α,ω-Dicarboxylic acids (α,ω-DCAs) have numerous applications, as raw materials for producing various commodities and polymers in chemical industry. For example, dodecanedioic acid (DDDA), a monomer of nylon 612 polymer is used for numerous applications such as plasticizers and adhesive. In this study, firstly the enzymatic production of DDDA from corresponding ω-hydroxy fatty acid was optimized using aldehyde reductase (AHR) and aldehyde dehydrogenase (ALDH), and the cofactor imbalance and proficient regeneration thereof, is a major hurdle for the effective biosynthesis. To circumvent this, NAD(P)H oxidases (NOXs) from Lactobacillus brevis (LbrNOX), Brevibacterium glutamicum (BreNOX) and Lactobacillus reuteri (LreNOX) were examined for the AHR/ALDH/NOX cascade and LreNOX was identified as suitable enzyme. Moreover, LreNOX was fused with AHR and employing this fusion protein in Escherichia. coli (E. coli) holding ALDH, nearly complete conversion (>99%) into DDDA was achieved from 30 mM of 12-hydroxy dodecanoic acid within 12 h of whole-cell biotransformation (2.2-fold improvement in productivity) turning out to be beneficial. Finally, the synthesis of DDDA from corresponding free fatty acid (dodecanoic acid), was performed by employing combination of AHR/ALDH/NOX cascade with CYP153AL.m, where 2.3 mM (0.53 g/L) of DDDA was produced in one-pot one step.

作为化学工业中生产各种商品和聚合物的原料，α，ω-二羧酸（α，ω-DCA）具有广泛的应用。例如，十二烷二酸（DDDA）是尼龙612聚合物的单体，可用于许多用途，例如增塑剂和粘合剂。在这项研究中，首先，使用醛还原酶（AHR）和醛脱氢酶（ALDH）优化了相应的ω-羟基脂肪酸从DDDA的酶促生产，其辅因子失衡和有效再生是有效生物合成的主要障碍。为了避免这种情况，来自短乳杆菌（LbrNOX），谷氨酸短杆菌（BreNOX）和罗伊氏乳杆菌的NAD（P）H氧化酶（NOX）检查（LreNOX）的AHR / ALDH / NOX级联，LreNOX被确定为合适的酶。此外，LreNOX与AHR融合，并在大肠杆菌中使用了该融合蛋白。持有ALDH的大肠杆菌（E. coli）， 在 全细胞生物转化后12小时内从30 mM的12-羟基十二烷酸几乎完全转化为DDDA（> 99％），结果证明是有利。最后，通过将AHR / ALDH / NOX级联与CYP153AL.m结合使用，由相应的游离脂肪酸（十二烷酸）合成DDDA，其中一锅法生产2.3  mM（0.53  g / L）DDDA。一步。