School of Energy and Chemical Engineering, UNIST, Ulsan 44919, Republic of Korea; School of Chemical and Biomolecular Engineering, Pusan National University, Geumjeong-gu, Busan 46241, Republic of Korea.
School of Energy and Chemical Engineering, UNIST, Ulsan 44919, Republic of Korea.
Bioresour Technol. 2021 Jun;329:124867. doi: 10.1016/j.biortech.2021.124867. Epub 2021 Feb 20.
Pseudomonas asiatica C1, which could grow on glucose and aerobically synthesize coenzyme B, was isolated and developed as a microbial cell factory for the production of 3-hydroxypropionic acid (3-HP) from glycerol. Three heterologous enzymes, glycerol dehydratase (GDHt), GDHt reactivase (GdrAB) and aldehyde dehydrogenase (ALDH), constituting the 3-HP synthesis pathway, were introduced, and three putative dehydrogenases, responsible for 3-HP degradation, were disrupted. In addition, the transcriptional repressor glpR and the glycerol kinase glpK were removed to increase glycerol import while eliminating the catabolic use of glycerol. Furthermore, the global regulatory protein encoded by crc and several putative oxidoreductases (PDORs) were disrupted. One resulting strain, when grown on glucose, could produce 3-HP at ~ 700 mM in 48 h in a fed-batch bioreactor experiment, with the molar yield > 0.99 on glycerol without much by-products. This study demonstrates that P. asiatica C1 is a promising host for production of 3-HP from glycerol.
从甘油生产 3-羟基丙酸（3-HP）的微生物细胞工厂的构建与开发：亚洲铜绿假单胞菌 C1 可利用葡萄糖进行有氧生长并合成辅酶 B。从甘油生产 3-羟基丙酸（3-HP）的微生物细胞工厂的构建与开发：亚洲铜绿假单胞菌 C1 可利用葡萄糖进行有氧生长并合成辅酶 B。从甘油生产 3-羟基丙酸（3-HP）的微生物细胞工厂的构建与开发：亚洲铜绿假单胞菌 C1 可利用葡萄糖进行有氧生长并合成辅酶 B。我们分离得到一株能够以葡萄糖为碳源并进行有氧合成辅酶 B 的亚洲铜绿假单胞菌 C1，并将其开发为一株从甘油生产 3-羟基丙酸（3-HP）的微生物细胞工厂。我们引入了三种异源酶，即甘油脱水酶（GDHt）、GDHt 再激活酶（GdrAB）和醛脱氢酶（ALDH），构成了 3-HP 合成途径，并敲除了三种可能负责 3-HP 降解的脱氢酶。此外，我们还去除了转录阻遏物 glpR 和甘油激酶 glpK，以增加甘油的摄取量，同时消除甘油的分解代谢利用。此外，我们还敲除了编码 crc 的全局调控蛋白和几个可能的氧化还原酶（PDORs）。在摇瓶分批培养实验中，当以葡萄糖为碳源时，一株经改造的菌株在 48 小时内可生产约 700 mM 的 3-HP，甘油的摩尔收率>0.99，且没有产生大量副产物。这项研究表明，亚洲铜绿假单胞菌 C1 是一种很有前途的从甘油生产 3-HP 的宿主菌。
