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Biochemical characterization of a novel thermostable feruloyl esterase from Geobacillus thermoglucosidasius DSM 2542T

  • Fulya Ay Sal
  • Dilsat Nigar Colak
  • Halil Ibrahim Guler
  • Sabriye Canakci
  • Ali Osman BelduzEmail author
Original Article
  • 35 Downloads

Abstract

The ferulic acid esterase (FAE) gene from Geobacillus thermoglucosidasius DSM 2542T was cloned into pET28a(+) expression vector and characterized and is being reported in this study for the first time in Geobacillus. The enzyme, designated as GthFAE, was purified by heat shock and ion-exchange column chromatography. In addition, a second clone containing a Histidine tag was expressed and purified by affinity column chromatography demonstrating future potential for scale-up. FAE gene contains an open reading frame (ORF) of 759-bp encoding a hypothetical 252 amino acid protein, a molecular mass of 28.11 kDa and an isoelectric point of 5.53. From this study it was found that GthFAE had optimal activity at 50 °C and pH of 8.5. Furthermore, the enzyme has been found to retain 64% of its activity after two days incubation at 50 °C and exhibited a high level of functionality with p-nitrophenyl caprylate (C8). Km, Vmax, kcat and kcat/Km values for p-nitrophenyl caprylate were determined as 0.035 mM, 11,735 µmol/min/mg protein, 5491 (1/s) and 156,885 s−1 mM−1 respectively. The combination of higher activity and stability compared to previously reported FAEs makes GthFAE a potential candidate for use in the paper manufacturing industry.

Keywords

Geobacillus Feruloyl esterase Recombinant Thermostable 

Notes

Acknowledgements

We would like to thank Dr. Asif Rahman (Space Biosciences Division, NASA) for critical reading and language editing of this manuscript. This work was supported by the Karadeniz Technical University Research Foundation [Grant No. BAP01-8704].

Supplementary material

11033_2019_4893_MOESM1_ESM.pdf (479 kb)
Supplementary material 1 (PDF 479 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Fulya Ay Sal
    • 1
  • Dilsat Nigar Colak
    • 2
  • Halil Ibrahim Guler
    • 3
  • Sabriye Canakci
    • 1
  • Ali Osman Belduz
    • 1
    Email author
  1. 1.Department of Biology, Faculty of SciencesKaradeniz Technical UniversityTrabzonTurkey
  2. 2.Department of Forestry, Vocational School of DereliGiresun UniversityGiresunTurkey
  3. 3.Department of Molecular Biology and Genetics, Faculty of SciencesKaradeniz Technical UniversityTrabzonTurkey

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