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维生素A-DHA酯的酶法合成及纯化制备

高坤鹏 孙建安 毛相朝

高坤鹏, 孙建安, 毛相朝. 维生素A-DHA酯的酶法合成及纯化制备[J]. 南方水产科学, 2022, 18(2): 23-30. doi: 10.12131/20210288
引用本文: 高坤鹏, 孙建安, 毛相朝. 维生素A-DHA酯的酶法合成及纯化制备[J]. 南方水产科学, 2022, 18(2): 23-30. doi: 10.12131/20210288
GAO Kunpeng, SUN Jian'an, MAO Xiangzhao. Enzymatic synthesis and purification of Vitamin A docosahexaenoate[J]. South China Fisheries Science, 2022, 18(2): 23-30. doi: 10.12131/20210288
Citation: GAO Kunpeng, SUN Jian'an, MAO Xiangzhao. Enzymatic synthesis and purification of Vitamin A docosahexaenoate[J]. South China Fisheries Science, 2022, 18(2): 23-30. doi: 10.12131/20210288

维生素A-DHA酯的酶法合成及纯化制备

doi: 10.12131/20210288
基金项目: 国家重点研发计划项目 (2019YFD0901902)
详细信息
    作者简介:

    高坤鹏 (1996—),男,博士研究生,研究方向为油脂的酶法改性。E-mail: gaokunpeng@stu.ouc.edu.cn

    通讯作者:

    毛相朝 (1981—),男,教授,博士,从事食品酶学与生物催化相关研究。E-mail: xzhmao@ouc.edu.cn

  • 中图分类号: Q 814.9

Enzymatic synthesis and purification of Vitamin A docosahexaenoate

  • 摘要: 维生素A具有维持正常视力、改善皮肤、抗衰老等多种生理活性,但极不稳定,很容易在光、热、氧的作用下发生分解。选择二十二碳六烯酸乙酯 (Ethyl docosahexaenoate, EDHE) 与维生素A醋酸酯 (Vitamin A acetate, VAAE) 作为底物,在脂肪酶Novozyme435的催化下发生转酯反应,从而合成一种新型的维生素A-DHA酯 (Vitamin A docosahexaenoate, VADHE)。利用液相及质谱对目的产物进行鉴定,在此基础上,利用硅胶柱层析 (C18填料) 对反应体系中的VADHE进行纯化。经液相色谱检测,VADHE的纯度达到90%以上。为了进一步提高目的化合物得率,分别在有机溶剂体系和无溶剂体系中对转酯反应进行了优化。在有机溶剂体系中,反应6 h转化率可达40.61%。在无溶剂体系中,用100 mg脂肪酶催化反应4 h,转化率可达56.39%。该研究首次设计并合成了新型脂质VADHE,实现了该物质的纯化制备,为后期的活性检测实验奠定了基础。
  • 图  1  维生素A-DHA酯 (VADHE) 的液相 (a) 和质谱 (b) 鉴定

    Figure  1.  Identification of vitamin A docosahexaenoate (VADHE) by HPLC (a) and MS (b)

    图  2  VADHE的纯化洗脱曲线 (a) 、全波长扫描 (b) 和核磁共振碳谱 (c) 结果

    Figure  2.  Elution curve (a), full wavelength scanning (b) and 13C NMR (c) of VADHE

    图  3  VADHE在有机溶剂体系中合成反应条件的优化

    Figure  3.  Optimization of synthetic reaction conditions in organic solvent systems of VADHE

    图  4  VADHE在无溶剂体系中合成反应条件的优化

    Figure  4.  Optimization of synthetic reaction conditions in solvent-free systems of VADHE

