A multi-enzyme cascade reaction for the production of α,v-dicarboxylic acids from free fatty acids

다국어 초록 (Multilingual Abstract)

α,v-Dicarboxylic acids (α,v-DCAs) have numerous applications, as raw materials for producing variouscommodities and polymers in chemical industry. For example, dodecanedioic acid (DDDA), a monomer ofnylon 612 polymer is used for numerous applications such as plasticizers and adhesive. In this study,firstly the enzymatic production of DDDA from corresponding v-hydroxy fatty acid was optimized usingaldehyde reductase (AHR) and aldehyde dehydrogenase (ALDH), and the cofactor imbalance andproficient 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) andLactobacillus reuteri (LreNOX) were examined for the AHR/ALDH/NOX cascade and LreNOX was identifiedas suitable enzyme. Moreover, LreNOX was fused with AHR and employing this fusion protein inEscherichia. coli (E. coli) holding ALDH, nearly complete conversion (>99%) into DDDA was achieved from30 mM of 12-hydroxy dodecanoic acid within 12 h of whole-cell biotransformation (2.2-foldimprovement in productivity) turning out to be beneficial. Finally, the synthesis of DDDA fromcorresponding 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 onestep.

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