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Lisana Shiddiqin Aliyya
Department of Biomedical Sciences, Medical Faculty, Universitas Andalas, Padang, Indonesia
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Materials Science & Nanotechnology Medicine & Pharmacology

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Cloning of the pcbC Gene Encoding Isopenicillin N Synthase Family Oxygenase from Serratia plymuthica UBCF_13 in Escherichia coli Fauzan Syarif Nursyafi; Abi Awfa Rahman; Imron Hasibuan; Lisana Shiddiqin Aliyya; Noza Hilbertina; Dian Pertiwi; Hasmiwati Hasmiwati; Julizar Nazar; Lily Syukriani; Jamsari Jamsari
Biota Vol 12 No 2 (2026): Jurnal Biota
Publisher : Faculty of Science and Technology Universitas Islam Negeri Raden Fatah Palembang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.19109/biota.v12i2.30398

Abstract

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.
Co-Authors Abi Awfa Rahman Dian Pertiwi Fauzan Syarif Nursyafi Hasmiwati Hasmiwati Imron Hasibuan Jamsari Jamsari Julizar Nazar Lily Syukriani Noza Hilbertina