建立了一种新的有机相脂肪酶转酯化活性测定方法。以正己烷为溶剂，脂肪酶催化棕榈酸对硝基苯酯和正丁醇的转酯化反应为模型反应，通过测定反应液中310 nm下吸光值的变化计算反应转化率。以气相色谱法对新建的紫外分光光度法进行验证，分别采用这两种方法测定了七种商品化脂肪酶的转酯化活性，两种方法所得实验结果基本一致。利用紫外分光光度检测法考察了Lipozyme TL IM脂肪酶催化转酯化的时间进程及合成活性与酶量的关系，并对Lipozyme TL IM催化转酯化的性质(最适溶剂、酰基受体特异性、醇耐受性、最优反应温度和热力学稳定性)进行了表征。
A novel ultraviolet (UV) spectrophotometric method for evaluating lipase transesterification activi-ty in organic solvents is described. The model reaction was lipase-catalyzed transesterification between p-nitrophenyl palmitate (pNPP) and n-butanol in n-hexane at 30 °C. The conversion of pNPP to p-nitrophenol was monitored by measuring UV absorbance changes at 310 nm. Seven commercial lipases were tested using this method, and the results were similar to those obtained using gas chromatography. The UV spectrophotometric method was used to investigate the kinetics of the Lipozyme TL IM-catalyzed reaction of pNPP, and the correlation between synthetic activity and biocatalyst concentration was determined. The method was also used to investigate the Lipozyme TL IM-catalyzed transesterification properties, namely solvent effect, acyl acceptor speci-ficity, alcohol tolerance, optimum reaction temperature, and thermostability.
参考文献
| [1] | Yadav G D;Thorat P A .[J].JOURNAL OF MOLECULAR CATALYSIS B:ENZYMATIC,2012,83:16. |
| [2] | Zhu, S.-S.;Li, M.;Yu, X.;Xu, Y. .Role of Met93 and Thr96 in the Lid hinge region of rhizopus chinensis lipase[J].Applied biochemistry and biotechnology, Part A. enzyme engineering and biotechnology,2013(2):436-447. |
| [3] | Angkawidjaja C;Matsumura H;Koga Y;Takano K;Kanaya S .X-ray crystallographic and MD simulation studies on the mechanism of interfacial activation of a family I.3 lipase with two lids.[J].Journal of Molecular Biology,2010(1):82-95. |
| [4] | Ignatjev I;Valincius G;Svedaite I;Gaidamauskas E Kazemekaite M Razumas V Svendsen A .[J].Analytical Biochemistry,2005,344:275. |
| [5] | Jaeger KE;Eggert T .Lipases for biotechnology [Review][J].Current Opinion in Biotechnology,2002(4):390-397. |
| [6] | Houde A;Kademi A;Leblanc D .Lipases and their industrial applications: an overview.[J].Applied biochemistry and biotechnology, Part A. enzyme engineering and biotechnology,2004(1/3):155-170. |
| [7] | Céliz G;Martearena M R;Scaroni E;Daz M .[J].Biochemical Engineering Journal,2012,69:69. |
| [8] | Hama S;Tamalampudi S;Yoshida A;Tamadani N Kuratani N Noda H Fukuda H Kondo A .[J].Biochemical Engineering Journal,2011,55:66. |
| [9] | Yang T K;Fruekilde M B;Xu X B .[J].Journal of the American Oil Chemists'Society,2003,80:881. |
| [10] | Christensen M W;Andersen L;Husum T L;Kirk O .[J].European Journal of Lipid Science and Technology,2003,105:318. |
| [11] | Yadav G D;Lathi P S .[J].JOURNAL OF MOLECULAR CATALYSIS B:ENZYMATIC,2004,27:113. |
| [12] | Hasan, F;Shah, AA;Hameed, A .Methods for detection and characterization of lipases: A comprehensive review[J].Biotechnology Advances,2009(6):782-798. |
| [13] | Lomolino G;Di Pierro G;Lante A .[J].Food Technology and Biotechnology,2012,50:479. |
| [14] | Gupta N;Rathi P;Gupta R .[J].Analytical Biochemistry,2002,311:98. |
| [15] | Palacios, D;Busto, M. D;Ortega, N. .Study of a new spectrophotometric end-point assay for lipase activity determination in aqueous media.[J].LWT-Food Science & Technology,2014(2):536-542. |
