Fe_3O_4@SiO_2-CHO共价固定脂肪酶条件的响应面优化及固定化酶催化生产生物柴油  

Preparation of a Fe_3O_4@SiO_2-CHO carrier and immobilized lipase as a recyclable catalyst for biodiesel production

作　　者：马晓瑞[1] 秦家亮 吴海珍[1] 许毅[1] 曹瑜[1,2] 乔代蓉[1] 曹毅[1] 

机构地区：[1]四川大学生命科学学院微生物与代谢工程四川省重点实验室,成都610065 [2]四川大学生物材料工程研究中心,成都610065

出　　处：《应用与环境生物学报》2017年第2期256-263,共8页Chinese Journal of Applied and Environmental Biology

基　　金：四川省科技厅项目(2014GZX0005;2017TJ PT0001;ARRLKF14-04);国家微生物资源平台(NIMR-2016-8-1)资助~~

摘　　要：生物柴油是一种可再生且环保的化石燃料的替代品,研究固定化酶生产生物柴油对解决石化燃料耗尽和环境保护具有重要意义.将制备的纳米材料Fe_3O_4@SiO_2-CHO与伯克霍尔德氏菌脂肪酶以共价交联的方式固定,通过响应面法(RSM)优化其固定化的条件,然后以固定化酶催化橄榄油生产生物柴油,并对影响生物柴油制备的因素进行优化,最后通过GC-MS检测转酯产物的成分.结果表明:在加酶量30.22 m L、戊二醛浓度2%、固定化温度40℃、固定化时间4 h时得到最大的固定化脂肪酶酶活和固定化效率,分别为10 038 U/g和96.9%.然后以此最佳条件制得的固定化酶催化制备生物柴油,得到最优条件为加酶量1.5 g、醇油比4:1、反应温度30℃、反应时间4 h.最后用GC-MS分析所得产物,得到最大转酯效率为98.84%,且固定化酶重复转酯10次后,转酯效率仍可达90%.本研究获得的Fe_3O_4@SiO_2-CHO固定化酶不仅能在短时间内催化得到很高的生物柴油产量,且易回收、可重复使用、生产成本大大降低,是工业化生产生物柴油的一种高效环保的生物酶催化剂.Biodiesel is a renewable fuel and an alternative to conventional fossil-based diesel. Enzymatic transesterification using lipase may be used to overcome problems associated with fossil fuel depletion, including environmental issues. In this study, lipase from Burkholderia stabilis was first immobilized onto Fe3O4@SiO2-CHO nanoparticles, then the immobilization condition was optimized by response surface methodology （RSM） to obtain the most efficient immobilized lipase. Moreover, the immobilized lipase as a recyclable catalyst for biodiesel production was evaluated via transesterification of olive oil in solvent-free system. Furthermore, the fatty acid methyl esters were analysed by GC-MS to detect the catalytic ability of immobilized lipase. Under the optimal immobilization conditions： catalyst amount 30.22 mL, concentration of glutaraldehyde 2.28%, reaction temperature 40.18 ℃, reaction time 4 h, the measured activity of immobilized lipase was 10 038 U/g and the immobilized efficiency was 96.9%. The optimum conditions of transesterification were as follows： catalyst amount 1.5 g, molar ratio of methanol to oil 4 ： 1, water content 10%, reaction time 4 h, temperature 40 ℃. Under these conditions, the yield of biodiesel was 98.18% over 4 h based on the results of GC-MS, and about 90% was retained after 10 reuses. The use of Fe3O4@SiO2-CHO-immobilized lipase as a reusable catalyst for biodiesel production via transesterification greatly increases the efficiency of enzymatic biodiesel production on an industrial scale.

关 键 词：磁性纳米复合载体 Fe3O4@SiO2-CHO 脂肪酶固定化 响应面法 生物柴油 

分 类 号：Q814.2[生物学—生物工程]