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⁻¹ 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⁻¹. The molar yield and concentration of pentadecanedioic acid showed the highest values among the reported biotechnological production of pentadecanedioic acid.

中文翻译：

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使用具有辅因子再生系统的工程生物催化剂从 15-羟基十五烷酸生产十五烷二酸

α,ω-二羧酸是各种化学工业中有价值的前体，最近已使用生物技术方法生产以克服化学合成的局限性。已经使用工程生物催化剂从ω-羟基羧酸以高浓度生产壬二酸、癸二酸、十一烷二酸和十二烷二酸。然而，尚未尝试对十五烷二酸的有效生产进行研究。在这里，使用醇脱氢酶 (ADH)、醛脱氢酶 (ALDH) 和 NAD(P)H 氧化酶，包括 NAD(P)H 黄素氧化还原酶 (NFO)，从 15-羟基十五烷酸生产十五烷二酸。一种辅因子再生系统，来源于几个物种并在大肠杆菌中表达. 在这些酶中，Kangiella koreensis ADH (KkADH)、Geobacillus kaustophilus ALDH (GkALDH) 和Deinococcus radiodurans NFO (DrNFO) 被选中，因为它们具有最高的活性。将表达 pRSF-DrNFO 和 pACYC-KkADH/GkALDH 作为三个基因在两个质粒中的最佳分布的大肠杆菌用作生产十五烷二酸的生物催化剂。生物催化剂由15-羟基十五烷酸生产十五烷二酸的最佳条件为pH 8.0, 35°C, 5% (v/v) 甲醇, 40 g L ^-1细胞和 60 mM 15-羟基十五烷酸，以 250 rpm 搅拌。在这些优化条件下，3 小时后产生了 57.4 mM 十五烷二酸，摩尔产率为 95.6%，产率为 19.1 mM h ^-1。在已报道的十五烷二酸生物技术生产中，十五烷二酸的摩尔产率和浓度显示出最高值。