Park Gyuyeon, Kim Ye Chan, Jang Minjeong, Park Hyuna, Lee Hong-Weon, Jeon Wooyoung, Kim Byung-Gee, Choi Kwon-Young, Ahn Jungoh
Department of Bioprocess Engineering, University of Science and Technology (UST) of Korea, Daejeon, South Korea.
School of Chemical and Biological Engineering, Seoul National University, Seoul, South Korea.
Front Bioeng Biotechnol. 2023 Jan 11;10:825576. doi: 10.3389/fbioe.2022.825576. eCollection 2022.
The α,ω-diamines (NH-(CH)-NH) and ω -amino fatty acids (NH-(CH)-COOH) have been widely used as building blocks in polymerindustries. Medium- to long-chain (C to C) fatty acid monomers with amino residues are almost exclusively produced chemical processes that generate hazardous waste and induce severe environmental problems, such as global warming and pollution. Here, we present the construction platformstrains of a cheese-ripening yeast, for direct biotransformation of hydrocarbons into medium- to long-chain α,ω-diamines and ωamino fatty acids using metabolic engineering of endogenous fatty acid ω- and β-oxidation pathways and introducing heterologous ω-transaminase in . We deleted six genes encoding the acyl-CoA oxidase (ACO1-6) and four fatty aldehyde dehydrogenase genes (FALDH1-4), which catalyze fatty acid β-oxidation and downstream oxidation of fatty aldehydes in , respectively. The ω-transaminase from DSM30191 was introduced into the genome of the Δ Δ strain under the control of -derived EXP1 promoters. The Δ Δ strains with ω-CvTA successfully accumulated the corresponding C12 αω-diamines into a shaking culture medium with dodecane or dodecanol. In addition, these strains accumulated C ω-amino fatty acids from dodecanoic acid. With the commercially available α,ω-diacid bioprocess, this yeast biosynthesis producing medium- and longchain α,ω-diamines and ω-amino fatty acids could complete the yeast platform technology generating all medium- and long-chain aliphatic polyamide monomers, α,ω-biofunctionalized with one or both carboxylic acid and amino residues.
α,ω-二胺（NH-(CH)-NH）和ω-氨基脂肪酸（NH-(CH)-COOH）已在聚合物工业中广泛用作构建模块。带有氨基残基的中长链（C至C）脂肪酸单体几乎完全通过化学方法生产，这些方法会产生有害废物并引发严重的环境问题，如全球变暖和污染。在此，我们展示了一种用于奶酪成熟的酵母的构建平台菌株，通过对内源脂肪酸ω-和β-氧化途径进行代谢工程改造，并在其中引入异源ω-转氨酶，将碳氢化合物直接生物转化为中长链α,ω-二胺和ω-氨基脂肪酸。我们删除了六个编码酰基辅酶A氧化酶（ACO1-6）的基因和四个脂肪酸醛脱氢酶基因（FALDH1-4），它们分别催化脂肪酸β-氧化和脂肪酸醛在其中的下游氧化。来自DSM30191的ω-转氨酶在源自EXP1启动子的控制下被引入ΔΔ菌株的基因组。带有ω-CvTA的ΔΔ菌株成功地在含有十二烷或十二烷醇的摇瓶培养基中积累了相应的C12αω-二胺。此外，这些菌株从十二烷酸中积累了Cω-氨基脂肪酸。利用市售的α,ω-二酸生物工艺，这种生产中长链α,ω-二胺和ω-氨基脂肪酸的酵母生物合成可以完成酵母平台技术，生成所有中长链脂肪族聚酰胺单体，这些单体通过一个或两个羧酸和氨基残基进行α,ω-双功能化。
