固定化脂肪酶催化酱油糟油脂合成甘油二酯的研究
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摘要
本文以酱油糟为研究对象,利用实验室自主研发的低温连续相变萃取技术提取其中的油脂,并对提取出来的油脂主要理化指标进行分析,选用自制的具有Sn-1,3位专一性固定化脂肪酶HM IM对其油脂中丰富的游离脂肪酸(含量为61.50%)酯化合成甘油二酯。通过酶的添加量、甘油添加量、反应温度、搅拌速度、金属离子和不同辅助脱酸处理这几个因素进行优化,得到最佳的酯化合成甘油二酯条件为:自制固定化脂肪酶00S添加量1%、甘油添加量为理论所需量的1.2倍、反应温度65℃、真空度0.098 MPa+氮吹,2 mmol K+处理固定化酶、搅拌速度120 r/min下反应10 h,酱油糟油脂的游离脂肪酸酯化率达96.36%,甘油二酯含量达48.65%(43.65%的1,3-DG、5.00%的1,2-DG)、2.24%FFA、9.31%的MG,39.80%的TG,而且酯化反应不会改变油脂的脂肪酸组成。酶法酯化处理既降低了酱油糟油脂的酸值、又富含功能性甘油二酯,对其开发再利用提供新的途径。
Soy sauce residue oil was was extracted with free fatty acids content of 61.50% by a novel continuous phase transition extraction method at low temperature,and the self-made immobilized lipase HMIM with Sn-1,3 specificity was used to catalyze the synthesis of diacylglycerol.The enzyme dosage the content of glycerin,reaction temperature,metal ions and auxiliary acid treatment methods were optimized,and the optimal conditions were determined as follows:immobilized lipase dosage 1%,content of glycerol 1.2 times of the required,reaction temperature:65 ℃,vacuum degree;0.098 MPa,K+ concentration 2 mmo L,stirring speed 20 r/min and reaction time 10 h.Under these conditions,the free fatty acid esterification rate was 96.36% and the content of diglyceride was 48.65%(43.65% of 1,3-DG,5.00% of 1,2-DG).The contents of FFA,MG and TG were 2.24%,9.31% and 39.80%,respectively.Results showed that esterification reaction would not change the fatty acid composition of soy sauce residue oil.Process of Enzymatic esterification not only reduced the acid value of soy sauce residue oil but also increased the contents of functional diacylglycerol,which provided a new method for the development and utilization of soy sauce residue.
Soy sauce residue oil was was extracted with free fatty acids content of 61.50% by a novel continuous phase transition extraction method at low temperature,and the self-made immobilized lipase HMIM with Sn-1,3 specificity was used to catalyze the synthesis of diacylglycerol.The enzyme dosage the content of glycerin,reaction temperature,metal ions and auxiliary acid treatment methods were optimized,and the optimal conditions were determined as follows:immobilized lipase dosage 1%,content of glycerol 1.2 times of the required,reaction temperature:65 ℃,vacuum degree;0.098 MPa,K+ concentration 2 mmo L,stirring speed 20 r/min and reaction time 10 h.Under these conditions,the free fatty acid esterification rate was 96.36% and the content of diglyceride was 48.65%(43.65% of 1,3-DG,5.00% of 1,2-DG).The contents of FFA,MG and TG were 2.24%,9.31% and 39.80%,respectively.Results showed that esterification reaction would not change the fatty acid composition of soy sauce residue oil.Process of Enzymatic esterification not only reduced the acid value of soy sauce residue oil but also increased the contents of functional diacylglycerol,which provided a new method for the development and utilization of soy sauce residue.
