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[Objective] To address the low yield and conversion rate in the biosynthesis of glycolic acid, we developed a whole-cell catalytic system with ethylene glycol as the substrate to obtain a strain with a high yield of glycolic acid and an efficient biosynthesis process. [Methods] With ethylene glycol as the substrate, wild-type strains were screened for their glycolic acid synthetic performance. The catalytic conditions of the optimal strain were systematically optimized to enhance the whole-cell catalytic efficiency in the production of glycolic acid. Ultraviolet mutagenesis combined with a glycolic acid biosensor-based high-throughput screening was employed to isolate high-yield strains, which were further validated in a 5-L fermenter. [Results] Pichia kudriavzevii GX-01 was successfully screened out, which could grow on glucose and produce glycolic acid from ethylene glycol via whole-cell catalysis. The whole-cell catalytic conditions were optimized by single-factor experiments as a catalytic temperature of 30 ℃, supplementation with 0.50 g MgCO3 to adjust the system pH after 6 h of reaction, a loading volume of 50 mL in a 250-mL flask, an initial glucose concentration of 25 g/L, and an initial ethylene glycol concentration of 20 g/L. Under these conditions, the glycolic acid titer in shake flasks reached 19.30 g/L after 96 h of catalysis, and the strain showed favorable tolerance to glycolic acid. A positive mutant strain P. kudriavzevii GSUV-7 was obtained through UV mutagenesis and high-throughput screening, whose glycolic acid productivity was 2.90-fold higher than that of the original strain. In a 5-L fermenter, P. kudriavzevii GSUV-7 produced 117.60 g/L of glycolic acid after 106 h of catalysis, with a 100% conversion rate. [Conclusion] This study successfully obtains a mutant strain P. kudriavzevii GSUV-7 with a high yield of glycolic acid, providing strain resources and new insights for the research on biosynthesis of glycolic acid.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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