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Coupling a recombinant oxidase to catalase through specific noncovalent interaction to improve the oxidation of 5-hydroxymethylfurfural to 2,5-furandicarboxylic acid
Enzyme and Microbial Technology ( IF 3.4 ) Pub Date : 2021-08-13 , DOI: 10.1016/j.enzmictec.2021.109895
Yining Han 1 , Wenxin Qu 1 , Wei Feng 1
Affiliation  

5-Hydroxymethylfurfural oxidase (HMFO) can catalyze both hydroxyl and aldehyde oxidations. It catalyzes 5-hydroxymethylfurfural into 2,5-furandicarboxylic acid. However, the application of HMFO encountered two problems: the expressed HMFO in Escherichia coli. is largely in the form of inclusion bodies, and the by-product of H2O2 has a negative effect on HMFO stability. To solve these problems, recombinant HMFO was generated by fusing the C-terminus to an elastin-like polypeptide (ELP). ELP-HMFO can be expressed with significantly reduced inclusion bodies. ELP-HMFO exhibited improved stability and tolerance toward H2O2. Further recombination is carried out by fusing the N-terminus of HMFO to a glutamic acid-rich leucine zipper motif (ZE). Similarly, recombinant catalase (CAT) is generated by fusing the N-terminus to ELP and fusing the C-terminus to an arginine-rich leucine zipper motif (ZR). ELP-HMFO-ZE can interact specifically with ZR-CAT-ELP, ascribing to the coiled-coil association of ZE and ZR. ELP-HMFO-ZE#ZR-CAT-ELP coordinates the respective catalytic activities of the two enzymes. ELP-HMFO-ZE catalyzes the oxidation of HMF, and the generated hydrogen peroxide is decomposed by ZR-CAT-ELP into H2O and oxygen. During the oxidation of HMF, the cofactor FAD of HMFO is reduced, and molecular oxygen is needed to reoxidize the reduced FAD. The evolved oxygen from the decomposing of H2O2 can just meet the requirement, which can be diffused efficiently from ZR-CAT-ELP to ELP-HMFO-ZE due to the short distance between the two enzymes.



中文翻译:

通过特定的非共价相互作用将重组氧化酶与过氧化氢酶偶联,以改善 5-羟甲基糠醛氧化为 2,5-呋喃二甲酸

5-羟甲基糠醛氧化酶 (HMFO) 可以催化羟基和醛氧化。它催化 5-羟甲基糠醛生成 2,5-呋喃二甲酸。然而,HMFO的应用遇到了两个问题:在大肠杆菌中表达的HMFO 。主要以包涵体的形式存在,H 2 O 2的副产物对 HMFO 的稳定性有负面影响。为了解决这些问题,通过将 C 端与弹性蛋白样多肽 (ELP) 融合来产生重组 HMFO。ELP-HMFO 可以用显着减少的包涵体表达。ELP-HMFO 对 H 2 O 2表现出更高的稳定性和耐受性. 进一步重组是通过将 HMFO 的 N 端融合到富含谷氨酸的亮氨酸拉链基序 (Z E ) 来进行的。类似地,通过将​​ N 端融合到 ELP 并将 C 端融合到富含精氨酸的亮氨酸拉链基序 (Z R )来产生重组过氧化氢酶 (CAT )。ELP-HMFO-Z E可以与 Z R -CAT-ELP特异性相互作用,归因于 Z E和 Z R的盘绕线圈关联。ELP-HMFO-Z E # Z R -CAT-ELP 协调两种酶各自的催化活性。ELP-HMFO-Z E催化HMF氧化,生成的过氧化氢被Z R分解-CAT-ELP 转化为 H 2 O 和氧气。在HMF氧化过程中,HMFO的辅因子FAD被还原,需要分子氧来重新氧化还原的FAD。H 2 O 2分解放出的氧刚好满足要求,由于两种酶之间的距离短,可以有效地从Z R -CAT-ELP扩散到ELP-HMFO-Z E。

更新日期:2021-08-20
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