  • [1] ELENA C, TERENZIO C, CATERINA L, et al. Predictive role of Vitamin A serum concentration in psoriatic patients treated with IL-17 inhibitors to prevent skin and systemic fungal infections[J]. J Pharmacol Sci, 2020, 144(1): 52-56. doi: 10.1016/j.jphs.2020.06.003
    [2] ELOMDA A M, SAAD M F, SAEED A M, et al. Antioxidant and developmental capacity of retinol on the in vitro culture of rabbit embryos[J]. Zygote, 2018, 26(4): 326-332. doi: 10.1017/S0967199418000308
    [3] SHARIFIAN M, HAJIMORADLOO A, GHORBANI R, et al. Effects of dietary retinol acetate on growth performance, skin mucus immune responses and haematological parameters of Caspian roach (Rutilus caspicus)[J]. Aquac Nutr, 2017, 23(5): 893-898. doi: 10.1111/anu.12456
    [4] DOWLING J E. Vitamin A: its many roles-from vision and synaptic plasticity to infant mortality[J]. J Comp Physiol, 2020, 206(3): 389-399. doi: 10.1007/s00359-020-01403-z
    [5] 杨奇慧, 周歧存, 迟淑艳, 等. 饲料中维生素A水平对凡纳滨对虾生长、饲料利用、体组成成分及非特异性免疫反应的影响[J]. 动物营养学报, 2007, 19(6): 698-705. doi: 10.3969/j.issn.1006-267X.2007.06.009
    [6] 杨奇慧, 周小秋. 维生素A缺乏对建鲤生长性能及免疫功能的影响[J]. 中国水产科学, 2005, 12(1): 62-67. doi: 10.3321/j.issn:1005-8737.2005.01.012
    [7] 赵红霞, 崔凤杰, 李云虹, 等. 维生素酯化衍生物的合成研究进展[J]. 中国食品添加剂, 2013(6): 176-183. doi: 10.3969/j.issn.1006-2513.2013.06.023
    [8] 刘江帆, 尹春华, 王书琪. 酶法维生素改性的研究进展[J]. 食品工业科技, 2009, 30(8): 336-339.
    [9] YAO C Y, LIN W J, YUE K L, et al. Biocatalytic synthesis of vitamin A palmitate using immobilized lipase produced by recombinant Pichia pastoris[J]. Eng Life Sci, 2017, 17(7): 768-774. doi: 10.1002/elsc.201600178
    [10] AURELL M, PARRA M, TORTAJADA A, et al. Polyenolates of unsaturated carboxylic acids in synthesis. A straightforward synthesis of retinoic acids[J]. Tetrahedron lett, 1990, 31(40): 5791-5794. doi: 10.1016/S0040-4039(00)97961-9
    [11] HUANG S M, LI H J, LIU Y C, et al. An efficient approach for lipase-catalyzed synthesis of retinyl laurate nutraceutical by combining ultrasound assistance and artificial neural network optimization[J]. Molecules, 2017, 22(11): 1972.
    [12] 刘园, 倪辉, 蔡慧农, 等. 反应条件对固定化脂肪酶转酯化合成维生素A棕榈酸酯的影响研究[J]. 中国食品学报, 2012, 12(1): 75-82. doi: 10.3969/j.issn.1009-7848.2012.01.012
    [13] 高静, 姜艳军, 马丽, 等. 混合溶剂中酶促合成维生素A乳酸酯[J]. 分子催化, 2006, 20(4): 346-350. doi: 10.3969/j.issn.1001-3555.2006.04.012
    [14] MAUGARD T, REJASSE B, LEGOY M D, et al. Synthesis of water-soluble retinol derivatives by enzymatic method[J]. Biotechnol Progr, 2002, 18(3): 424-428. doi: 10.1021/bp025508f
    [15] BOHNERT K, DORIZAS A, LORENC P, et al. Randomized, controlled, multicentered, double-blind investigation of injectable poly-l-lactic acid for improving skin quality[J]. Dermatol Surg, 2019, 45(5): 718-724.
    [16] SUN G Y, SIMOUYI A, FRITSCHE K L, et al. Docosahexaenoic acid (DHA): an essential nutrient and a nutraceutical for brain health and diseases[J]. Prostag Leukotr Ess, 2018, 136: 3-13. doi: 10.1016/j.plefa.2017.03.006
