C-Terminal proline-rich sequence broadens the optimal temperature and pH ranges of recombinant xylanase from Geobacillus thermodenitrificans C5

IRFAN M., GÜLER H. İ., Ozer A., SAPMAZ M. T., BELDÜZ A. O., HASAN F., ...More

ENZYME AND MICROBIAL TECHNOLOGY, vol.91, pp.34-41, 2016 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 91
  • Publication Date: 2016
  • Doi Number: 10.1016/j.enzmictec.2016.05.012
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.34-41
  • Keywords: Xylanase, Geobacillus thermodinitrificans C5 recombinant xylanase (GthC5Xyl), Proline tagged xylanase (GthC5ProXyl), Characterization, Thermostability, CELLULASE-FREE, BIOCHEMICAL-CHARACTERIZATION, BACILLUS, ACID, EXPRESSION, STABILITY, CLONING, THERMOSTABILIZATION, SUBSTITUTION, ISOMERASE
  • Karadeniz Technical University Affiliated: Yes


Efficient utilization of hemicellulose entails high catalytic capacity containing xylanases. In this study, proline rich sequence was fused together with a C-terminal of xylanase gene from Geobacillus thermodenitrificans C5 and designated as GthC5ProXyl. Both GthC5Xyl and GthC5ProXyl were expressed in Escherichia coli BL21 host in order to determine effect of this modification. The C-terminal oligopeptide had noteworthy effects and instantaneously extended the optimal temperature and pH ranges and progressed the specific activity of GthC5Xyl. Compared with GthC5Xyl, GthC5ProXyl revealed improved specific activity, a higher temperature (70 degrees C versus 60 degrees C) and pH (8 versus 6) optimum, with broad ranges of temperature and pH (60-80 degrees C and 6.0-9.0 versus 40-60 degrees C and 5.0-8.0, respectively). The modified enzyme retained more than 80% activity after incubating in xylan for 3 h at 80 degrees C as compared to wild type with only 45% residual activity. Our study demonstrated that proper introduction of proline residues on C-terminal surface of xylanase family might be very effective in improvement of enzyme thermostability. Moreover, this study reveals an engineering strategy to improve the catalytic performance of enzymes. (C) 2016 Elsevier Inc. All rights reserved.