Furuya Toshiki, Kuroiwa Mari, Kino Kuniki
Department of Applied Chemistry, Faculty of Science and Engineering, Waseda University, 3-4-1 Ohkubo, Shinjuku-ku, Tokyo 169-8555, Japan; Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.
Department of Applied Chemistry, Faculty of Science and Engineering, Waseda University, 3-4-1 Ohkubo, Shinjuku-ku, Tokyo 169-8555, Japan.
J Biotechnol. 2017 Feb 10;243:25-28. doi: 10.1016/j.jbiotec.2016.12.021. Epub 2016 Dec 29.
Vanillin is an important and popular plant flavor, but the amount of this compound available from plant sources is very limited. Biotechnological methods have high potential for vanillin production as an alternative to extraction from plant sources. Here, we report a new approach using immobilized enzymes for the production of vanillin. The recently discovered oxygenase Cso2 has coenzyme-independent catalytic activity for the conversion of isoeugenol and 4-vinylguaiacol to vanillin. Immobilization of Cso2 on Sepabeads EC-EA anion-exchange carrier conferred enhanced operational stability enabling repetitive use. This immobilized Cso2 catalyst allowed 6.8mg yield of vanillin from isoeugenol through ten reaction cycles at a 1mL scale. The coenzyme-independent decarboxylase Fdc, which has catalytic activity for the conversion of ferulic acid to 4-vinylguaiacol, was also immobilized on Sepabeads EC-EA. We demonstrated that the immobilized Fdc and Cso2 enabled the cascade synthesis of vanillin from ferulic acid via 4-vinylguaiacol with repetitive use of the catalysts. This study is the first example of biotechnological production of vanillin using immobilized enzymes, a process that provides new possibilities for vanillin production.
香草醛是一种重要且广受欢迎的植物风味物质，但从植物来源获取的这种化合物的量非常有限。生物技术方法作为从植物来源提取香草醛的替代方法，在香草醛生产方面具有很高的潜力。在此，我们报道一种使用固定化酶生产香草醛的新方法。最近发现的加氧酶Cso2对异丁香酚和4-乙烯基愈创木酚转化为香草醛具有不依赖辅酶的催化活性。将Cso2固定在Sepabeads EC-EA阴离子交换载体上可提高操作稳定性，使其能够重复使用。这种固定化的Cso2催化剂在1mL规模下经过十个反应循环，可从异丁香酚中产生6.8mg香草醛。对阿魏酸转化为4-乙烯基愈创木酚具有催化活性的不依赖辅酶的脱羧酶Fdc也固定在Sepabeads EC-EA上。我们证明，固定化的Fdc和Cso2能够通过4-乙烯基愈创木酚从阿魏酸级联合成香草醛，且催化剂可重复使用。本研究是使用固定化酶生物技术生产香草醛的首个实例，该过程为香草醛生产提供了新的可能性。