引文
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[19]韩抒.高酸值米糠油酯化脱酸工艺的研究[D].合肥:合肥工业大学,2009HAN Shu.Esterifical refine technics of high acid value of rice bran oil[D].Hefei:Hefei University of Technology,2009
[20]Yomi W,Yoshie Y,Toshihoro N,et al.Production of mono-acylglycerol of conjugated linoleic acid by esterification followed by dehydration at low temperature using Penicillium camembertii lipase[J].Journal of Molecular Catalysis B:Enzymatic,2004,27:249-254
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[2]卜春文.酱油渣的生物技术开发利用[J].饲料研究,2001,22(12):49-50BU Chun-wen.The biotechnology development and utilization of soy sauce residue[J].Feed Research,2001,22(12):49-50
[3]ZHAO Li-chao,ZHANG Yong,HE Li-ping,et al.Soy Sauce residue oil extracted by a novel continuous phase transition extraction under low temperature and its refining process[J].Journal of Agricultural and Food Chemistry,2014,62(14):3230-3235
[4]阎杰,宋光泉.值得开发的“废物”-酱油渣[J].中国调味品,2006,10:14-17YAN Jie,SONG Guang-quan.Soy sauce residue-a kind of industrial garbage being worth studying[J].China Condiment,2006,10:14-17
[5]杨莹莹,姚舜,万海清,等.酱油渣中油脂提取工艺的研究及其脂肪酸成分分析[J].中国酿造,2011,30(11):174-177YANG Ying-ying,YAO Shun,WAN Hai-qing,et al.Fat extraction from soy sauce residues and fatty acids composition analysis[J].China Brewing,2011,30(11):174-177
[6]钟振声,王亚丽,齐玉堂,等.酱油渣中油脂的提取及其质量指标分析[J].中国油脂,2006,31(11):21-23ZHONG Zhen-sheng,WANG Ya-li,QI Yu-tang,et al.Analysis on the extraction of oil from soy sauce residue and its quality index[J].China Grease,2006,31(11):21-23
[7]刘志伟,况伟,杨钦阶,等.酱油渣中提取油脂和黄酮[J].嘉应学院学报,2012,30(11):66-70LIU Zhi-wei,KUANG Wei,YANG Qin-jie,et al.Extraction of oil and flavonoids from soy sauce residue[J].Journal of Jiaying University,2012,30(11):66-70
[8]李脉,张松涛,杨继国,等.酱油渣中油脂的亚临界流体提取及酶法脱酸的研究[J].农业机械,2011,20:52-55LI Mai,ZHANG Song-tao,YANG Ji-guo,et al.Study on subcritical fluid extraction and enzymatic deacidification of oil in soy sauce[J].Farm Machinery,2011,20:52-55
[9]Flickinger B D,Noboru M.Nutritional characteristics of DAG oil[J].Lipids,2003,38(2):129-132
[10]Osamu M,Yasushi T,Jeannie B K,et al.Safety assessment of heated diacylglycerol oil:sub-chronic toxicity study in rats[J].Food and Chemical Toxicology,2008,46(8):2748-2757
[11]Yasukawa T.Nutritional functions of dietary diacylglycerols[J].Journal of Oleo Science,2001,50(5):427-432
[12]董攀,李鑫,李丹丹,等.响应曲面法优化Lipozyme TL IM催化大豆油制备1,3-甘油二酯[J].食品工业科技,2013,34(7):190-194,198DONG Pan,LI Xin,LI Dan-dan,et al.Optimization of production condition of diacylglycerol from soybean oil hydrolyzed by lipozyme TL IM by response surface methodology[J].Science and Technology of Food Industry,2013,34(7):190-194,198
[13]邹冬芽.甘油二酯的酶法制备及其功能性研究[D].无锡:江南大学,2004ZHOU Dong-ya.Study on enzymatic preparation and functional study of glyceride diester[D].Wuxi:Jiangnan University,2004
[14]胡士恒.玉米油酶法合成甘油二酯研究[D].长春:吉林大学,2013HU Shi-heng.Study on enzymatic synthesis of diglycerides by corn oil[D].Changchun:Jilin University,2013
[15]刘小如,张超,胡蒋宁,等.酶催化茶油合成甘油二酯工艺及脂肪酸变化[J].食品科学,2013,34(2):36-41LIU Xiao-ru,ZHANG Chao,HU Jiang-ning,et al.Lipase-catalyzed synthesis of diacylglycerol from camellia oil and fatty acid profile change[J].Food Science,2013,34(2):36-41
[16]曹庸.一种连续相变萃取酱油渣油脂的方法:中国,CN103173280A[P]2013-06-26CAO Yong.A continuous phase transition extraction method of soy sauce residue oil:China,CN103173280A[P]2013-06-26
[17]FUCHS Beate,SüSS Rosmarie,TEUBER Kristin,et al.Lipid analysis by thin-layer chromatography-a review of the current state[J].Journal of Chromatography A,2011,1218(19):2754-2774
[18]鲍方宇,张敏.薄层色谱法测定油脂中甘油三酯含量[J].食品科学,2013,34(4):125-128BAO Fang-yu,ZHANG Min.Determination of triglyceride content in oils and fats by thin layer chromatography[J].Food Science,2013,4(34):125-128
[19]韩抒.高酸值米糠油酯化脱酸工艺的研究[D].合肥:合肥工业大学,2009HAN Shu.Esterifical refine technics of high acid value of rice bran oil[D].Hefei:Hefei University of Technology,2009
[20]Yomi W,Yoshie Y,Toshihoro N,et al.Production of mono-acylglycerol of conjugated linoleic acid by esterification followed by dehydration at low temperature using Penicillium camembertii lipase[J].Journal of Molecular Catalysis B:Enzymatic,2004,27:249-254
[21]胡隼,王芳,谭天伟.固定化假丝酵母99-125脂肪酶催化合成甘油二酯[J].北京化工大学学报(自然科学版),2007,34(2):196-199,210HU Sun,WANG Fang,TAN Tian-wei.Synthesis of diglycerides by enzymatic esterification using immobilized lipase from Candida sp.99-125 in a solvent-free system[J].Journal of Beijing University of Chemical Technology(Natural Science Edition),2007,34(2):196-199,210