Qian Difan, Zhang Chunyue, Deng Chen, Zhou Mian, Fan Liqiang, Zhao Liming
State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Meilong Road 130, Shanghai, 200237, China.
Shanghai Collaborative Innovation Center for Biomanufacturing Technology (SCICBT), Shanghai, 200237, China.
Biotechnol Lett. 2023 Apr;45(4):521-536. doi: 10.1007/s10529-023-03357-z. Epub 2023 Feb 15.
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).
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 P promoter.
The resulting recombinant yeasts yielded up to 0.276 g/L through culture optimization in a 5 L bioreactor.
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.
巴斯德毕赤酵母以其在重组糖蛋白上产生短且低免疫原性的人源化糖链的能力而闻名，因此推测它可用于合成低聚糖。在本研究中，对一般认为安全（GRAS）的微生物巴斯德毕赤酵母GS115作为一种新的合成底盘来生产最丰富的人乳低聚糖2'-岩藻糖基乳糖（2'-FL）的潜力进行了测试。
为了实现2'-FL的从头合成，将乳糖转运蛋白lac12、两种酶gmd、gmer和岩藻糖基转移酶futC在组成型P启动子的控制下整合到巴斯德毕赤酵母的基因组中。
通过在5 L生物反应器中进行培养优化，所得重组酵母的产量高达0.276 g/L。
据我们所知，这是关于工程化巴斯德毕赤酵母生产2'-FL的首次报道。这项工作是以巴斯德毕赤酵母作为可行底盘生产2'-FL的良好起点。
