Recombinant Production of E. coli NAD(+)-dependent DNA ligase as a Target for Antibacterial Drug Discovery

KAPLAN Ö., Imamoglu R., Gokce I.

KSU TARIM VE DOGA DERGISI-KSU JOURNAL OF AGRICULTURE AND NATURE, cilt.25, sa.1, ss.19-24, 2022 (ESCI) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 25 Sayı: 1
  • Basım Tarihi: 2022
  • Doi Numarası: 10.18016/ksutarimdoga.vi.884279
  • Dergi Adı: KSU TARIM VE DOGA DERGISI-KSU JOURNAL OF AGRICULTURE AND NATURE
  • Derginin Tarandığı İndeksler: Emerging Sources Citation Index (ESCI), TR DİZİN (ULAKBİM)
  • Sayfa Sayıları: ss.19-24
  • Anahtar Kelimeler: NAD(+)-dependent DNA ligase, TolAIII peptide, Antibacterial Drug Discovery, ESCHERICHIA-COLI, CRYSTAL-STRUCTURE, EXPRESSION, CLONING, IDENTIFICATION, SEQUENCE, BINDING, DOMAIN
  • İstanbul Üniversitesi Adresli: Evet

Özet

The increase in the frequency of drug resistance in bacterial infections has led to the research of new antibacterial agents targeting new have made it a remarkable target for antibacterial drug discovery. a potential target due to its unique substrate specificity compared to the ATP-dependent human DNA ligase. In this study, it was aimed to produce and purify the E. coli NAD(+)-dependent DNA ligase enzyme, which is frequently used in antibacterial drug discovery. The E. coli DNA ligase gene sequence was cloned into pTOLT vector system. E. coli DNA ligase enzyme was purified after the production in E. coli BL21 (DE3) pLysE cells. It was clearly demonstrated by the activity test that the DNA ligase enzyme produced in this study can ligate the DNA fragments. As a result, it was revealed that the effect of candidate inhibitors can be studied simply on the enzyme.