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Thermal behaviour and tolerance to ionic liquid [emim]OAc in GH10 xylanase from Thermoascus aurantiacus SL16W

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Abstract

GH10 xylanase from Thermoascus aurantiacus strain SL16W (TasXyn10A) showed high stability and activity up to 70–75 °C. The enzyme’s half-lives were 101 h, 65 h, 63 min and 6 min at 60, 70, 75 and 80 °C, respectively. The melting point (T m), as measured by DSC, was 78.5 °C, which is in line with a strong activity decrease at 75–80 °C. The biomass-dissolving ionic liquid 1-ethyl-3-methylimidazolium acetate ([emim]OAc) in 30 % concentration had a small effect on the stability of TasXyn10A; T m decreased by only 5 °C. It was also observed that [emim]OAc inhibited much less GH10 xylanase (TasXyn10A) than the studied GH11 xylanases. The K m of TasXyn10A increased 3.5-fold in 15 % [emim]OAc with xylan as the substrate, whereas the approximate level of V max was not altered. The inhibition of enzyme activity by [emim]OAc was lesser at higher substrate concentrations. Therefore, high solid concentrations in industrial conditions may mitigate the inhibition of enzyme activity by ionic liquids. Molecular docking experiments indicated that the [emim] cation has major binding sites near the catalytic residues but in lower amounts in GH10 than in GH11 xylanases. Therefore, [emim] cation likely competes with the substrate when binding to the active site. The docking results indicated why the effect is lower in GH10.

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Acknowledgments

This work was financially supported by the Thailand Research Fund through the Royal Golden Jubilee Ph.D. grant, Contract No. PHD/0189/2548 and Centre of Excellent for Renewable Energy, Chiang Mai University. We thank Johanna Aura for technical assistance.

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Authors and Affiliations

  1. Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand

    Niwat Chawachart & Saisamorn Lumyong

  2. Department of Biotechnology and Chemical Technology, School of Chemical Technology, Aalto University, P.O. Box 16100, 00076, Aalto, Finland

    Sasikala Anbarasan, Samuel Turunen, He Li, Tom Granström & Ossi Turunen

  3. Department of Biotechnology, Faculty of Agro-Industry, Chiang Mai University, Chiang Mai, 50200, Thailand

    Chartchai Khanongnuch

  4. Department of Forest Products Technology, School of Chemical Technology, Aalto University, P.O. Box 16300, 00076, Aalto, Finland

    Michael Hummel & Herbert Sixta

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  1. Niwat Chawachart
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  2. Sasikala Anbarasan
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  3. Samuel Turunen
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  4. He Li
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  9. Saisamorn Lumyong
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  10. Ossi Turunen
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Corresponding author

Correspondence to Ossi Turunen.

Additional information

Communicated by L. Huang.

N. Chawachart and S. Anbarasan contributed equally to the study.

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Chawachart, N., Anbarasan, S., Turunen, S. et al. Thermal behaviour and tolerance to ionic liquid [emim]OAc in GH10 xylanase from Thermoascus aurantiacus SL16W. Extremophiles 18, 1023–1034 (2014). https://doi.org/10.1007/s00792-014-0679-0

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