Pentadecanedioic acid production from 15‐hydroxypentadecanoic acid using an engineered biocatalyst with a co‐factor regeneration system

Abstract

α,ω-Dicarboxylic acids are valuable precursors in various chemical industries and have recently been produced using biotechnological methods to overcome the limitations of chemical synthesis. Nonanedioic acid, decanedioic acid, undecanedioic acid, and dodecanedioic acid have been produced at high concentrations from ω-hydroxycarboxylic acids using engineered biocatalysts. However, no study has been attempted on the efficient production of pentadecanedioic acid. Here, the production of pentadecanedioic acid from 15-hydroxypentadecanoic acid was carried out with alcohol dehydrogenases (ADHs), aldehyde dehydrogenases (ALDHs), and NAD(P)H oxidases, including NAD(P)H flavin oxidoreductase (NFO), that used a co-factor regeneration system, derived from several species and expressed in Escherichia coli. Among the enzymes, Kangiella koreensis ADH (KkADH), Geobacillus kaustophilus ALDH (GkALDH), and Deinococcus radiodurans NFO (DrNFO) were selected because they had the highest activity. E. coli expressing pRSF-DrNFO and pACYC-KkADH/GkALDH as the best distribution of three genes in two plasmids was used as a biocatalyst to produce pentadecanedioic acid. The optimal conditions for producing pentadecanedioic acid from 15-hydroxypentadecanoic acid by the biocatalyst were pH 8.0, 35 C, 5% (v/v) methanol, 40 g L À1 cells, and 60 mM 15-hydroxypentadecanoic acid with agitation at 250 rpm. Under these optimized conditions, 57.4 mM pentadecanedioic acid was produced after 3 h, with a molar yield of 95.6% and a productivity of 19.1 mM h À1 . The molar yield and concentration of pentadecanedioic acid showed the highest values among the reported biotechnological production of pentadecanedioic acid.

One-pot Multi-enzyme Cascade Synthesis of Bifunctional Compounds from Vegetable Oils

“…The construction of cofactor regeneration systems is not only inexpensive but also has high atom economy. A study reported that by introducing DrNFO into E. coli expressing KkADH and GtAldDH followed by optimizing the relative expression of the three genes for α,ωdicarboxylic acid production, an engineered biocatalyst with a cofactor regeneration system was successfully constructed, which was 1.6-fold more active than E. coli without cofactor regeneration [70]. NAD(P) + -dependent ADH and AldDH can form a cofactor regeneration system by coupling with NAD(P)H dehydrogenase (NOX).…”