<![CDATA[Developing semisynthetic penicillin is one approach to overcoming antibiotic resistance. Isopenicillin N synthase (IPNS) is crucial for the biosynthesis of semisynthetic penicillin, as it catalyzes the oxidative condensation of delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (LLD-ACV) to isopenicillin N. Through genome mining studies of Serratia plymuthica strain UBCF_13, the presence of the pcbC gene encoding isopenicillin N synthase family oxygenase (IPNSfo) was revealed. Therefore, this study aimed to clone the pcbC gene from S. plymuthica UBCF_13 into Escherichia coli and to characterize it at the DNA and in silico level. To achieve this, the pcbC gene was first amplified using PCR with specific primers, producing a single amplicon of approximately 1035 bp, and then cloned into the pGEM-T Easy vector. Colony PCR verified the presence of the recombinant pGEM_pcbC construct in transformant bacteria. The cloned insert was subsequently sequenced, and the edited consensus sequence showed 100% similarity and 0% phylogenetic distance to the S. plymuthica UBCF_13 genome. For bioinformatics characterization, the predicted IPNSfo protein structure showed consistent features, and domain analysis identified three conserved domains: DIOX_N, IPNS-like_FE2OG_OXY, and Oxoglu/Fe_dep_dioxygenase. These findings demonstrate the successful cloning and preliminary molecular characterization of the pcbC gene from S. plymuthica UBCF_13 in E. coli.]]>
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