Immobilization of Candida rugosa lipase on poly (3-hydroxybutyrate-co-hydroxyvalerate): a new eco-friendly support
Immobilization of Candida rugosa lipase on poly (3-hydroxybutyrate-co-hydroxyvalerate): a new eco-friendly support
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DOI：
10.1007/s10295-011-1027-3
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发表时间：
2012-02-01
影响因子：
3.4
通讯作者：
Soares, Cleide M. F.
中科院分区：
文献类型：
--
作者：
Cabrera-Padilla, Rebeca Y.;Lisboa, Milena C.;Soares, Cleide M. F.
本研究的总体目标是评价天然生物聚合物聚(3-羟基丁酸酯-羟基戊酸酯)(PHBV)在水溶液中固定化的念珠菌脂肪酶(CRL)的形态[扫描电子显微镜(SEM)]、物理化学[差示扫描量热法(DSC)、热重分析(TGA)、化学组成分析、傅立叶变换红外光谱(FTIR)、核磁共振(NMR)]和生化性质。采用物理吸附的方法对CRL进行固定化，固定化效率可达30%。与游离CRL酶相比，固定化CRL酶的活性随温度的变化略有变化(从37℃到45℃)，但最适pH值相似，为7.0。固定化CRL酶在40℃和60℃下的失活速率常数分别为0.009和0.334 h(-1)，半衰期分别为77 h和2 h。得到固定化CRL的米氏常数K-m=213.18 mm，最大反应速度V-max=318.62 U/g，并对固定化CRL的操作稳定性进行了反复试验，重复使用12次后，酶活力保持在50%以上。基于我们的研究结果，我们认为PHBV固定化的CRL可以作为一种很有前途的工业应用的生物催化剂。
The overall objective of this study is to evaluate the morphological [scanning electron microscopy (SEM)], physicochemical [differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), chemical composition analysis, Fourier-transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR)], and biochemical properties of Candida rugosa lipase (CRL) immobilized on a natural biopolymer poly(3-hydroxybutyrate-co-hydroxyvalerate) (PHBV) in aqueous solution. CRL was immobilized by physical adsorption with efficiency of 30%. Compared with free CRL enzyme, there were slight changes in immobilized CRL activity as a function of temperature (from 37 degrees C to 45 degrees C), but a similar optimal pH value of 7.0. Inactivation rate constants for immobilized CRL enzyme were 0.009 and 0.334 h(-1), and half-lives were 77 and 2 h at 40 degrees C and 60 degrees C, respectively. Kinetic parameters obtained for immobilized CRL include the Michaelis-Menten constant of K-m = 213.18 mM and maximum reaction velocity of V-max = 318.62 U/g. The operational stability of immobilized CRL was tested repeatedly, and after 12 cycles of reuse, the enzyme retained 50% activity. Based on our results, we propose that PHBV-immobilized CRL could serve as a promising biocatalyst in several industrial applications.