Metabolic Reconstruction of Pichia pastoris for β-Carotene Biosynthesis
LI Yanru
ZHANG Xinying
LIANG Shuli
Abstract：Efficient β-carotene biosynthesis was achieved by reconstructing the metabolic network of Pichia pastoris.The synthesis pathway was established through heterologous expression of geranylgeranyl diphosphate synthase(CrtE),lycopene cyclase/phytoene synthase(CrtYB),and phytoene desaturase(CrtI)in the GS115 strain.Systematic gene screening and dosage optimization led to the development of the high-yield recombinant strain G04B,which produced 309.49 mg/L(21.71 mg/g dry cell weight[DCW])of β-carotene.Further,based on this result,β-carotene synthesis efficiency was significantly improved by enhancing the mevalonate pathway,optimizing nicotinamide adenine dinucleotide phosphate hydrogen supply,and overexpressing lipid synthesis-related genes.The optimized strain achieved a yield of 366.55 mg/L(24.34 mg/g DCW)under shake-flask conditions and 6.15 g/L(49.16 mg/g DCW)in a 5 L fermenter.The Together,the results demonstrated that optimizing the enzyme pathway,tuning gene dosage,and regulating lipid metabolism are effective strategies for increasing β-carotene production in P.pastoris,providing a foundation for natural terpenoid synthesis engineering.
Keywords：Pichia pastorisβ-carotenemetabolic reconstructioncofactor
Publication Date：2025-04-20
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：11( 141-151 )