水相体系中磷脂酶A1酶解南极磷虾磷脂制备溶血磷脂的分析
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(பைடு நூலகம். College of Ocean, Zhejiang University of Technology, Hangzhou 310014, China; 2. National R&D Branch Center for Pelagic Aquatic Products Processing (Hangzhou), Hangzhou 310014, China;
3. Institute of Ocean Research, Zhejiang University of Technology, Hangzhou 310032, China)
Abstract: In this study, phospholipase A1 was used to hydrolyze Antarctic krill phospholipids in aqueous media in order to obtain lysophospholipids rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The degree of phospholipid hydrolysis was measured by acid value. The composition of phospholipids was analyzed by high performance liquid chromatography (HPLC) and the acyl migration was further confirmed by measuring the content of glycerophosphatidylcholine (GPC). The fatty acid composition of phospholipids was analyzed by gas chromatographymass spectrometry (GC-MS). The results showed that the acid value increased firstly and then decreased with the increase of enzyme loading. As the hydrolysis proceeded, the content of Sn-2-lysophosphatidylcholine (Sn-2-LPC) increased initially and then reached an equilibrium value. The content of Sn-1-lysophosphatidylcholine (Sn-1-LPC) increased first and then decreased. This was due to the acyl migration of some Sn-2-LPC to form Sn-1-LPC, which was further hydrolyzed by LPA1 into glycerol phosphatidylcholine (GPC). Additionally in a short reaction period, temperature had no significant effect on the acyl migration. Finally, GC-MS analysis showed that the contents of EPA and DHA in the enzymatic hydrolysate Sn-2-LPC were relatively high, and the emulsion stability was also improved. Keywords: Antarctic krill phospholipid; lysophospholipid; phospholipase A1; acyl migration; aqueous media DOI:10.7506/spkx1002-6630-20181009-064
310014；
摘 要：采用磷脂酶A1（phospholipase A1，PLA1）在水相体系中酶解南极磷虾磷脂，以期得到富含二十碳五烯酸 （eicosapentaenoic acid，EPA）和二十二碳六烯酸（docosahexaenoic acid，DHA）的溶血磷脂。以酸价衡量磷脂酶 解程度，通过高效液相色谱对酶解前后磷脂组分进行分析，结合甘油磷脂酰胆碱（glycerol phosphatidylcholine， GPC）含量分析进一步证实酰基位移现象，并利用气相色谱-质谱分析磷脂酶解前后脂肪酸组成。结果表 明：酸价随加酶量的增加呈现先上升后下降的趋势。随着酶解时间的延长，Sn-2-溶血磷脂酰胆碱（Sn-2lysophosphatidylcholine，Sn-2-LPC）含量先增加后趋于平衡，Sn-1-溶血磷脂酰胆碱（Sn-1-lysophosphatidylcholine， Sn-1-LPC）含量先升高后降低，这是由于部分Sn-2-LPC发生酰基位移生成Sn-1-LPC，而Sn-1-LPC又进一步被PLA1 酶解生成GPC，且在较短酶解时间内，温度对酰基位移的影响不大。最后通过气相色谱-质谱分析得出酶解产物Sn2-LPC中EPA和DHA含量较高，且其乳化稳定性也得到了提高。 关键词：南极磷虾磷脂；溶血磷脂；磷脂酶A1；酰基位移；水相体系
Preparation of Lysophospholipids by Phospholipase A1-Catalyzed Hydrolysis of Antarctic Krill Phospholipids in Aqueous Phase
HU Jie1,2, YU Bokai1,2, LÜ Fei1,2, DING Yuting1,2,3, LIU Shulai1,2,3,*
92 2019, Vol.40, No.12
食品科学
※食品化学
