Complete biosynthesis of QS-21 in engineered yeast

• 成果类型: Article
• 署名作者: Liu, Yuzhong; Zhao, Xixi; Gan, Fei; Chen, Xiaoyue; Deng, Kai; Crowe, Samantha A.; Hudson, Graham A.; Belcher, Michael S.; Schmidt, Matthias; Astolfi, Maria C. T.; Kosina, Suzanne M.; Pang, Bo; Shao, Minglong; Yin, Jing; Sirirungruang, Sasilada; Iavarone, Anthony T.; Reed, James; Martin, Laetitia B. B.; El-Demerdash, Amr; Kikuchi, Shingo; Misra, Rajesh Chandra; Liang, Xiaomeng; Cronce, Michael J.; Chen, Xiulai; Zhan, Chunjun; Kakumanu, Ramu; Baidoo, Edward E. K.; Chen, Yan; Petzold, Christopher J.; Northen, Trent R.; Osbourn, Anne; Scheller, Henrik; Keasling, Jay D.
• 署名单位: University of California System; University of California Berkeley; United States Department of Energy (DOE); Joint BioEnergy Institute - JBEI; University of California System; University of California Berkeley; United States Department of Energy (DOE); Lawrence Berkeley National Laboratory; United States Department of Energy (DOE); Sandia National Laboratories; University of California System; University of California Berkeley; United States Department of Energy (DOE); Lawrence Berkeley National Laboratory; University of California System; University of California Berkeley; Suranaree University of Technology; UK Research & Innovation (UKRI); Biotechnology and Biological Sciences Research Council (BBSRC); John Innes Center; Egyptian Knowledge Bank (EKB); Mansoura University; Technical University of Denmark
• 刊物名称: Nature
• ISSN/ISSBN: 0028-5481
• DOI: 10.1038/s41586-024-07345-9
• 发表日期: 2024-05-23
• 页码: 937-+
• 关键词: vaccine adjuvants; cytochrome b(5); udp-glucose; expression; proteins

摘要

QS-21 is a potent vaccine adjuvant and remains the only saponin-based adjuvant that has been clinically approved for use in humans(1,2). However, owing to the complex structure of QS-21, its availability is limited. Today, the supply depends on laborious extraction from the Chilean soapbark tree or on low-yielding total chemical synthesis(3,4). Here we demonstrate the complete biosynthesis of QS-21 and its precursors, as well as structural derivatives, in engineered yeast strains. The successful biosynthesis in yeast requires fine-tuning of the host's native pathway fluxes, as well as the functional and balanced expression of 38 heterologous enzymes. The required biosynthetic pathway spans seven enzyme families-a terpene synthase, P450s, nucleotide sugar synthases, glycosyltransferases, a coenzyme A ligase, acyl transferases and polyketide synthases-from six organisms, and mimics in yeast the subcellular compartmentalization of plants from the endoplasmic reticulum membrane to the cytosol. Finally, by taking advantage of the promiscuity of certain pathway enzymes, we produced structural analogues of QS-21 using this biosynthetic platform. This microbial production scheme will allow for the future establishment of a structure-activity relationship, and will thus enable the rational design of potent vaccine adjuvants. QS-21-an FDA-approved vaccine adjuvant-and several structural analogues of QS-21 can be synthesized in engineered yeast strains, and this process is much less laborious compared with the conventional mode of extraction from the Chilean soapbark tree.