木质素衍生香兰素到香兰酸的 NAD+ 再生耦合酶生物转化：一种清洁生产方法

IF 5.6 1区农林科学 Q1 AGRICULTURAL ENGINEERING

Industrial Crops and Products Pub Date : 2024-10-29 DOI:10.1016/j.indcrop.2024.119921

Lingxia Xu , Sivasamy Sethupathy , Zhe Liang , Zhipeng Zhuang , Yewang Zhang , Jianzhong Sun , Daochen Zhu

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引用次数: 0

摘要

香兰素是一种著名的酚类化合物，被广泛应用于食品、香精、化妆品和制药行业，并被证明具有多种生物活性。考虑到化学合成香兰素对环境造成的负担，从木质素中提取的香兰素的酶促转化为化学合成香兰素提供了一个更绿色的替代途径。本研究利用木质素芽孢杆菌 L1 的香兰素脱氢酶（VDH）和化脓性链球菌的 NADH 氧化酶（NOX）进行生物转化，将香兰素转化为香草酸。在香兰素转化为香草酸的生物转化过程中，NOX 的加入可以从 VDH 产生的 NADH 中再生出 NAD+，从而无需继续添加昂贵的辅酶 NAD+。为了进一步提高 VDH 和 NOX 的稳定性和可重复使用性，研究人员获得了交联酶聚集体（Combi-CLVNAs），结果表明 NAD+ 的再生和香草酸的生产效果更好。在反应中批量添加香兰素后，在 31 小时内使用 2 毫摩尔 NAD+ 生产出 44.21 毫摩尔香兰素酸，分别是游离酶和不使用 NOX 时的 2.06 倍和 22.5 倍。在 4 个循环结束时，Combi-CLVNAs 总共合成了 149.98 mM 的香草酸。作为这一概念的证明，Combi-CLVNAs 成功地从木质素解聚产生的富含香兰素的提取物中生产出香兰素酸（38.69 mM）。总之，本研究提出了一种生态友好型方法，可通过 NAD+ 再生将香兰素转化为香草酸，且不会产生不需要的副产品。

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NAD+ regeneration-coupled enzymatic bioconversion of lignin-derived vanillin into vanillic acid: A cleaner production approach

Vanillic acid is a well-known phenolic compound widely used in the food, flavour, cosmetic, and pharmaceutical industries and has been shown to exhibit various biological activities. Considering the environmental burden caused by chemical synthesis of vanillic acid, the enzymatic transformation of vanillin derived from lignin offers a greener alternative to the chemical synthesis of vanillic acid offers a greener and alternative avenue. In the present study, vanillin dehydrogenase (VDH) from Bacillus ligniniphilus L1 and NADH oxidase (NOX) from Streptococcus pyogenes were used for biotransformation of vanillin into vanillic acid. The addition of NOX regenerates NAD⁺ from NADH generated by VDH during the biotransformation of vanillin into vanillic acid, which eliminates the need for continuous addition of the expensive coenzyme NAD⁺. To further improve the stability and reusability of VDH and NOX, cross-linked enzyme aggregates (Combi-CLVNAs) were obtained and the results showed better NAD⁺ regeneration and vanillic acid production. Fed-batch addition of vanillin to the reaction resulted in the production of 44.21 mM vanillic acid using 2 mM NAD⁺ in 31 h which was 2.06- and 22.5-fold higher than that of free enzymes and without NOX, respectively. A total of 149.98 mM vanillic acid was synthesized using Combi-CLVNAs at the end of 4 cycles. As a poof of this concept, Combi-CLVNAs were successfully used to produce vanillic acid (38.69 mM) from vanillin-rich extracts derived from lignin depolymerization. Overall, this study presents an eco-friendly method for transforming vanillin into vanillic acid with NAD+ regeneration, without creating unwanted by-products.

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来源期刊

Industrial Crops and Products 农林科学-农业工程

CiteScore

9.50

自引率

8.50%

发文量

1518

审稿时长

43 days

期刊介绍： Industrial Crops and Products is an International Journal publishing academic and industrial research on industrial (defined as non-food/non-feed) crops and products. Papers concern both crop-oriented and bio-based materials from crops-oriented research, and should be of interest to an international audience, hypothesis driven, and where comparisons are made statistics performed.