水相体系中磷脂酶A1酶解南极磷虾磷脂制备 溶血磷脂的分析
胡 劼1,2，俞博凯1,2，吕 飞1,2，丁玉庭1,2,3，刘书来1,2,3,*
（1.浙江工业大学海洋学院，浙江 杭州 310014；2.国家远洋水产品加工技术研发分中心（杭州），浙江 杭州 3.浙江工业大学海洋研究院，浙江 杭州 310032）
3. Institute of Ocean Research, Zhejiang University of Technology, Hangzhou 310032, China)
Abstract: In this study, phospholipase A1 was used to hydrolyze Antarctic krill phospholipids in aqueous media in order to obtain lysophospholipids rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The degree of phospholipid hydrolysis was measured by acid value. The composition of phospholipids was analyzed by high performance liquid chromatography (HPLC) and the acyl migration was further confirmed by measuring the content of glycerophosphatidylcholine (GPC). The fatty acid composition of phospholipids was analyzed by gas chromatographymass spectrometry (GC-MS). The results showed that the acid value increased firstly and then decreased with the increase of enzyme loading. As the hydrolysis proceeded, the content of Sn-2-lysophosphatidylcholine (Sn-2-LPC) increased initially and then reached an equilibrium value. The content of Sn-1-lysophosphatidylcholine (Sn-1-LPC) increased first and then decreased. This was due to the acyl migration of some Sn-2-LPC to form Sn-1-LPC, which was further hydrolyzed by LPA1 into glycerol phosphatidylcholine (GPC). Additionally in a short reaction period, temperature had no significant effect on the acyl migration. Finally, GC-MS analysis showed that the contents of EPA and DHA in the enzymatic hydrolysate Sn-2-LPC were relatively high, and the emulsion stability was also improved. Keywords: Antarctic krill phospholipid; lysophospholipid; phospholipase A1; acyl migration; aqueous media DOI:10.7506/spkx1002-6630-20181009-064
310014；
摘 要：采用磷脂酶A1（phospholipase A1，PLA1）在水相体系中酶解南极磷虾磷脂，以期得到富含二十碳五烯酸 （eicosapentaenoic acid，EPA）和二十二碳六烯酸（docosahexaenoic acid，DHA）的溶血磷脂。以酸价衡量磷脂酶 解程度，通过高效液相色谱对酶解前后磷脂组分进行分析，结合甘油磷脂酰胆碱（glycerol phosphatidylcholine， GPC）含量分析进一步证实酰基位移现象，并利用气相色谱-质谱分析磷脂酶解前后脂肪酸组成。结果表 明：酸价随加酶量的增加呈现先上升后下降的趋势。随着酶解时间的延长，Sn-2-溶血磷脂酰胆碱（Sn-2lysophosphatidylcholine，Sn-2-LPC）含量先增加后趋于平衡，Sn-1-溶血磷脂酰胆碱（Sn-1-lysophosphatidylcholine， Sn-1-LPC）含量先升高后降低，这是由于部分Sn-2-LPC发生酰基位移生成Sn-1-LPC，而Sn-1-LPC又进一步被PLA1 酶解生成GPC，且在较短酶解时间内，温度对酰基位移的影响不大。最后通过气相色谱-质谱分析得出酶解产物Sn2-LPC中EPA和DHA含量较高，且其乳化稳定性也得到了提高。 关键词：南极磷虾磷脂；溶血磷脂；磷脂酶A1；酰基位移；水相体系
Preparation of Lysophospholipids by Phospholipase A1-Catalyzed Hydrolysis of Antarctic Krill Phospholipids in Aqueous Phase
HU Jie1,2, YU Bokai1,2, LÜ Fei1,2, DING Yuting1,2,3, LIU Shulai1,2,3,*
92 2019, Vol.40, No.12
食品科学
※食品化学
水相体系中磷脂酶A1酶解南极磷虾磷脂制备 溶血磷脂的分析
胡 劼1,2，俞博凯1,2，吕 飞1,2，丁玉庭1,2,3，刘书来1,2,3,*
（1.浙江工业大学海洋学院，浙江 杭州 310014；2.国家远洋水产品加工技术研发分中心（杭州），浙江 杭州 3.浙江工业大学海洋研究院，浙江 杭州 310032）