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使用固定在硅藻土545上的AKL 13脂肪酶合成阿魏酸酯

Synthesis of Feruloyl Ester Using AKL 13 Lipase Immobilized on Celite® 545.

作者信息

Sankar Karthikumar, Achary Anant

机构信息

Department of Biotechnology, Centre for Research, Kamaraj College of Engineering and Technology,
S.P.G.C. Nagar, 625701 K. Vellakulam, Near Virudhunagar, Madurai District, Tamil Nadu, India.

出版信息

Food Technol Biotechnol. 2017 Dec;55(4):542-552. doi: 10.17113/ftb.55.04.17.5331.

DOI:10.17113/ftb.55.04.17.5331
PMID:29540988
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5848204/
Abstract

The lipophilic antioxidants, glyceryl ferulate and feruloyl glyceryl linoleate, were synthesized using lipase from AKL 13. The extracellular lipase was produced by cultivation of the strain in modified minimal medium and the enzyme was recovered by fractionation at 80% ammonium salt saturation. The concentrated enzyme with the specific activity of (4647±66) U/mg was immobilized on Celite® 545 and crosslinked using glutaraldehyde. The prepared enzyme catalyst was used for esterification of ferulic and linoleic acids with glycerol separately in hexane butane solvent system at 50 °C and 3.144× agitation. The maximum ester conversion of 94% of feruloyl glyceryl linoleate was achieved at 48 h, whereas only 35% of glyceryl ferulate was synthesized. The reaction products were characterized using RP-HPLC, FTIR, H NMR, C NMR and fluorescence spectrophotometry. The kinetic parameters of esterification reaction were determined according to ping-pong bi-bi model. The and were found to be 69.37 and 3.46 mmol, and 0.387 and 1.02 mmol/(min·g) for glyceryl ferulate and feruloyl glyceryl linoleate, respectively. The kinetic parameters were simulated in MATLAB and the experimental data were in good agreement. Furthermore, 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity of the blend of feruloyl ester and palm oil was higher than of the plain palm oil and was closer to α-tocopherol.

摘要

使用来自AKL 13的脂肪酶合成了亲脂性抗氧化剂阿魏酸甘油酯和阿魏酰甘油亚油酸酯。通过在改良的基本培养基中培养该菌株来生产胞外脂肪酶,并通过80%铵盐饱和度分级分离回收该酶。将比活性为(4647±66)U/mg的浓缩酶固定在硅藻土545上,并用戊二醛交联。制备的酶催化剂用于在50℃和3.144×搅拌条件下,分别在己烷-丁烷溶剂体系中使阿魏酸和亚油酸与甘油进行酯化反应。在48小时时,阿魏酰甘油亚油酸酯的最大酯转化率达到94%,而仅合成了35%的阿魏酸甘油酯。使用反相高效液相色谱法、傅里叶变换红外光谱法、氢核磁共振波谱法、碳核磁共振波谱法和荧光分光光度法对反应产物进行了表征。根据乒乓双底物模型确定了酯化反应的动力学参数。发现阿魏酸甘油酯和阿魏酰甘油亚油酸酯的米氏常数分别为69.37和3.46 mmol,以及0.387和1.02 mmol/(min·g)。在MATLAB中对动力学参数进行了模拟,实验数据吻合良好。此外,阿魏酰酯与棕榈油混合物的2,2-二苯基-1-苦基肼自由基清除活性高于纯棕榈油,且更接近α-生育酚。

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