    [17] ARAI E, PARMAR V M, SAHU B, et al. Docosahexaenoic acid promotes differentiation of photoreceptor cells in three-dimensional neural retinas[J]. Neurosci Res, 2017, 123: 1-7. doi: 10.1016/j.neures.2017.04.006
    [18] CHEN J N, WEI Y, CHEN X Y, et al. Polyunsaturated fatty acids ameliorate aging via redox-telomere-antioncogene axis[J]. Oncotarget, 2017, 8(5): 7301. doi: 10.18632/oncotarget.14236
    [19] SUN M J, DONG J C, XIA Y R, et al. Antibacterial activities of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) against planktonic and biofilm growing Streptococcus mutans[J]. Microb Pathog, 2017, 107: 212-218. doi: 10.1016/j.micpath.2017.03.040
    [20] SHIBABAW T. Omega-3 polyunsaturated fatty acids: anti-inflammatory and anti-hypertriglyceridemia mechanisms in cardiovascular disease[J]. Mol Cell Biochem, 2021, 476: 993-1003. doi: 10.1007/s11010-020-03965-7
    [21] ABETE P, TESTA G, GALIZIA G, et al. PUFA for human health: diet or supplementation?[J]. Curr Pharm Design, 2009, 15(36): 4186-4190. doi: 10.2174/138161209789909665
    [22] IKEDA I, IMASATO Y, NAGAO H, et al. Lymphatic transport of eicosapentaenoic and docosahexaenoic acids as triglyceride, ethyl ester and free acid, and their effect on cholesterol transport in rats[J]. Life Sci, 1993, 52(16): 1371-1379. doi: 10.1016/0024-3205(93)90172-Y
    [23] ZHANG Z, LIU F, MA X, et al. Two-stage enzymatic preparation of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) enriched fish oil triacylglycerols[J]. J Agr Food Chem, 2018, 66: 218-227. doi: 10.1021/acs.jafc.7b04101
    [24] CHOJNACKA A, GADKOWSKI W, GRUDNIEWSKA A. Lipase-catalyzed transesterification of egg-yolk phophatidylcholine with concentrate of n-3 polyunsaturated fatty acids from cod liver oil[J]. Molecules, 2017, 22: 1771.
    [25] 李金章, 王玉明, 薛长湖, 等. 脂肪酶催化乙酯甘油酯酯交换制备富含EPA和DHA的甘油三酯[J]. 中国油脂, 2011, 36(1): 13-16.
    [26] 孙兆敏, 李金章, 丛海花, 等. 酶法制备n-3多不饱和脂肪酸型磷脂的工艺[J]. 中国油脂, 2010, 35(4): 38-41.
    [27] 高泽鑫. 酶法合成维生素A棕榈酸酯[D]. 北京: 北京化工大学, 2015: 9.
    [28] 王鸿艳, 周玲妹, 张晓健, 等. 南极假丝酵母脂肪酶B的固定化及其在合成维生素A棕榈酸酯中的应用[J]. 发酵科技通讯, 2016, 45(1): 33-39. doi: 10.3969/j.issn.1674-2214.2016.01.007
    [29] 周玲妹. 南极假丝酵母脂肪酶B的生产、固定化及其在维生素A棕榈酸酯制备中的应用[D]. 杭州: 浙江工业大学, 2015: 59-60.
    [30] 冀克俭, 刘元俊, 张银生, 等. 鱼油多烯脂肪酸乙酯的NMR表征[J]. 化学分析计量, 2001, 10(3): 3-5. doi: 10.3969/j.issn.1008-6145.2001.03.002
    [31] JAHANGIRI A, MOLLER A H, DANIELSEN M, et al. Hydrophilization of bixin by lipase-catalyzed transesterification with sorbitol[J]. Food Chem, 2018, 268: 203-209. doi: 10.1016/j.foodchem.2018.06.085
    [32] NURSHAKILA M, WAHHIDA L, RAJA A R, et al. Immobilization of an Antarctic Pseudomonas AMS8 lipase for low temperature ethyl hexanoate synthesis[J]. Catalysts, 2018, 8(6): 234. doi: 10.3390/catal8060234
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出版历程
  • 收稿日期:  2021-10-08
  • 修回日期:  2021-12-15
  • 录用日期:  2021-12-30
  • 网络出版日期:  2022-01-13
  • 刊出日期:  2022-04-01

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