5FLG

Crystal structure of the 6-carboxyhexanoate-CoA ligase (BioW)from Bacillus subtilis in complex with AMPPNP

Summary for 5FLG

• Entry DOI: 10.2210/pdb5flg/pdb
• Related: 5FLL 5FM0
• Descriptor: 6-CARBOXYHEXANOATE--COA LIGASE, PIMELIC ACID, PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER, ... (5 entities in total)
• Functional Keywords: ligase
• Biological source: BACILLUS SUBTILIS
• Total number of polymer chains: 2
• Total formula weight: 60778.79

Authors

Moynie, L.,Wang, M.,Campopiano, D.J.,Naismith, J.H. (deposition date: 2015-10-26, release date: 2016-11-16, Last modification date: 2024-05-08)

Primary citation

Wang, M.,Moynie, L.,Harrison, P.J.,Kelly, V.,Piper, A.,Naismith, J.H.,Campopiano, D.J.
Using the pimeloyl-CoA synthetase adenylation fold to synthesize fatty acid thioesters.
Nat. Chem. Biol., 13:660-667, 2017

PubMed Abstract: Biotin is an essential vitamin in plants and mammals, functioning as the carbon dioxide carrier within central lipid metabolism. Bacterial pimeloyl-CoA synthetase (BioW) acts as a highly specific substrate-selection gate, ensuring the integrity of the carbon chain in biotin synthesis. BioW catalyzes the condensation of pimelic acid (C7 dicarboxylic acid) with CoASH in an ATP-dependent manner to form pimeloyl-CoA, the first dedicated biotin building block. Multiple structures of Bacillus subtilis BioW together capture all three substrates, as well as the intermediate pimeloyl-adenylate and product pyrophosphate (PP), indicating that the enzyme uses an internal ruler to select the correct dicarboxylic acid substrate. Both the catalytic mechanism and the surprising stability of the adenylate intermediate were rationalized through site-directed mutagenesis. Building on this understanding, BioW was engineered to synthesize high-value heptanoyl (C7) and octanoyl (C8) monocarboxylic acid-CoA and C8 dicarboxylic-CoA products, highlighting the enzyme's synthetic potential.

PubMed: 28414710
DOI: 10.1038/nchembio.2361

Experimental method

X-RAY DIFFRACTION (2.04 Å)