Mendes Adriano A, de Castro Heizir F, Giordano Raquel L C
Institute of Chemistry, Federal University of Alfenas, 37130-000 Alfenas, MG, Brazil; Department of Chemical Engineering, Federal University of São Carlos, 13565-905 São Carlos, SP, Brazil.
Department of Chemical Engineering, Engineering School of Lorena, University of São Paulo, 12602-810, Lorena, SP, Brazil.
Int J Biol Macromol. 2014 Sep;70:78-85. doi: 10.1016/j.ijbiomac.2014.06.035. Epub 2014 Jun 27.
The aim of this work was to prepare biocatalysts to catalyze the synthesis of butyl butyrate by esterification reaction, and the synthesis of biodiesel by transesterification of palm and babassu oils with ethanol. Lipase preparations Lipolase® (TLL1) and Lipex® 100 L (TLL2) from Thermomyces lanuginosus and Lipase AK from Pseudomonas fluorescens (PFL) were immobilized on glyoxyl-agarose beads prepared by activation with glycidol (Gly) and epichlorohydrin (Epi). The influence of immobilization time, lipase source and activating agents on the catalytic activity of the biocatalysts were evaluated in both aqueous and organic media. TLL1 immobilized on glyoxyl-agarose by 24 h of incubation resulted biocatalysts with high hydrolytic activity (varying from 1347.3 to 1470.0 IU/g of support) and thermal-stability, around 300-fold more stable than crude TLL1 extract. The maximum load of immobilized TLL1 was around 20 mg of protein/g of support. The biocatalyst prepared exhibited high activity and operational stability on the butyl butyrate synthesis by esterification after five successive cycles of 24 h each (conversion around 85-90%). Immobilized TLL1 and PFL were active in the synthesis of biodiesel by transesterification reaction. Maximum transesterification yield (≥98.5% after 48 h of reaction at 45°C) was provided by using palm oil as feedstock.
这项工作的目的是制备生物催化剂，以催化酯化反应合成丁酸丁酯，以及催化棕榈油和巴巴苏油与乙醇进行酯交换反应合成生物柴油。将来自嗜热栖热菌的脂肪酶制剂Lipolase®（TLL1）和Lipex® 100 L（TLL2）以及来自荧光假单胞菌的脂肪酶AK（PFL）固定在通过缩水甘油（Gly）和环氧氯丙烷（Epi）活化制备的乙醛酸琼脂糖珠上。在水性和有机介质中评估了固定化时间、脂肪酶来源和活化剂对生物催化剂催化活性的影响。通过24小时孵育固定在乙醛酸琼脂糖上的TLL1产生了具有高水解活性（每克载体的活性在1347.3至1470.0 IU之间）和热稳定性的生物催化剂，其稳定性比粗TLL1提取物高约300倍。固定化TLL1的最大负载量约为每克载体20毫克蛋白质。制备的生物催化剂在每次24小时的五个连续循环后，对通过酯化反应合成丁酸丁酯表现出高活性和操作稳定性（转化率约为85 - 90%）。固定化的TLL1和PFL在酯交换反应合成生物柴油中具有活性。以棕榈油为原料时，可提供最大酯交换产率（在45°C下反应48小时后≥98.5%）。
