Carro Juan, Fernández-Fueyo Elena, Fernández-Alonso Carmen, Cañada Javier, Ullrich René, Hofrichter Martin, Alcalde Miguel, Ferreira Patricia, Martínez Angel T
1Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, E-28040 Madrid, Spain.
2Department of Bio- and Environmental Sciences, International Institute Zittau - Technische Universität Dresden, Markt 23, 02763 Zittau, Germany.
Biotechnol Biofuels. 2018 Apr 2;11:86. doi: 10.1186/s13068-018-1091-2. eCollection 2018.
2,5-Furandicarboxylic acid is a renewable building block for the production of polyfurandicarboxylates, which are biodegradable polyesters expected to substitute their classical counterparts derived from fossil resources. It may be produced from bio-based 5-hydroxymethylfurfural or 5-methoxymethylfurfural, both obtained by the acidic dehydration of biomass-derived fructose. 5-Methoxymethylfurfural, which is produced in the presence of methanol, generates less by-products and exhibits better storage stability than 5-hydroxymethylfurfural being, therefore, the industrial substrate of choice.
In this work, an enzymatic cascade involving three fungal oxidoreductases has been developed for the production of 2,5-furandicarboxylic acid from 5-methoxymethylfurfural. Aryl-alcohol oxidase and unspecific peroxygenase act on 5-methoxymethylfurfural and its partially oxidized derivatives yielding 2,5-furandicarboxylic acid, as well as methanol as a by-product. Methanol oxidase takes advantage of the methanol released for in situ producing HO that, along with that produced by aryl-alcohol oxidase, fuels the peroxygenase reactions. In this way, the enzymatic cascade proceeds independently, with the only input of atmospheric O, to attain a 70% conversion of initial 5-methoxymethylfurfural. The addition of some exogenous methanol to the reaction further improves the yield to attain an almost complete conversion of 5-methoxymethylfurfural into 2,5-furandicarboxylic acid.
The synergistic action of aryl-alcohol oxidase and unspecific peroxygenase in the presence of 5-methoxymethylfurfural and O is sufficient for the production of 2,5-furandicarboxylic acid. The addition of methanol oxidase to the enzymatic cascade increases the 2,5-furandicarboxylic acid yields by oxidizing a reaction by-product to fuel the peroxygenase reactions.
2,5-呋喃二甲酸是用于生产聚呋喃二甲酸酯的可再生原料，聚呋喃二甲酸酯是有望替代源自化石资源的传统聚酯的可生物降解聚酯。它可以由生物基5-羟甲基糠醛或5-甲氧基甲基糠醛制得，这两种物质均通过生物质衍生的果糖的酸性脱水获得。在甲醇存在下生产的5-甲氧基甲基糠醛产生的副产物较少，并且比5-羟甲基糠醛具有更好的储存稳定性，因此是工业上的首选底物。
在这项工作中，已开发出一种涉及三种真菌氧化还原酶的酶促级联反应，用于从5-甲氧基甲基糠醛生产2,5-呋喃二甲酸。芳基醇氧化酶和非特异性过氧酶作用于5-甲氧基甲基糠醛及其部分氧化的衍生物，生成2,5-呋喃二甲酸以及副产物甲醇。甲醇氧化酶利用释放出的甲醇原位生成过氧化氢，该过氧化氢与芳基醇氧化酶产生的过氧化氢一起为过氧酶反应提供燃料。通过这种方式，酶促级联反应独立进行，仅需大气中的氧气作为输入，即可实现初始5-甲氧基甲基糠醛70%的转化率。向反应中添加一些外源甲醇可进一步提高产率，使5-甲氧基甲基糠醛几乎完全转化为2,5-呋喃二甲酸。
在5-甲氧基甲基糠醛和氧气存在下，芳基醇氧化酶和非特异性过氧酶的协同作用足以生产2,5-呋喃二甲酸。向酶促级联反应中添加甲醇氧化酶可通过氧化一种反应副产物为过氧酶反应提供燃料，从而提高2,5-呋喃二甲酸的产率。
