Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens , 9 Heroon Polytechniou Street, Zographou Campus, 15780 Athens, Greece.
J Agric Food Chem. 2017 May 3;65(17):3505-3511. doi: 10.1021/acs.jafc.7b00659. Epub 2017 Apr 19.
In the present study, the immobilization of a cutinase from Fusarium oxysporum was carried out as cross-linked enzyme aggregates. Under optimal immobilization conditions, acetonitrile was selected as precipitant, utilizing 9.4 mg protein/mL and 10 mM glutaraldehyde as cross-linker. The immobilized cutinase (imFocut5a) was tested in isooctane for the synthesis of short-chain butyrate esters, displaying enhanced thermostability compared to the free enzyme. Pineapple flavor (butyl butyrate) synthesis was optimized, leading to a conversion yield of >99% after 6 h, with an initial reaction rate of 18.2 mmol/L/h. Optimal reaction conditions were found to be 50 °C, a vinyl butyrate/butanol molar ratio of 3:1, vinyl butyrate concentration of 100 mM, and enzyme loading of 11 U. Reusability studies of imFocut5a showed that after four consecutive runs, the reaction yield reaches 54% of the maximum. The efficient bioconversion offers a sustainable and environmentally friendly process for the production of "natural" aroma compounds essential for the food industry.
在本研究中，利用戊二醛作为交联剂，通过交联酶聚集体的方法对尖孢镰刀菌来源的脂肪酶进行了固定化。在最优固定化条件下，选择乙腈作为沉淀剂，酶蛋白用量为 9.4 mg/mL，戊二醛浓度为 10 mM。固定化脂肪酶（imFocut5a）在异辛烷中用于合成短链丁酸酯，与游离酶相比，其热稳定性得到了增强。对菠萝香精（丁酸丁酯）的合成进行了优化，6 h 后转化率达到>99%，初始反应速率为 18.2 mmol/L/h。发现最佳反应条件为 50°C、丁烯酸乙烯酯/正丁醇摩尔比 3:1、丁烯酸乙烯酯浓度为 100 mM 和酶用量为 11 U。imFocut5a 的重复使用研究表明，在连续使用四次后，反应收率达到最大收率的 54%。这种高效的生物转化为食品工业中必需的“天然”香气化合物的生产提供了一种可持续且环保的方法。
