纳米生物反应器内固定双酶及催化制备2,5-呋喃二甲酸的研究

2,5-呋喃二甲酸(FDCA)被美国能源部确认为未来“绿色”化学工业的一种重要平台化合物。高效制备2,5-呋喃二甲酸是研究者们的追求目标。本项目拟以5-羟甲基糠醛(HMF)为底物在纳米生物反应器内通过双酶催化氧化HMF制备FDCA。首先制备多纳米孔硅胶球载体。把重组5-羟甲基糠醛氧化酶(HMFO-ELP)和血红素硫醇过氧化物酶（HTP）次序地固定在载体的纳米孔内，形成纳米生物反应器。HMFO-ELP催化氧化HMF生成FDCA的路径为HMF→2,5-二甲酰基呋喃(DFF)→5-甲酰基-2-呋喃甲酸(FFCA)→FDCA，同时产生副产物H2O2；HTP催化氧化中间产物FFCA→FDCA，同时利用H2O2作为共底物。本项目拟研究双酶协同控制催化机制以及纳米生物反应器限定空间内底物和产物扩散传递规律及底物与酶作用机理。通过双酶协同催化，提高底物和中间产物转化率、副产物利用以及产物FDCA收率。

2,5-Furandicarboxylic acid (FDCA) is an important platform chemical. It is determined by the Department of Energy of USA as a green chemical. Efficient preparation of 2,5-Furandicarboxylic acid has been continuously perused. This project intends to synthesize 2,5-Furandicarboxylic acid form 5-hydroxymethylfurfural (HMF) under the catalysis of a two-enzyme system in nano-bioreactors. Recombinant 5-hydroxymethylfurfural oxidase (HMFO-ELP) and heme-thiolate peroxidase (HTP) are sequentially immobilized in nanoporous supports. HMFO-ELP catalyzes the oxidation of HMF to produce FDCA via following route, HMF→2,5-diformylfuran(DFF)→2,5-formylfurancarboxylic acid (FFCA)→FDCA, in the meantime H2O2 is generated. HTP catalyzes the oxidation of FFCA producing FDCA and H2O2 is used as co-substrate. This project will investigate the mechanism of coordinated catalysis by the two enzymes, the mass transfer of substrate and product within the nano-bioreactors, and the interactions between the substrate and the enzyme. It is intended to improve the efficiency of HMF conversion and the yield of FDCA under the coordinated catalysis by the two enzymes.