Ji, Yuhyun An, Jinsu Hwang, Dohyeon Ha, Da Hui Lim, Sang Min Lee, Chankyu Zhao, Jinshi Song, Hyun Kyu Yang, Eun Gyeong Zhou, Pei Chung, Hak Suk 2024-01-19T18:31:44Z 2024-01-19T18:31:44Z 2021-09-05 2020-01 1096-7176 https://pubs.kist.re.kr/handle/201004/119150 Monophosphoryl lipid A (MPLA) species, including MPL (a trade name of GlaxoSmithKline) and GLA (a trade name of Immune Design, a subsidiary of Merck), are widely used as an adjuvant in vaccines, allergy drugs, and immunotherapy to boost the immune response. Even though MPLA is a derivative of lipopolysaccharide (LPS), a component of the outer membrane of Gram-negative bacteria, bacterial strains producing MPLA have not been found in nature nor engineered. In fact, MPLA generation involves expensive and laborious procedures based on synthetic routes or chemical transformation of precursors isolated from Gram-negative bacteria. Here, we report the engineering of an Escherichia coli strain for in situ production and accumulation of MPLA. Furthermore, we establish a succinct method for purifying MPLA from the engineered E. coli strain. We show that the purified MPLA (named EcML) stimulates the mouse immune system to generate antigen-specific IgG antibodies similarly to commercially available MPLA, but with a dramatically reduced manufacturing time and cost. Our system, employing the first engineered E. coli strain that directly produces the adjuvant EcML, could transform the current standard of industrial MPLA production. English ACADEMIC PRESS INC ELSEVIER SCIENCE Metabolic engineering of Escherichia coli to produce a monophosphoryl lipid A adjuvant Article 10.1016/j.ymben.2019.11.009 1 METABOLIC ENGINEERING, v.57, pp.193 - 202 METABOLIC ENGINEERING 57 193 202 N scie scopus 000506206200019 2-s2.0-85076692796 Biotechnology & Applied Microbiology Biotechnology & Applied Microbiology Article EFFECTIVE VACCINE LIPOPOLYSACCHARIDE 1-PHOSPHATASE PURIFICATION EXPRESSION MEMBRANE CLONING MUTANT LPXE Adjuvant Monophosphoryl lipid A Lipopolysaccharide biosynthesis Gram-negative bacterial outer membrane Lipid A 1-phosphatase Vaccine adjuvant