2012; 78(9):3193-202. PMC: 3346472. DOI: 10.1128/AEM.07129-11. View

Ratledge C . Fatty acid biosynthesis in microorganisms being used for Single Cell Oil production. Biochimie. 2004; 86(11):807-15. DOI: 10.1016/j.biochi.2004.09.017. View

Lee Chang K, Nichols C, Blackburn S, Dunstan G, Koutoulis A, Nichols P . Comparison of Thraustochytrids Aurantiochytrium sp., Schizochytrium sp., Thraustochytrium sp., and Ulkenia sp. for production of biodiesel, long-chain omega-3 oils, and exopolysaccharide. Mar Biotechnol (NY). 2014; 16(4):396-411. DOI: 10.1007/s10126-014-9560-5. View

Tang Y, Tsai S, Khosla C . Polyketide chain length control by chain length factor. J Am Chem Soc. 2003; 125(42):12708-9. DOI: 10.1021/ja0378759. View

Song X, Tan Y, Liu Y, Zhang J, Liu G, Feng Y . Different impacts of short-chain fatty acids on saturated and polyunsaturated fatty acid biosynthesis in Aurantiochytrium sp. SD116. J Agric Food Chem. 2013; 61(41):9876-81. DOI: 10.1021/jf403153p. View

Raghukumar S . Thraustochytrid Marine Protists: production of PUFAs and Other Emerging Technologies. Mar Biotechnol (NY). 2008; 10(6):631-40. DOI: 10.1007/s10126-008-9135-4. View

Ren L, Huang H, Xiao A, Lian M, Jin L, Ji X . Enhanced docosahexaenoic acid production by reinforcing acetyl-CoA and NADPH supply in Schizochytrium sp. HX-308. Bioprocess Biosyst Eng. 2009; 32(6):837-43. DOI: 10.1007/s00449-009-0310-4. View

Okuyama H, Orikasa Y, Nishida T, Watanabe K, Morita N . Bacterial genes responsible for the biosynthesis of eicosapentaenoic and docosahexaenoic acids and their heterologous expression. Appl Environ Microbiol. 2006; 73(3):665-70. PMC: 1800774. DOI: 10.1128/AEM.02270-06. View

10.

Huang J, Jiang X, Zhang X, Chen W, Tian B, Shu Z . Expressed sequence tag analysis of marine fungus Schizochytrium producing docosahexaenoic acid. J Biotechnol. 2008; 138(1-2):9-16. DOI: 10.1016/j.jbiotec.2008.07.1994. View

11.

Ling X, Guo J, Liu X, Zhang X, Wang N, Lu Y . Impact of carbon and nitrogen feeding strategy on high production of biomass and docosahexaenoic acid (DHA) by Schizochytrium sp. LU310. Bioresour Technol. 2014; 184:139-147. DOI: 10.1016/j.biortech.2014.09.130. View

12.

Metz J, Roessler P, Facciotti D, Levering C, Dittrich F, Lassner M . Production of polyunsaturated fatty acids by polyketide synthases in both prokaryotes and eukaryotes. Science. 2001; 293(5528):290-3. DOI: 10.1126/science.1059593. View

13.

Keatinge-Clay A, Maltby D, Medzihradszky K, Khosla C, Stroud R . An antibiotic factory caught in action. Nat Struct Mol Biol. 2004; 11(9):888-93. DOI: 10.1038/nsmb808. View

14.

Liu B, Liu J, Sun P, Ma X, Jiang Y, Chen F . Sesamol Enhances Cell Growth and the Biosynthesis and Accumulation of Docosahexaenoic Acid in the Microalga Crypthecodinium cohnii. J Agric Food Chem. 2015; 63(23):5640-5. DOI: 10.1021/acs.jafc.5b01441. View

15.

Uauy R, Hoffman D, Peirano P, Birch D, Birch E . Essential fatty acids in visual and brain development. Lipids. 2001; 36(9):885-95. DOI: 10.1007/s11745-001-0798-1. View

16.

Warude D, Joshi K, Harsulkar A . Polyunsaturated fatty acids: biotechnology. Crit Rev Biotechnol. 2006; 26(2):83-93. DOI: 10.1080/07388550600697479. View

17.

Matsuda T, Sakaguchi K, Hamaguchi R, Kobayashi T, Abe E, Hama Y . Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304. J Lipid Res. 2012; 53(6):1210-22. PMC: 3351828. DOI: 10.1194/jlr.M024935. View

18.

Yeh K, Chang J . Effects of cultivation conditions and media composition on cell growth and lipid productivity of indigenous microalga Chlorella vulgaris ESP-31. Bioresour Technol. 2011; 105:120-7. DOI: 10.1016/j.biortech.2011.11.103. View

19.

Zhang A, Ji X, Wu W, Ren L, Yu Y, Huang H . Lipid Fraction and Intracellular Metabolite Analysis Reveal the Mechanism of Arachidonic Acid-Rich Oil Accumulation in the Aging Process of Mortierella alpina. J Agric Food Chem. 2015; 63(44):9812-9. DOI: 10.1021/acs.jafc.5b04521. View

20.

Miao L, Wang Y, Chi S, Yan J, Guan G, Hui B . Reduction of fatty acid flux results in enhancement of astaxanthin synthesis in a mutant strain of Phaffia rhodozyma. J Ind Microbiol Biotechnol. 2010; 37(6):595-602. DOI: 10.1007/s10295-010-0706-9. View