利用葡萄糖和 1,4-丁二醇培养工程化的盐单胞菌生产聚（3-羟基丁酸-co-4-羟基丁酸酯）。
Effective production of Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by engineered Halomonas bluephagenesis grown on glucose and 1,4-Butanediol.
机构信息
School of Life Sciences, Tsinghua University, Beijing 100084, China.
School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China; Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, Guangzhou, China; Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, Guangzhou, China.
出版信息
Bioresour Technol. 2022 Jul;355:127270. doi: 10.1016/j.biortech.2022.127270. Epub 2022 May 5.
Halomonas bluephagenesis has been engineered to produce flexible copolymers P34HB or poly(3-hydroxybutyrate-co-4-hydroxybutyrate) from glucose and petrol-chemical precursor, γ-butyrolactone. Herein, gene cluster aldD-dhaT was constructed in recombinant H. bluephagenesis for catalyzing 1,4-butanediol (BDO) into 4-hydroxybutyrate, which could grow to 86 g L dry cell mass (DCM) containing 77 wt% P(3HB-co-14 mol% 4HB) in 7-L bioreactor fed with glucose and bio-based BDO. Furthermore, 4HB monomer ratio could be increased to 16 mol% by engineered H. bluephagenesis TDH4-WZY254 with defected outer-membrane. Upon deletion of 4HB degradation pathway, followed by aldD-dhaT integration, the resulted H. bluephagenesis TDB141ΔAC was grown to 95 g L DCM containing 79 wt% P(3HB-co-14 mol% 4HB) with a BDO conversion efficiency of 86% under fed-batch fermentation. Notably, 4HB molar ratio can be significantly improved to 21 mol% with negligible effects on cell growth and P34HB synthesis by adding 50% more BDO. This study successfully demonstrated a fully bio-based P34HB effectively produced by H. bluephagenesis.
海盐单胞菌已被工程化为能够利用葡萄糖和石油化学前体γ-丁内酯生产柔性共聚物 P34HB 或聚（3-羟基丁酸-co-4-羟基丁酸）。在此，aldD-dhaT 基因簇被构建在重组海盐单胞菌中以催化 1,4-丁二醇（BDO）转化为 4-羟基丁酸，该工程菌可以在 7-L 生物反应器中利用葡萄糖和生物基 BDO 生长至 86 g L 干细胞质量（DCM），其中包含 77 wt% P(3HB-co-14 mol% 4HB)。此外，通过工程化的海盐单胞菌 TDH4-WZY254 构建缺陷外膜，4HB 单体比例可以增加至 16 mol%。在缺失 4HB 降解途径后，aldD-dhaT 被整合，得到的海盐单胞菌 TDB141ΔAC 在补料分批发酵中可以生长至 95 g L DCM，其中包含 79 wt% P(3HB-co-14 mol% 4HB)，BDO 转化率为 86%。值得注意的是，通过添加 50%更多的 BDO，可以将 4HB 的摩尔比显著提高到 21 mol%，而对细胞生长和 P34HB 合成的影响可以忽略不计。本研究成功地证明了一种完全基于生物的 P34HB 可以有效地由海盐单胞菌生产。
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