Study on the Synthesis of Polyunsaturated Fatty Acids by FattyAcid Synthase Pathway of <i>Schizochytrium</i> sp.

doi:10.13523/j.cb.20180904

China Biotechnology

2018, Vol. 38

Issue (9): 27-34 DOI: 10.13523/j.cb.20180904

Orginal Article

Study on the Synthesis of Polyunsaturated Fatty Acids by FattyAcid Synthase Pathway of Schizochytrium sp.

Shu-xia MA^1,³,Ling ZHANG²,Jin-fei YAN^1,^**(

),Song YOU³

¹ The Institute of Seawater Desalination and Comprehensive Utilization, Tianjin 300192, China
² Children’s Hospital, Capital Institute of Pediatrics,Beijing 100020,China;
³ Shenyang Pharmaceutical University, Shenyang 110016, China

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Abstract

As an excellent species for the production of DHA, Schizochytrium sp. has two distinct pathways for producing DHA, fatty acid synthase pathway and polyketide synthase pathway, of which fatty acid synthase pathway played limited role in the DHA synthesis process due to its low activity of one or more of the enzymes, resulted in its only responsible for some short chain fatty acid synthesis. In order to enhance the ability of fatty acid synthase pathway for producing DHA in Schizochytrium sp. to improve the DHA content of its oil, the exogenous △12-desaturase expression plasmid with screening markers for Schizochytrium sp.ATCC20888 from the brown algae was constructed, then Schizochytrium sp. were transformed through electroporation. After long-term screening, two positive transformants were obtained by DNA and RNA identification. Their initial biomass measurements were 11.14% and 4.12% higher than the wild strains while the DHA content in oils and fats were 19.50% and 14.65% higher than the wild strains.

Key words： Genetic engineering Fatty acid synthase pathway Schizochytrium sp. DHA

Received: 09 April 2018 Published: 12 October 2018

Corresponding Authors: Jin-fei YAN E-mail: jinfei_yan@qq.com

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	Shu-xia MA
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Cite this article:

Shu-xia MA,Ling ZHANG,Jin-fei YAN,Song YOU. Study on the Synthesis of Polyunsaturated Fatty Acids by FattyAcid Synthase Pathway of Schizochytrium sp.. China Biotechnology, 2018, 38(9): 27-34.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20180904 OR https://manu60.magtech.com.cn/biotech/Y2018/V38/I9/27

Fig.1 Expression vector of pBlu2SKP-Pht△12

Fig.2 Construction of plasmid pBlu2SKP-Pht△12-1UP: Fragment of Upoly-PET; NCR: Fragment of neo-CYC-RB18S; UPNCR: Fragment of Upoly-PET-neo-CYC-RB18S; QUPNCR: Fragment of Upoly-PET-neo-CYC-RB18S after digestion of Spe I, BamH I; PSKII: Plasmid of pBluescript II SK (+); QPSKII: Plasmid of pBluescript II SK (+) after digestion of Spe I, BamH I; M:DL5000

Fig.3 Construction of plasmid pBlu2SKP-Pht△12-2LU: Fragment of LB18S-Ubi; QLU: Fragment of LB18S-Ubi after digestion of Sac I, Not I; P-1: Plasmid of pBlu2SKP-Pht△12-1; QP-1: Plasmid of pBlu2SKP-Pht△12-1 after digestion of Sac I, Not I; M:DL5000

Fig.4 Construction of plasmid pBlu2SKP-Pht△12Qpht12:Fragment of LB18S-Ubi after digestion of Spe I, Not I; P-2: Plasmid of pBlu2SKP-Pht△12-2; QP-2: Plasmid of pBlu2SKP-Pht△12-1 after digestion of Spe I, Not I;P-3: Plasmid pBlu2SKP-Pht△12; QP-3: Plasmid pBlu2SKP-Pht△12 after digestion of Xmn I; M:DL5000

Fig.5 Experimental results of resistance screening

Fig.6 DNA level screening of transformantsThe positive bands on 1.2kb are fragments of functional gene pht12 amplified by primer Jh-F/R

Fig.7 RT-PCR results of the transformantsH: The fragment amplified by primer Jh-F/R, a part of the functional gene pht12

Table 1 Biomass comparison between the wild and thetransformants

Fig.8 Gas chromatograms of the transformants and the wild

Table 2 Percentage content of fatty acids in the oil of the transformants and the wild

Fig.9 Polyunsaturated fatty acidssynthesisof FAS pathway^[5]


[1]	曹万新, 孟橘, 田玉霞 . DHA的生理功能及应用研究进展. 中国油脂, 2011,36(3):1-4.

