Jiangsu Key Laboratory for Biomass-based Energy and Enzyme Technology, Huaiyin Normal University, Huaian, China.
Jiangsu Key Laboratory for Biomass-based Energy and Enzyme Technology, Huaiyin Normal University, Huaian, China; Jiangsu Collaborative Innovation Center of Regional Modern Agriculture and Environmental Protection, Huaiyin Normal University, Huaian, China.
Bioresour Technol. 2024 Feb;394:130218. doi: 10.1016/j.biortech.2023.130218. Epub 2023 Dec 16.
Production of 2,6-dihydroxybenzoic acid (2,6-DHBA) via enzymatic carboxylation of resorcinol by decarboxylases is of great promising but shows depressed equilibrium conversion. In this study, 2,3-dihydroxybenzoic acid decarboxylase from Aspergillus oryzae (2,3-DHBD_Ao) pushing the conversion towards carboxylation for efficient 2,6-DHBA biosynthesis was achieved. Meanwhile, a novel amino-modified and lignin-doped cellulose nanocrystal aerogel (A-LCNCA) with high specific surface area and prominent CO capture was prepared for 2,3-DHBD_Ao immobilization. 2,3-DHBD_Ao@A-LCNC contributed a further enhanced conversion of carboxylation with the maximal conversion of 76.2 %, which was correlated to both the activity of 2,3-DHBD_Ao and the high CO loading capacity of A-LCNCA. Moreover, 2,3-DHBD_Ao@A-LCNC exhibited superior performances in a wider range of temperature and higher concentrations of substrate, with a prolonged storage period longer than 30 days. After seven cycles reuse, 2,3-DHBD_Ao@A-LCNCA could retain 85.3 % of its original activity. These results suggest a considerable potential of 2,3-DHBD_Ao@A-LCNCA in the selective biosynthesis of 2,6-DHBA.
通过解羧酶将间苯二酚酶促羧化为 2,6-二羟基苯甲酸(2,6-DHBA)具有很大的发展前景,但表现出平衡转化率低的问题。在本研究中,通过 2,3-二羟基苯甲酸脱羧酶(2,3-DHBD_Ao)的推动,实现了间苯二酚向羧化反应的转化,从而高效合成 2,6-DHBA。同时,制备了一种具有高比表面积和突出 CO2 捕获性能的新型氨基修饰和木质素掺杂纤维素纳米晶体气凝胶(A-LCNCA),用于 2,3-DHBD_Ao 固定化。2,3-DHBD_Ao@A-LCNCA 进一步提高了羧化转化率,最大转化率达到 76.2%,这与 2,3-DHBD_Ao 的活性和 A-LCNCA 的高 CO2 负载能力有关。此外,2,3-DHBD_Ao@A-LCNCA 在更宽的温度范围和更高的底物浓度下表现出优异的性能,储存期超过 30 天。重复使用 7 次后,2,3-DHBD_Ao@A-LCNCA 仍保留 85.3%的原始活性。这些结果表明,2,3-DHBD_Ao@A-LCNCA 在 2,6-DHBA 的选择性生物合成方面具有很大的潜力。