重组毕赤酵母发酵合成抗紫外线化合物gadusol

Recombinant Pichia pastoris Fermentation for Biosynthesis of the UV Resistant Compound Gadusol

摘要:

【目的】通过合成生物学和代谢工程技术改造毕赤酵母（Pichia pastoris，P. pastoris），实现抗紫外线化合物gadusol的高效异源合成。【方法】基于对斑马鱼gadusol生物合成途径中基因的生物信息学分析和密码子优化，通过启动子工程、代谢工程技术和发酵优化等策略，在P. pastoris中构建gadusol发酵合成途径。【结果】通过优化提取和色谱分离制备了高纯度的gadusol标准品。重组菌株P. pastoris HME的gadusol产量达到105 mg/L。通过进一步筛选基因表达启动子并优化发酵培养基组分，最终工程菌株P. pastoris GME的gadusol产量达到了约175 mg/L。【结论】该研究构建了P. pastoris GME菌株，实现了gadusol的高效合成，为加快gadusol的工业化发酵生产和在食品与化妆品中的应用研究提供了支撑和参考。

Abstract:

[Objective]This study aims to employ synthetic biology and metabolic engineering techniques to engineer Pichia pastoris (P. pastoris) for the efficient heterologous synthesis of the UV resistant compound gadusol.[Method]On the basis of a bioinformatic analysis and codon optimization of the genes involved in the gadusol biosynthetic pathway in Danio rerio, the gadusol fermentation pathway was constructed in P. pastoris via strategies such as promoter engineering, metabolic engineering, and fermentation optimization.[Result]A high-purity gadusol standard was prepared through optimized extraction and chromatographic separation. The gadusol yield in the recombinant strain P. pastoris HME reached 105 mg/L. By screening of gene expression promoters and optimization of the fermentation medium components, the engineered strain, P. pastoris GME, achieved a gadusol yield of approximately 175 mg/L.[Conclusion]This study successfully constructed the strain P. pastoris GME capable of efficiently synthesizing gadusol, providing support and a reference for accelerating the industrial production of gadusol by fermentation and the research on the application of this compound in the food and cosmetic industries.