A pushed biosynthesis of 2,6-dihydroxybenzoic acid by the recombinant 2,3-dihydroxybenzoic acid decarboxylase immobilized on novel amino-modified lignin-containing cellulose nanocrystal aerogel

Abstract

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 CO₂ 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 CO₂ 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.

Keywords: Biosynthesis of 2,6-dihydroxybenzoic acid; CO(2) fixation; Cellulose nanocrystal aerogel; Enzyme immobilization; Kolbe-Schmitt reaction.