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Identification dehydratase domains from Schizochytrium sp. and Shewanella sp. and distinct functions in biosynthesis of fatty acids

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Abstract

Polyunsaturated fatty acid (PUFA) synthase is a special and effective enzyme for PUFA synthesis, and dehydratase (DH) domain played a crucial role in it. In this work, we compared four different DH domains from different strains (Schizochytrium sp. HX-308 and Shewanella sp. BR-2) and different gene clusters. First bioinformatics analysis showed that DH1, 2 and DH3 were similar to FabA and PKS-DH, respectively, and all of them got a hot-dog structure. Second, four DH domains were expressed in Escherichia coli that increased biomass. Especially, Schi-DH1,2 presented the highest dry cell weight of 2.3 g/L which was 1.62 times of that of control. Fatty acids profile analysis showed that DH1,2 could enhance the percentage of unsaturated fatty acids, especially DH1,2 from Schizochytrium sp., while DH3 benefited for the saturated fatty acid biosynthesis. Furthermore, five kinds of fatty acids were added to the medium to study the substrate preferences. Results revealed that DH1,2 domain preferred to acting on C16:0, while DH3 domain trended acting on C14:0 and C15:0, which illustrated DH from different clusters do have specific substrate preference. Besides, DH expression could save the cell growth inhibition by mid-chain fatty acids. This study provided more information about the catalysis mechanism of polyunsaturated fatty acid synthase and might promote the modification study based on this enzyme.

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References

  1. Rogers K, Valentine J, Keim A (2013) DHA supplementation: current implications in pregnancy and childhood. Pharmacol Res 70(1):13–19

    Article  CAS  Google Scholar 

  2. Chia-Ray L, Tsan-Mei C, Anin L et al (2019) Omega-3 polyunsaturated fatty acids suppress metastatic features of human cholangiocarcinoma cells by suppressing twist. Nutr Biochem 74:1–8

    Google Scholar 

  3. Ghasemi Fard S, Wang F, Sinclair J et al (2019) How does high DHA fish oil affect health? A systematic review of evidence. Crit Rev Food Sci Nutr 59:1684–1727

    Article  CAS  Google Scholar 

  4. Giltay J, Geleijnse JM, Kromhout D (2011) Effects of n-3 fatty acids on depressive symptoms and dispositional optimism after myocardial infarction. Am Soc Nutr 94:1442–1450

    CAS  Google Scholar 

  5. Hauvermale A, Kuner J, Rosenzwein B et al (2006) Fatty acid production in Schizochytrium sp.: involvement of a polyunsaturated fatty acid synthase and a type I fatty acid synthase. Lipids 41(8):739

    Article  CAS  Google Scholar 

  6. Lippmeier JC, Crawford KS, Owen CB et al (2009) Characterization of both polyunsaturated fatty acid biosynthetic pathways in Schizochytrium sp. Lipids 44(7):621–630

    Article  CAS  Google Scholar 

  7. Metz JG, Roessler P, Facciotti D et al (2001) Production of polyunsaturated fatty acids by polyketide synthases in both prokaryotes and eukaryotes. Science (New York, NY) 293(5528):290–293

    Article  CAS  Google Scholar 

  8. Naka I, Hayashi K et al (2019) Subtle control of carbon chain length in polyunsaturated fatty acid synthases. ACS Chem Biol 14(12):2553–2556

    Article  CAS  Google Scholar 

  9. Qu L, Xiaojun J, Lujing R et al (2011) Enhancement of docosahexaenoic acid production by Schizochytium sp. using a two-stage oxygen supply control strategy based on oxygen transfer coefficient. Lett Appl Microbiol 52(1):22–27

    Article  CAS  Google Scholar 

  10. Yazawa K (1996) Production of eicosapentaenoic acid from marine bacteria. Lipids 31:S297-300

    Article  CAS  Google Scholar 

  11. Hayashi N, Ikeuchi K et al (2019) Control mechanism for carbon-chain length in polyunsaturated fatty-acid synthases. Angew Chem Int Ed Engl 58(20):6605–6610

    Article  CAS  Google Scholar 

  12. La R (2012) Role of serine140 in the mode of action of mycobacterium tuberculosis β-ketoacyl-acp reductase (maba). BMC Res Notes 5(1):1–10

    Article  Google Scholar 

  13. Napier JA, Michaelson LV (2001) Genomic and functional characterization of polyunsaturated fatty acid biosynthesis in caenorhabditis elegans. Lipids 36(8):761–766

    Article  CAS  Google Scholar 

  14. Hayashi S, Dairi T (2020) Off-loading mechanism of products in polyunsaturated fatty acid synthases. ACS Chem Biol 15(3):651–656

    Article  CAS  Google Scholar 

  15. Jiang H, Zirkle R, Metz JG et al (2008) The role of tandem acyl carrier protein domains in polyunsaturated fatty acid biosynthesis. Am Chem Soc 130(20):6336

    Article  CAS  Google Scholar 

  16. Trujillo U, Vazquez-Rosa E, Oyola-Robles D et al (2013) Solution structure of the tandem acyl carrier protein domains from a polyunsaturated fatty acid synthase reveals beads-on-a-string configuration. PLoS ONE 8(2):e57859

    Article  CAS  Google Scholar 

  17. Almendariz-Palacios C, Meesapyodsuk D, Qiu X (2021) Functional analysis of an acyltransferase-like domain from polyunsaturated fatty acid synthase in Thraustochytrium. Microorganisms 9(3):626

    Article  CAS  Google Scholar 

  18. Lujing R, Shenglan C, Lingjun G et al (2018) Exploring the function of acyltransferase and domain replacement in order to change the polyunsaturated fatty acid profile of Schizochytrium sp. Algal Res 29:193–201