[1]	Cao W X, Meng J, Tian Y X . Research advance in application and physiological functions of DHA. China Oils and Fats, 2011,36(3):1-4.

[2]	陈殊贤, 郑晓辉 . 微藻油和鱼油中DHA的特性及应用研究进展. 食品科学, 2013,34(21):439-444.

[2]	Chen S X, Zheng X H . Research progress in characteristics and applications of DHA in microalga oil and fish oil. Food Science, 2013,34(21):439-444.

[3]	Ashford A, Barclay W R, Weaver C A , et al. Electron microscopy may reveal structure of docosahexaenoic acid-rich oil with in Schizochytrium sp. Lipids, 2000,35(12):1377-1386. doi: 10.1007/s11745-000-0655-2

[4]	Qiu X . Biosynthesis of docosahexaenoic acid (DHA, 22:6-4, 7,10,13,16,19): two distinct pathways. Prostaglandins Leukot Essent Fatty Acids, 2003,68(2):181-186. doi: 10.1016/S0952-3278(02)00268-5

[5]	Ratledge C . Fatty acid biosynthesis in microorganisms being used for single cell oil production. Biochimie, 2004,86(11):807-815. doi: 10.1016/j.biochi.2004.09.017

[6]	Hauvermale A, Kuner J, Rosenzweig B D , et al. Fatty acid production in Schizochytrium sp.: Involvement of a polyunsaturated fatty acid synthase and a type I fatty acid synthase. Lipids, 2006,41(8):739-747. doi: 10.1007/s11745-006-5025-6

[7]	Metz J G, Roessler P, Facciotti D , et al. Production of polyunsaturated fatty acids by polyketide synthases in both prokaryotes and eukaryotes. Science, 2001,293(5528):290-300. doi: 10.1126/science.1059593

[8]	Lippmeier J C, Crawford K S, Owen C B , et al. Characterization of both polyunsaturated fatty acid biosynthetic pathways in Schizochytrium sp. Lipids, 2009,44(7):621-630. doi: 10.1007/s11745-009-3311-9

[9]	Song X, Tan Y, Liu Y , et al. Different impacts of short-chain fatty acids on saturated and polyunsaturated fatty acid biosynthesis in Aurantiochytrium sp. SD116. Journal of Agricultural & Food Chemistry, 2013,61(41):9876-9881.

[10]	Nagano N, Sakaguchi K, Taoka Y , et al. Detection of genes involved in fatty acid elongation and desaturation in thraustochytrid marine eukaryotes. Journal of Oleo Science, 2011,60(9):475-481. doi: 10.5650/jos.60.475

[11]	Re L, Zhuang X, Chen S , et al. Introduction of ω-3 desaturase obviously changed the fatty acid profile and sterol content of Schizochytrium sp. Journal of Agricultural & Food Chemistry, 2015,63(44):9770-9776.

[12]	闫晋飞, 杨玉莹, 马淑霞 . 微藻基因工程概述. 生物学通报, 2017,52(4):1-5.

[12]	Yan J F, Yang Y Y, Ma S X . Overview of microalgae gene engineering. Bulletin of Biology, 2017,52(4):1-5.

[13]	Yan J, Cheng R, Lin X , et al. Overexpression of acetyl-CoA synthetase increased the biomass and fatty acid proportion in microalga Schizochytrium. Applied Microbiology & Biotechnology, 2013,97(5):1933-1939.

[14]	Bligh E G, Dyer W J . A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry & Physiology, 1959,37(8):911-917.

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