Hyunjung Jin Yuhyun Ji Jinsu An Da Hui Ha Ye­-Ram Lee Hye­-Ji Kim Choon Geun Lee Wooyeon Jeong 권익찬 양은경 김기훈 Chankyu Lee 정학숙 2024-01-12T06:30:32Z 2024-01-12T06:30:32Z 2024-01-08 2024-03 0006-3592 https://pubs.kist.re.kr/handle/201004/79671 During the COVID-19 pandemic, expedient vaccine production has been slowed by the shortage of safe and effective raw materials, such as adjuvants, essential components to enhance the efficacy of vaccines. Monophosphoryl lipid A (MPLA) is a potent and safe adjuvant used in human vaccines, including the Shingles vaccine, Shingrix. 3-O-desacyl-4′-monophosphoryl lipid A (MPL), a representative MPLA adjuvant commercialized by GSK, was prepared via chemical conversion of precursors isolated from Salmonella typhimurium R595. However, the high price of these materials limits their use in premium vaccines. To combat the scarcity and high cost of safe raw materials for vaccines, we need to develop a feasible MPLA production method that is easily scaled up to meet industrial requirements. In this study, we engineered peptidoglycan and outer membrane biosynthetic pathways in Escherichia coli and developed a Escherichia coli strain, KHSC0055, that constitutively produces EcML (E. coli-produced monophosphoryl lipid A) without additives such as antibiotics or overexpression inducers. EcML production was optimized on an industrial scale via high-density fed-batch fermentation, and obtained 2.7？g of EcML (about 135,000 doses of vaccine) from a 30-L-scale fermentation. Using KHSC0055, we simplified the production process and decreased the production costs of MPLA. Then, we applied EcML purified from KHSC0055 as an adjuvant for a COVID-19 vaccine candidate (EuCorVac-19) currently in clinical trial stage III in the Philippines. By probing the efficacy and safety of EcML in humans, we established KHSC0055 as an efficient cell factory for MPLA adjuvant production. English Wiley - V C H Verlag GmbbH & Co. Engineering Escherichia coli for constitutive production of monophosphoryl lipid A vaccine adjuvant Article 10.1002/bit.28638 1 Biotechnology and Bioengineering, v.121, no.3, pp.1144 - 1162 Biotechnology and Bioengineering 121 3 1144 1162 N scie scopus 001137237500001 Biotechnology & Applied Microbiology Biotechnology & Applied Microbiology Article EXPRESSION PROTEINS VECTORS SYSTEM GENES lipid A 1-phosphatase lipopolysaccharide biosynthetic pathway metabolic engineering monophosphoryl lipid A adjuvant adjuvant production