Recombination of thermo-alkalistable, high xylooligosaccharides producing endo-xylanase from Thermobifida fusca and expression in Pichia pastoris.

@article{Wang2015RecombinationOT,
  title={Recombination of thermo-alkalistable, high xylooligosaccharides producing endo-xylanase from Thermobifida fusca and expression in Pichia pastoris.},
  author={Qian Wang and Wen Du and X. Y. Weng and Ming-qi Liu and Jiakun Wang and Jian-xin Liu},
  journal={Applied biochemistry and biotechnology},
  year={2015},
  volume={175 3},
  pages={1318-29}
}
For xylooligosaccharide (XO) production, endo-xylanase from Thermobifida fusca was modified by error-prone PCR and DNA shuffling. The G4SM1 mutant (S62T, S144C, N198D, and A217V) showed the most improved hydrolytic activity and was two copies expressed in Pichia pastoris under the control of GAP promoter. The maximum xylanase activity in culture supernatants was 165 ± 5.5 U/ml, and the secreted protein concentration reached 493 mg/l in a 2-l baffled shake flask. After 6× His-tagged protein… CONTINUE READING

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