In this study, 2-monoacyl glycerolrich in 2D-MAG was synthesized from algal oil by ethanol, and the types of lipase, the molar ratio of algal oil to ethanol, reaction time, reaction temperature and utilization times of lipase were studied. The results showed that n-3 PUFA (62.26%) was the most abundant fatty acid of sn-2 in algal oil, and it was an ideal raw material for the synthesis of the sn-2 long-chain structural lipid 2D-MAG.The catalytic activity of lipozyme RM IM >Novozym 435 >lipozyme TL IM >Newlase F > Lipase AK > Lipase AY was in the order of high to low. Lipozyme RM IM was used to catalyze the alcoholysis of alga oil under the conditions of molar ratio of alga oil/ethanol of 1∶40, enzyme addition of 10%, reaction time of 3h and reaction temperature of 35 ℃, which was more conducive to the accumulation of 2D-MAG synthesis products. Under these conditions, the yield of 2D-MAG was 45.36%. After the 6th repetition of Lipozyme RM IM lipase, the content of 2D-MAG was significantly lower than that of the previous five times (P<0.05), indicating that although the overall catalysis of Lipozyme RM IM lipase was stable, its catalytic activity significantly weakened with the increase of repeated use. The results will provide references for the preparation and modification of functional structural lipids of related sn-2 long-chain polyunsaturated fatty acids.
WANG Qiang
,
XIE Yuejie
,
LI Yuanyuan
,
WEI Huaheng
,
HE Zhifei
,
LI Hongjun
. Preparation of sn-2 long-chain polyunsaturated fatty acid monoglyceride and its influencing factors[J]. Food and Fermentation Industries, 2020
, 46(10)
: 19
-26
.
DOI: 10.13995/j.cnki.11-1802/ts.023618
[1] HU P, XU X, YU L L. Interesterified trans-free fats rich in sn-2 nervonic acid prepared using Acer truncatum oil, palm stearin and palm kernel oil, and their physicochemical properties[J]. LWT - Food Science and Technology, 2017, 76:156-163.
[2] ARMAND M. Milk fat digestibility[J]. Sciences des Aliments, 2008, 28:84-98.
[3] DOMENICHIELLO A F, KITSON A P, BAZINET R P. Is docosahexaenoic acid synthesis from α-linolenic acid sufficient to supply the adult brain[J]. Progress in Lipid Research, 2015, 59:54-66.
[4] PFEFFER J, FREUND A, BEL-RHLID R, et al. Highly efficient enzymatic synthesis of 2-monoacylglycerides and structured lipids and their production on a technical scale[J]. Lipids, 2007, 42(10):947-953.
[5] MICHALSKI M C, GENOT C, GAYET C, et al. Multiscale structures of lipids in foods as parameters affecting fatty acid bioavailability and lipid metabolism[J]. Progress in Lipid Research, 2013, 52(4):354-373.
[6] TANG W J, WANG X S, HUANG J H,et al.A novel method for the synthesis of symmetrical triacylglycerols by enzymatic transesterification[J]. Bioresource Technology, 2015, 196:559-565.
[7] MUNIO M, ROBLES A, ESTEBAN L, et al. Synthesis of structured lipids by two enzymatic steps: Ethanolysis of fish oils and esterification of 2-monoacylglycerols[J]. Process Biochemistry, 2009, 44(7):723-730.
[8] TURON F, BACHAIN P, CARO Y, et al. A direct method for regiospecific analysis of TAG using α-MAG[J]. Lipids, 2002, 37(8):817-821.
[9] WANG X, LI M, WANG T, et al. An improved method for the synthesis of 2-arachidonoylglycerol[J]. Process Biochemistry, 2014, 49(9):1 415-1 421.
[10] ESTEBAN L, MARÍA DEL MAR MUÍO, ROBLES A, et al. Synthesis of 2-monoacylglycerols (2-MAG) by enzymatic alcoholysis of fish oils using different reactor types[J]. Biochemical Engineering Journal, 2009, 44(2-3):271-279.
[11] RODRÍGUEZ A, ESTEBAN L, MARTÍN L, et al. Synthesis of 2-monoacylglycerols and structured triacylglycerols rich in polyunsaturated fatty acids by enzyme catalyzed reactions[J]. Enzyme and Microbial Technology, 2012, 51(3):148-155.
[12] JOSÉ L G. Stearidonic acid (18:4n-3): Metabolism, nutritional importance, medical uses and natural sources[J]. European Journal of Lipid Science and Technology, 2007, 109: 1 226-1 236.
[13] MARÍA M M, ESTEBAN L, ROBLES A, et al. Synthesis of 2-monoacylglycerols rich in polyunsaturated fatty acids by ethanolysis of fish oil catalyzed by 1,3 specific lipases[J]. Process Biochemistry, 2008, 43(10):1 033-1 039.
[14] PIYATHEERAWONG W, IWASAKI Y, XU X, et al. Dependency of water concentration on ethanolysis of trioleoylglycerol by lipases[J]. Journal of Molecular Catalysis B Enzymatic, 2004, 28(1):19-24.
[15] ESTEBAN L, MARÍA J J, HITA E, et al. Production of structured triacylglycerols rich in palmitic acid at sn-2 position and oleic acid at sn-1,3 positions as human milk fat substitutes by enzymatic acidolysis[J]. Biochemical Engineering Journal, 2011, 54(1):62-69.
[16] HAN L, XU Z, HUANG J, et al. Enzymatically catalyzed synthesis of low-calorie structured lipid in a solvent-free system: Optimization by response surface methodology[J]. Journal of Agricultural and Food Chemistry, 2011, 59(23):12 635-12 642.
[17] IRIMESCU R, IWASAKI Y, HOU C T. Study of TAG ethanolysis to 2-MAG by immobilizedCandidaantarcticalipase and synthesis of symmetrically structured TAG[J]. Journal of the American Oil Chemists' Society, 2002, 79(9):879-883.
[18] ZHANG Y, WANG X, ZOU S, et al. Synthesis of 2-docosahexaenoylglycerol by enzymatic ethanolysis[J]. Bioresource Technology, 2018, 251:334-340.
[19] HE Y, LI J, KODALI S, et al. Liquid lipases for enzymatic concentration of n-3 polyunsaturated fatty acids in monoacylglycerols via ethanolysis: Catalytic specificity and parameterization[J]. Bioresource Technology, 2017, 224:445-456.
[20] ZHANG Y, WANG X, XIE D, et al. Synthesis and concentration of 2-monoacylglycerols rich in polyunsaturated fatty acids[J]. Food Chemistry, 2018, 250:60-66.
[21] PIYATHEERAWONG W, IWASAKI Y, XU X, et al. Dependency of water concentration on ethanolysis of trioleoylglycerol by lipases[J]. Journal of Molecular Catalysis B Enzymatic, 2004, 28(1):19-24.
[22] IKEDA I, SASAKI E, YASUNAMI H, et al. Digestion and lymphatic transport of eicosapentaenoic and docosahexaenoic acids given in the form of triacylglycerol, free acid and ethyl ester in rats[J]. Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1995, 1259(3):297-304.
[23] DUAN Z Q, DU W, LIU D H. The solvent influence on the positional selectivity of Novozym 435 during 1,3-diolein synthesis by esterification[J]. Bioresour Technol, 2010, 101(7):2 568-2 571.
