Abstract
Purpose
Pichia pastoris is well known for its ability to produce short and low-immunogenic humanized glycosyl chains onto recombinant glycoproteins, it was thus speculated to be applicable to synthesize oligosaccharides. In this study, generally recognized as safe (GRAS) microorganism Pichia pastoris GS115 was tested for its potential to be used as a new synthetic chassis to produce the most abundant human milk oligosaccharide 2′-fucosyllactose (2′-FL).
Methods
To enable the de novo synthesis of 2′-FL, lactose transporter lac12, two enzymes of gmd, gmer, and fucosyltransferases futC were integrated into the genome of P. pastoris, under the control of constitutive PGAP promoter.
Results
The resulting recombinant yeasts yielded up to 0.276 g/L through culture optimization in a 5 L bioreactor.
Conclusion
To our knowledge, this is the first report of 2′-FL production in engineered Pichia pastoris. This work is a good starting point to produce 2′-FL using Pichia pastoris as a viable chassis.
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Acknowledgements
This research was supported by the National Key R&D Program of China (Grant Number 2019YFD0901800), Shanghai Post-doctoral Excellence Program (2020132), Natural Science Foundation of Shanghai (22ZR1418200) and the 111 Project (B18022).
Funding
This study was supported by National Key R& D Program of China, 2019YFD0901800, Shanghai Post-doctoral Excellence Program, 2020132, Natural Science Foundation of Shanghai, 22ZR1418200, 111 Project, B18022.
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LZ and LF conceived and supervised the project. DQ and MZ designed the experiments. DQ and CZ performed the experiments and wrote the manuscript.
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Qian, D., Zhang, C., Deng, C. et al. De novo biosynthesis of 2′-fucosyllactose in engineered Pichia pastoris. Biotechnol Lett 45, 521–536 (2023). https://doi.org/10.1007/s10529-023-03357-z
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DOI: https://doi.org/10.1007/s10529-023-03357-z