| [16] | Torres M.;Dolcet MM.;Sala N.;Canela R. .Endophytic fungi associated with Mediterranean plants as a source of mycelium-bound lipases[J].Journal of Agricultural and Food Chemistry,2003(11):3328-3333. |
| [17] | Schmidt M;Bornscheuer U T .[J].Biomol Eng J,2005,22:51. |
| [18] | Wang B;Tang X L;Ren G F;Liu J Yu H W .[J].Biochemical Engineering Journal,2009,46:345. |
| [19] | Konarzycka-Bessler M;Bornscheuer U T .[J].ANGEWANDTE CHEMIE-INTERNATIONAL EDITION,2003,42:1418. |
| [20] | Teng Y;Xu Y .[J].Analytical Biochemistry,2007,363:297. |
| [21] | Goujard L;Villeneuve P;Barea B;Lecomte J;Pina M;Claude S;Le Petit J;Ferre E .A spectrophotometric transesterification-based assay for lipases in organic solvent[J].Analytical Biochemistry: An International Journal of Analytical and Preparative Methods,2009(1):161-167. |
| [22] | Hriscu M;Chis L;Tosa M;Irimie F D .[J].European Journal of Lipid Science and Technology,2013,115:571. |
| [23] | Soumanou MM;Bornscheuer UT .Improvement in lipase-catalyzed synthesis of fatty acid methyl esters from sunflower oil[J].Enzyme and Microbial Technology,2003(1):97-103. |
| [24] | Kim IH;Kim H;Lee KT;Chung SH;Ko SN .Lipase-catalyzed acidolysis of perilla oil with caprylic acid to produce structured lipids[J].Journal of the American Oil Chemists' Society,2002(4):363-367. |
| [25] | Romero M D;Calvo L;Alba C;Daneshfar A Ghaziaskar H S .[J].Enzyme and Microbial Technology,2005,37:42. |
| [26] | Soumanou M M;Bornscheuer U T .[J].European Journal of Lipid Science and Technology,2003,105:656. |
| [27] | Varma M N;Madras G .[J].Applied Biochemistry and Biotechnology,2010,160:2342. |
| [28] | Shintre M S;Ghadge R S;Sawant S B .[J].Journal of Chemical Technology and Biotechnology,2002,77:1114. |
| [29] | Yadav, G.D.;Pawar, S.V. .Synergism between microwave irradiation and enzyme catalysis in transesterification of ethyl-3-phenylpropanoate with n-butanol[J].Bioresource Technology: Biomass, Bioenergy, Biowastes, Conversion Technologies, Biotransformations, Production Technologies,2012(1):1-6. |
| [30] | Yadav G D;Lathi P S .[J].JOURNAL OF MOLECULAR CATALYSIS B:ENZYMATIC,2005,32:107. |
| [31] | Yadav G D;Sivakumar P .[J].Biochemical Engineering Journal,2004,19:101. |
| [32] | Talukder M M R;Das P;Fang T S;Wu J C .[J].Biochemical Engineering Journal,2011,55:119. |
| [33] | Zhang H;Pedersen LS;Kristensen D;Adler-Nissen J;Holm HC .Modification of margarine fats by enzymatic interesterification: Evaluation of a solid-fat-content-based exponential model with two groups of oil blends[J].Journal of the American Oil Chemists' Society,2004(7):653-658. |
| [34] | Khor, G.K.;Sim, J.H.;Kamaruddin, A.H.;Uzir, M.H. .Thermodynamics and inhibition studies of lipozyme TL IM in biodiesel production via enzymatic transesterification[J].Bioresource Technology: Biomass, Bioenergy, Biowastes, Conversion Technologies, Biotransformations, Production Technologies,2010(16):6558-6561. |
| [35] | Wang H;Zong MH;Wu H;Lou WY .Novel and highly regioselective route for synthesis of 5-fluorouridine lipophilic ester derivatives by lipozyme TL IM[J].Journal of Biotechnology,2007(4):689-695. |
| [36] | Barzegar A;Moosavi-Movahedi A A;Pedersen J Z;Miroliaei M .[J].Enzyme and Microbial Technology,2009,45:73. |
| [37] | Kumar A;Gross R A .[J].Biomacromolecules,2000,1:133. |
上一张
下一张
上一张
下一张
计量
- 下载量()
- 访问量()
文章评分
- 您的评分：
-
10%
-
20%
-
30%
-
40%
-
50%