Chemoenzymatic syntheses of water-soluble lipid I fluorescent probes

• Published in: Tetrahedron letters Vol. 56; no. 23; pp. 3441 - 3446
• Main Authors: Mitachi, Katsuhiko, Siricilla, Shajila, Klaić, Lada, Clemons, William M., Kurosu, Michio
• Format: Journal Article
• Language: English
• Published: OXFORD Elsevier Ltd 03.06.2015; Elsevier
• Subjects: Chemistry; Chemistry, Organic; Chemoenzymatic synthesis; Lipid I; Lipid II; MurG; Physical Sciences; Science & Technology; Translocase I (MraY/MurX); MurG; Lipid II; Chemoenzymatic synthesis; Translocase I (MraY/MurX); Lipid I; PRECURSOR; PARKS NUCLEOTIDE; MYCOBACTERIA; BIOSYNTHESIS; MRAY; INHIBITORS; PEPTIDOGLYCAN SYNTHESIS; CAPURAMYCIN; DERIVATIVES; BACTERIAL-CELL WALL
• ISSN: 0040-4039; 1873-3581
• DOI: 10.1016/j.tetlet.2015.01.044

[Display omitted] Peptidoglycan (PG) is unique to bacteria, and thus, the enzymes responsible for its biosynthesis are promising antibacterial drug targets. The membrane-embedded enzymes in PG remain significant challenges in studying their mechanisms due to the fact that preparations of suitable enzymatic substrates require time-consuming biological transformations or chemical synthesis. Lipid I (MurNAc(pentapeptide)-pyrophosphoryl prenol) is an important PG biosynthesis intermediate to study the central enzymes, translocase I (MraY/MurX) and MurG. Lipid I isolated from nature contains the C50- or C55-prenyl unit that shows extremely poor water-solubility that renders studies of translocase I and MurG enzymes difficult. We have studied biological transformation of water soluble lipid I fluorescent probes using bacterial membrane fractions and purified MraY enzymes. In our investigation of the minimum structural requirements of the prenyl phosphates in MraY-catalyzed lipid I synthesis, we found that (2Z,6E)-farnesyl phosphate (C15-phosphate) can be recognized by Escherichia coli MraY to generate the water-soluble lipid I fluorescent probe in high-yields. Under the optimized conditions, the same reaction was performed by using the purified MraY from Hydrogenivirga spp. to afford the lipid I analog with high-yields in a short reaction time.