Engineering Escherichia coli for constitutive production of monophosphoryl lipid A vaccine adjuvant

• Published in: Biotechnology and bioengineering 2024-03, Vol.121 (3), p.1144-1162
• Main Authors: Jin, Hyunjung, Ji, Yuhyun, An, Jinsu, Ha, Da Hui, Lee, Ye‐Ram, Kim, Hye‐Ji, Lee, Choon Geun, Jeong, Wooyeon, Kwon, Ick Chan, Yang, Eun Gyeong, Kim, Ki Hun, Lee, Chankyu, Chung, Hak Suk
• Format: Article
• Language: English
• Subjects: Additives; adjuvant; adjuvant production; Adjuvants; Antibiotics; Commercialization; COVID-19; COVID-19 vaccines; E coli; Effectiveness; Escherichia coli; Fermentation; Herpes zoster; Lipid A; lipid A 1‐phosphatase; Lipids; lipopolysaccharide biosynthetic pathway; metabolic engineering; Monophosphoryl lipid A; Pandemics; Peptidoglycans; Production costs; Production methods; Public health; Raw materials; Vaccines
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.28638

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. Building on the fact that LpxE, a bifunctional lipid A 1‐phosphatase, can functionally rescue the lethality of E. coli lacking C55‐PP phosphatase activity, Jin et al. undertook metabolic engineering to create an E. coli cell factory that constitutively produces EcML (E. coli‐produced monophosphoryl lipid A), a valuable vaccine adjuvant. This innovative approach allows for the production of EcML without the need for antibiotics or overexpression inducers, paving the way for cost‐effective, industrial‐grade EcML adjuvant production for human use.