FTIR-ATR characterization of free Rhizomucor meihei lipase (RML), Lipozyme RM IM and chitosan-immobilized RML
FTIR-ATR characterization of free Rhizomucor meihei lipase (RML), Lipozyme RM IM and chitosan-immobilized RML
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DOI：
10.1016/j.molcatb.2011.06.009
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发表时间：
2011-11-01
影响因子：
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通讯作者：
Lujan Ferreira, Maria
中科院分区：
文献类型：
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作者：
Enrique Collins, Sebastian;Lassalle, Veronica;Lujan Ferreira, Maria
以壳聚糖为载体固定化米黑根毛霉脂肪酶（RML）为催化剂，研究了其催化性能。采用两种方法将酶固定在壳聚糖上：（i）物理吸附;（ii）戊二醛共价键合。使用紫外/可见光方法分析结合在载体上的酶含量（作为可沉淀蛋白质）。采用FTIR-ATR光谱表征制备的生物催化剂，以及天然酶和商业生物催化剂Lipozyme RM IM，用作参比物质。酰胺I'信号的分析允许跟踪酶结合到支持物后的二级结构的变化及其热稳定性。通过FTIR-ATR原位监测硬脂酸乙酯的水解被用作测试反应。结果表明，RML结合Chit和Glut-Chit，其二级结构，热稳定性和酶活性在一个选定的反应测试中的微小变化。(C)2011爱思唯尔有限公司版权所有。
The synthesis and characterization of biocatalysts based on lipase from Rhizomucor miehei (RML) immobilized on chitosan-based supports were investigated. The enzyme was immobilized on chitosan following two strategies: (i) physical adsorption; and (ii) covalent bonding using glutaraldehyde. The content of enzyme bound in the supports, as precipitable protein, was analyzed using UV/visible methods. FTIR-ATR spectroscopy was employed to characterize the prepared biocatalysts, as well the native enzyme and a commercial biocatalyst Lipozyme RM IM, used as reference materials. Analysis of the amide I' signal allowed to follow the changes in the secondary structure of the enzyme after binding to the support and its thermal stability. The hydrolysis of ethyl stearate monitored in situ by FTIR-ATR was used as a test reaction. Results showed that RML was bound to Chit and Glut-Chit with minor changes in its secondary structure, thermal stability and enzymatic activity in a selected reaction test. (C) 2011 Elsevier B.V. All rights reserved.