Molecular Identification and Engineering a Salt-Tolerant GH11 Xylanase for Efficient Xylooligosaccharides Production

This study identified a salt-tolerant GH11 xylanase, Xyn , which was isolated from a soil bacterium sp. SC1 and can resist as high as 4 M NaCl. After rational design and high-throughput screening of site-directed mutant libraries, a double mutant W6F/Q7H with a 244% increase in catalytic activity an...

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Veröffentlicht in:Biomolecules (Basel, Switzerland) Switzerland), 2024-09, Vol.14 (9), p.1188
Hauptverfasser: Ma, Jiao, Sun, Zhongke, Ni, Zifu, Qi, Yanli, Sun, Qianhui, Hu, Yuansen, Li, Chengwei
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Sprache:eng
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Zusammenfassung:This study identified a salt-tolerant GH11 xylanase, Xyn , which was isolated from a soil bacterium sp. SC1 and can resist as high as 4 M NaCl. After rational design and high-throughput screening of site-directed mutant libraries, a double mutant W6F/Q7H with a 244% increase in catalytic activity and a 10 °C increment in optimal temperature was obtained. Both Xyn and W6F/Q7H xylanases were stimulated by high concentrations of salts. In particular, the activity of W6F/Q7H was more than eight times that of Xyn in the presence of 2 M NaCl at 65 °C. Kinetic parameters indicated they have the highest affinity for beechwood xylan ( = 0.30 mg mL for Xyn and 0.18 mg mL for W6F/Q7H), and W6F/Q7H has very high catalytic efficiency ( / = 15483.33 mL mg s ). Molecular dynamic simulation suggested that W6F/Q7H has a more compact overall structure, improved rigidity of the active pocket edge, and a flexible upper-end alpha helix. Hydrolysis of different xylans by W6F/Q7H released more xylooligosaccharides and yielded higher proportions of xylobiose and xylotriose than Xyn did. The conversion efficiencies of Xyn and W6F/Q7H on all tested xylans exceeded 20%, suggesting potential applications in the agricultural and food industries.
ISSN:2218-273X
2218-273X
DOI:10.3390/biom14091188