Catalytic synthesis of DHA-PC with high DHA contents by phospholipase A2 in supercritical CO2 system
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摘要: 以南极磷虾粉为原料,乙醇浸提后分离纯化得到的精制磷脂酰胆碱的纯度高达90.8%,以此为原料,在超临界CO2体系中使用磷脂酶A2（PLA2）催化磷脂酰胆碱（PC）Sn-2位脂肪酸水解,使游离DHA结合到Sn-2位以制备高DHA含量的磷脂酰胆碱（DHA-PC）,并研究了不同参数对该反应的影响。最佳酶解反应条件分别如下:固定化PLA2的添加量15%（占总底物）,时间6 h,温度55℃,压力12 MPa。在此条件下,南极磷虾磷脂酰胆碱上DHA的含量可达62.3 mol%。由此得出结论,PLA2用于磷脂酰胆碱改性以制备DHA-PC的可行性高。Abstract: With the raw material of euphausia superba powder, the purity of the refining phosphatidylcholine gained by ethyl alcohol extraction and separation and purification reached 90.8%, then the refining phosphatidylcholine was used as raw material and phospholipase A2 was used to catalyze the fatty acid hydrolysis of Sn-2 position of phosphatidylcholine in supercritical CO2 system to make the fish oil DHA to Sn-2 position and form the DHA-PC with high DHA contents.Meanwhile, the effects of different parameters on the reaction were surveyed.The results showed that, the best additive amount of immobilization phospholipase A2 was 15% ( occupied the total substratio) , the best enzymolysis was 6 h, the best enzymolysis temperature was 55 ℃, the best enzymolysis pressure was 12 MPa, respectively.Under such condition, the DHA binding ratio of euphausia superba phosphatidylcholine could reach 62.3 mol%. And it could be concluded that immobilization phospholipase A2 is highly viable for phosphatidylcholine modification to prepare DHA-PC.
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[1] Guo Z, Vikbjerg A F, Xu X.Enzymatic modification of phospholipids for functional applications and human nutrition[J].Biotechnology Advances, 2005, 23:203-259.
[2] 张思瑾.太平洋磷虾磷脂提取及磷脂酰胆碱的分离纯化研究[D].青岛:中国海洋大学, 2015. [3] Hosokawa M, Shimatani T, Kanada T, et al.Conversion to docosahexaenoic acid-containing phosphatidylserine from squid skin lecithin by phospholipase D-mediated transphosphatidylation[J].Journal of Agricultural and Food Chemistry, 2000, 48 (10) :4550.
[4] 孙甜甜.高品质南极磷虾虾油工业化生产技术研究[D].青岛:中国海洋大学, 2013. [5] Ulven S M, Kirkhus B, Lamglait A, et al.Metabolic effects of krill oil are essentially similar to those of fish oil but at lower dose of EPA and DHA, in healthy volunteers[J].Lipids, 2011, 46 (1) :37.
[6] Lin T J, Chen S W, Chang A C.Enrichment of n-3 PUFA contents on triglycerides of fish oil by lipase-catalyzed transesterification under supercritical conditions[J].Biochemical Engineering Journal, 2006, 29 (29) :27-34.
[7] Lin T J, Chen S W.Enrichment of n-3 polyunsaturated fatty acids into acylglycerols of borage oil via lipase-catalyzed reactions under supercritical conditions[J].Chemical Engineering Journal, 2008, 141 (1) :318-326.
[8] Penci M C, Constenla D T, Carelli A A.Free-fatty acid profile obtained by enzymatic solvent-free hydrolysis of sunflower and soybean lecithins[J].Food Chemistry, 2010, 120 (1) :332-338.
[9] Kamat S, Critchley G, Beckman E J, et al.Biocatalytic synthesis of acrylates in organic solvents and supercritical fluids:III.Does carbon dioxide covalently modify enzymes[J].Biotechnology&Bioengineering, 1992, 40 (1) :158-166.
[10] Garcia, Hugo S, Kim, et al.Enrichment of lecithin with n-3fatty acids by acidolysis using immobilized phospholipase A1[J].Grasas Y Aceites, 2008, 59 (4) :368-374.
[11] 张潇予.南极磷虾磷脂酰胆碱的提纯[D].济南:山东师范大学, 2012. [12] Egger D, Wehtje E, Adlercreutz P.Characterization and optimization of phospholipase A2catalyzed synthesis of phosphatidylcholine[J].Biochimica Et Biophysica Acta, 1997, 1343 (1) :76-84.
[13] Kim I H, Garcia H S, Jr C G H.Phospholipase A-catalyzed synthesis of phospholipids enriched in polyunsaturated fatty acid residues[J].Enzyme&Microbial Technology, 2007, 40 (5) :1130-1135.
[14] Kim I H, Garcia H S, Hill C G.Synthesis of Structured Phosphatidylcholine Containing n-3 PUFA Residues via Acidolysis Mediated by Immobilized Phospholipase A1[J].Journal of the American Oil Chemists’Society, 2010, 87 (11) :1293-1299.
[15] Vikbjerg A F, Mu H, Xu X.Synthesis of structured phospholipids by immobilized phospholipase A2catalyzed acidolysis[J].Journal of Biotechnology, 2007, 128:545-554.
[16] 李响.磷脂酶A1的固定化及其催化合成DHA/EPA型磷脂的研究[D].广州:华南理工大学, 2015. [17] Kim J, Lee C S, Oh J, et al.Production of egg yolk lysolecithin with immobilized phospholipase A2[J].Enzyme&Microbial Technology, 2001, 29 (10) :587-592.
[18] 韩轶, 周扬, 赵永芳.卵磷脂的提纯、鉴定及应用[J].氨基酸和生物资源, 2001, 23:28-31. [19] 胡小中, 温光源, 龙金林.磷脂分析检测技术研究进展[J].中国油脂, 2003, 28:41-44. [20] Xi X, Feng X, Shi N, et al.Immobilized phospholipase A1-catalyzed acidolysis of phosphatidylcholine from Antarctic krill (Euphausia superba) for docosahexaenoic acid enrichment under supercritical conditions[J].Journal of Molecular Catalysis B Enzymatic, 2016, 126:46-55.
[21] Dufour E, HervéG, Haertle T.Hydrolysis ofβ-lactoglobulin by thermolysin and pepsin under high hydrostatic pressure[J].Biopolymers, 2004, 35 (5) :475-483.
[22] Gross M, Auerbach G, Jaenicke R.The catalytic activities of monomeric enzymes show complex pressure dependence[J].Febs Letters, 1993, 321 (2-3) :256-260.
[23] Almeida M C, Rui R, Maia C, et al.Novozym 435 Activity in Compressed Gases.Water Activity and Temperature Effects[J].Enzyme&Microbial Technology, 1998, 22 (6) :494-499.
[24] Peres C, Harper N, Silva M D R G D, et al.Effect of zeolites on lipase catalyzed esterification in nonaqueous media[J].Enzyme&Microbial Technology, 2005, 37 (1) :145-149.
[25] 朱珊珊.磷脂酶A1的固定化及催化合成富含共轭亚油酸磷脂的研究[D].广州:华南理工大学, 2011. [26] Fornari T, Stateva R P.High Pressure Fluid Technology for Green Food Processing[M].Springer International Publishing, 2015.
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