The specificity of feruloyl esterase from Geobacillus thermoglucosidasius towards model substrates by molecular docking

Ay Şal F., Karaoğlu H., Beldüz A. O.

FEMS Conference on Microbiology, Belgrade, Sırbistan, 30 Haziran - 02 Temmuz 2022, ss.113

  • Yayın Türü: Bildiri / Özet Bildiri
  • Basıldığı Şehir: Belgrade
  • Basıldığı Ülke: Sırbistan
  • Sayfa Sayıları: ss.113
  • Karadeniz Teknik Üniversitesi Adresli: Evet

Özet

Feruloyl esterases (FAEs) are hydrolase enzymes, (EC 3.1.1.73) a subclass of carboxylic acid esterases (EC 3.1.1), that catalyze the formation and breakdown of ester bonds between the ferulate ester groups required for crosslinking between hemicelluloses and hemicellulose-lignin.

The aim of this study is to investigate the affinity of FAE for four different model substrates (methyl/ethyl ferulat, methyl caffeine, methyl-p-coumarate, and methyl sinapat) and to support these datas by molecular docking. For this, FAE from Geobacillus thermoglucosidasius DSM 2542T which was cloned into pET28a(+) and expressed in Escherichia coli was used. Expressed enzyme was purified by heat shock and anionic ion exchange column chromatography. Four model substrates of FAE were used and determined which substrate the enzyme has more affinity for. Molecular docking studies were performed using AutoDock 4.2 software to investigate possible interactions of methyl and arabinose forms of model substrates. In this study, eight different compounds were docked to the 3D structure of feruloyl esterase from Geobacillus thermoglucosidasius DSM 2542T.

The catalytic domain of the enzyme was determined as Ser114-His232-Asp202, taking into account the literature studies in previous studies. In the docking studies performed according to this region, it was observed that all substrates could bind to this region. According to these results, the substrate with the lowest binding energy (the substrate that forms the most stable complex with the enzyme) was determined as arabinose ferulate (-7.32 kcal/mol).