The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
Enzyme Microb Technol. 2022 Apr;155:109981. doi: 10.1016/j.enzmictec.2021.109981. Epub 2022 Jan 5.
d-Pantoic acid (D-PA) is an essential intermediate for the production of d-pantolactone. Here, three d-lactonohydrolases (D-Lacs), namely, Fm-Lac from Fusarium moniliforme SW-902, Fp-Lac from Fusarium proliferatum Nirenberg ECU2002, and Fo-Lac from Fusarium oxysporum AKU3702 were heterogeneously expressed in Pichia pastoris. The constructed recombinant strains produced D-Lacs of 1263 U/mL, 1025 U/mL, and 948 U/mL in a 3-L fermenter, respectively. Simultaneously, these three D-Lacs were used to resolve racemic pantolactone (DL-PL), the hydrolysis rate by Fo-Lac over 40% and the enantiomeric excesses was 99% after 4 h reaction, which outperformed Fm-Lac and Fp-Lac. Under the 800 mL scale reaction, the hydrolysis rate of DL-PL reached 39.2% with a D-PA concentration of 144.6 g/L and space-time yield of 36.2 g/L/h correspondingly. This is the highest catalytic efficiency reported so far, which shows that D-Lac heterologously expressed by P. pastoris has excellent industrial application prospects.
d-泛酸（D-PA）是 d-泛解酸内酯生产的必需中间产物。本文中，三种 d-内脂水解酶（D-Lacs），即来自串珠镰刀菌（Fusarium moniliforme）SW-902 的 Fm-Lac、来自扩展镰孢菌（Fusarium proliferatum）Nirenberg ECU2002 的 Fp-Lac 和来自尖孢镰刀菌（Fusarium oxysporum）AKU3702 的 Fo-Lac，在巴斯德毕赤酵母中异源表达。构建的重组菌株在 3-L 发酵罐中分别产生了 1263、1025 和 948 U/mL 的 D-Lac。同时，这三种 D-Lacs 被用于拆分外消旋泛解酸内酯（DL-PL），Fo-Lac 的水解率超过 40%，4 小时反应后对映体过量值为 99%，优于 Fm-Lac 和 Fp-Lac。在 800 mL 规模的反应中，DL-PL 的水解率达到 39.2%，D-PA 浓度为 144.6 g/L，时空产率为 36.2 g/L/h。这是迄今为止报道的最高催化效率，表明巴斯德毕赤酵母异源表达的 D-Lac 具有优异的工业应用前景。
