Division of Chemical Engineering and Materials Science, Ewha Womans University, 52 Ewhayeodae-gil, Seodaemun-gu, Seoul 03760, Republic of Korea.
Bio-based Chemistry Research Center, Advanced Convergent Chemistry Division, Korea Research Institute of Chemical Technology, P.O. Box 107, 141 Gajeong-ro, Yuseong-gu, Daejeon 34602, Republic of Korea.
Int J Biol Macromol. 2020 Apr 15;149:593-599. doi: 10.1016/j.ijbiomac.2020.01.254. Epub 2020 Jan 27.
Sucrose utilization has been established in Escherichia coli strains by expression of Mannheimia succiniciproducens β-fructofuranosidase (SacC), which hydrolyzes sucrose into glucose and fructose. Recombinant E. coli strains that can utilize sucrose were examined for their abilities to produce poly(3-hydroxybutyrate) [P(3HB)] and poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)] from sucrose. When recombinant E. coli strains expressing Ralstonia eutropha PhaCAB and SacC were cultured in MR medium containing 20 g/L of sucrose, all recombinant E. coli strains could produce P(3HB) from sucrose. Also, recombinant E. coli strains expressing Pseudomonas sp. MBEL 6-19 PhaC1437, Clostridium propionicum Pct540, R. eutropha PhaAB enzymes along with SacC could produce P(3HB-co-LA) from sucrose. Among the examined E. coli strains, recombinant E. coli XL1-Blue produced the highest contents of P(3HB) (53.60 ± 2.55 wt%) and P(3HB-co-LA) (29.44 ± 0.39 wt%). In the batch fermentations, recombinant E. coli XL1-Blue strains completely consumed 20 g/L of sucrose as the sole carbon source and supported the production of 3.76 g/L of P(3HB) and 1.82 g/L of P(3HB-co-LA) with 38.21 wt% P(3HB) and 20.88 wt% P(3HB-co-LA) contents, respectively. Recombinant E. coli strains developed in this study can be used to establish a cost-efficient biorefinery for the production of polyhydroxyalkanoates (PHAs) from sucrose, which is an abundant and inexpensive carbon source.
在大肠杆菌菌株中通过表达曼海姆西亚糖醛酸梭菌β-呋喃果糖苷酶（SacC）来建立蔗糖利用，SacC 将蔗糖水解成葡萄糖和果糖。研究了能够利用蔗糖的重组大肠杆菌菌株从蔗糖生产聚（3-羟基丁酸酯）[P（3HB）]和聚（3-羟基丁酸酯-共-乳酸酯）[P（3HB-co-LA）]的能力。当在含有 20 g/L 蔗糖的 MR 培养基中培养表达罗尔斯通氏菌 PhaCAB 和 SacC 的重组大肠杆菌菌株时，所有重组大肠杆菌菌株都可以从蔗糖中生产 P（3HB）。此外，表达假单胞菌 MBEL 6-19 PhaC1437、丙酸梭菌 Pct540、罗尔斯通氏菌 PhaAB 酶和 SacC 的重组大肠杆菌菌株可以从蔗糖生产 P（3HB-co-LA）。在所检查的大肠杆菌菌株中，重组大肠杆菌 XL1-Blue 产生的 P（3HB）（53.60±2.55 wt%）和 P（3HB-co-LA）（29.44±0.39 wt%）含量最高。在分批发酵中，重组大肠杆菌 XL1-Blue 菌株完全消耗 20 g/L 的蔗糖作为唯一碳源，并支持生产 3.76 g/L 的 P（3HB）和 1.82 g/L 的 P（3HB-co-LA），分别具有 38.21 wt% P（3HB）和 20.88 wt% P（3HB-co-LA）的含量。本研究开发的重组大肠杆菌菌株可用于建立从蔗糖生产聚羟基烷酸酯（PHA）的经济型生物精炼厂，蔗糖是一种丰富且廉价的碳源。
