Products/Services Used | Details | Operation |
---|---|---|
Gene Synthesis> | Cloning, expression and purification. The full-length, codon-optimized bioW gene (Bacillus subtilis (strain 168), UNIPROT code: P53559) was purchased from GenScript and cloned into two expression plasmids to give recombinant BioW with a noncleavable N-terminal His6 tag (pET-28a) and a pEHISTEV plasmid, which gives an N-terminal tobacco etch virus (TEV) protease-cleavable His6 tag43 . | Get A Quote |
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 (PPi ), indicating that the enzyme uses an internal ruler to select the correct dicarboxylic acid subs... More