    Article  Google Scholar 

  19. Donadio S, Mcalpine JB, Shelden PJ et al (1993) An erythromycin analog produced by reprogramming of polyketide synthesis. Proc Natl Acad Sci USA 90(15):7119–7123

    Article  CAS  Google Scholar 

  20. Zhang M, Zhang H, Li Q et al (2021) Structural insights into the trans-acting enoyl reductase in the biosynthesis of long-chain polyunsaturated fatty acids in Shewanella piezotolerans. J Agric Food Chem 69(7):2316–2324

    Article  CAS  Google Scholar 

  21. Liu Z, Zang X, Cao X et al (2018) Cloning of the pks3 gene of aurantiochytrium limacinum and functional study of the 3-ketoacyl-acp reductase and dehydratase enzyme domains. PLoS ONE 13(12):e02080533

    Google Scholar 

  22. Liu L, Hu Z, Li S et al (2020) Comparative transcriptomic analysis uncovers genes responsible for the DHA enhancement in the mutant Aurantiochytrium sp. Microorganisms 8(4):529

    Article  CAS  Google Scholar 

  23. Shi Y, Chen Z, Li Y et al (2021) Function of orfc of the polyketide synthase gene cluster on fatty acid accumulation in Schizochytrium limacinum SR21. Biotechnol Biofuels 14(1):1–14

    Article  Google Scholar 

  24. Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37(8):911–917

    Article  CAS  Google Scholar 

  25. Xiaoyan Z, Shenglan C, Lujing R et al (2015) Construction and verification of genetic transformation system of Schizochytrium based on homologous recombination. Acta Microbiol Sin 55(4):510–517

    Google Scholar 

  26. Ma L, Pa LR (2007) Clustal w and clustal x version 2.0. Bioinformatics 2:2947–2948

    Google Scholar 

  27. Xavier R, Patrice G (2014) Deciphering key features in protein structures with the new endscript server. Nucleic Acids Res 42(W1):W320–W324

    Article  Google Scholar 

  28. Kelley LA (2015) The phyre2 web portal for protein modeling, prediction and analysis. Nat Protoc 10:845–858

    Article  CAS  Google Scholar 

  29. Oyola-Robles G, Trujillo D et al (2013) Identification of novel protein domains required for the expression of an active dehydratase fragment from a polyunsaturated fatty acid synthase. Protein Sci 22(7):954–963

    Article  CAS  Google Scholar 

  30. Qiu X, Xie X, Meesapyodsuk D et al (2020) Molecular mechanisms for biosynthesis and assembly of nutritionally important very long chain polyunsaturated fatty acids in microorganisms. Prog Lipid Res 79:101047

    Article  CAS  Google Scholar 

  31. Liu W, Schmidt B, Muller-Wittig W (2011) Cuda-blastp: accelerating blastp on cuda-enabled graphics hardware. IEEE ACM Trans Comput Biol Bioinf 8:1678–1684

    Article  Google Scholar 

  32. Brock H, Kass LR, Bloch K (1967) B-hydroxydecanoyl thioester dehydrase. J Biol Chem 242:4432–4440

    Article  CAS  Google Scholar 

  33. Yoshida K, Hashimoto M, Hori R et al (2016) Bacterial long-chain polyunsaturated fatty acids: their biosynthetic genes, functions, and practical use. Mar Drugs 14(5):94

    Article  Google Scholar 

  34. Dominik AH, Roman PJ, Zähringer F et al (2016) Mycocerosic acid synthase exemplifies the architecture of reducing polyketide synthases. Nature 531:533–537

    Article  Google Scholar 

  35. Xie X, Meesapyodsuk D, Qiu X (2017) Functional analysis of the dehydratase domains of a PUFA synthase from thraustochytrium in Escherichia coli. Appl Environ Microb 102(2):847–856

    Google Scholar 

  36. Oyola-Robles D, Rullan-Lind C, Carballeira N et al (2014) Expression of dehydratase domains from a polyunsaturated fatty acid synthase increases the production of fatty acids in Escherichia coli. Enzyme Microb Technol 55:133–139

    Article  CAS  Google Scholar 

  37. Liu P, Chernyshov A, Najdi T et al (2013) Membrane stress caused by octanoic acid in Saccharomyces cerevisiae. Appl Microbiol Biotechnol 97(7):3239–3251

    Article  CAS  Google Scholar 

  38. Borrull A, Lopez-Martinez G, Poblet M et al (2015) New insights into the toxicity mechanism of octanoic and decanoic acids on Saccharomyces cerevisiae. Yeast 32(5):451–460

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was financially supported by the National Key Research and Development Program of China (No. 2019YFA0905700), the National Natural Science Foundation of China (No. 21878151) and the Natural Science Foundation of Jiangsu Province (BK20211535).

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Authors and Affiliations

  1. College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No. 30 South Puzhu Road, Nanjing, 211816, People’s Republic of China

    Yanli Man, Yuting Zhang, Jiayi Jiang, Quanyu Zhao & Lujing Ren

  2. School of Pharmaceutical Sciences, Nanjing Tech University, No. 30 South Puzhu Road, Nanjing, 211816, People’s Republic of China

    Yanli Man, Yuting Zhang, Jiayi Jiang, Quanyu Zhao & Lujing Ren

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  1. Yanli Man
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  2. Yuting Zhang
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Correspondence to Lujing Ren.

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Man, Y., Zhang, Y., Jiang, J. et al. Identification dehydratase domains from Schizochytrium sp. and Shewanella sp. and distinct functions in biosynthesis of fatty acids. Bioprocess Biosyst Eng 45, 107–115 (2022). https://doi.org/10.1007/s00449-021-02644-1

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