Deciphering the thermotolerance of chitinase O from Chitiniphilus shinanonensis by in vitro and in silico studies
Biochemical and biophysical studies revealed that chitinase O from Chitiniphilus shinanonensis (CsChiO) exhibits considerable thermotolerance, possibly due to the formation of a stable structural conformation. CsChiO is an exochitinase with a temperature optimum of 70 °C. The secondary structures of...
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Veröffentlicht in: | International journal of biological macromolecules 2022-06, Vol.210, p.44-52 |
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Sprache: | eng |
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Zusammenfassung: | Biochemical and biophysical studies revealed that chitinase O from Chitiniphilus shinanonensis (CsChiO) exhibits considerable thermotolerance, possibly due to the formation of a stable structural conformation. CsChiO is an exochitinase with a temperature optimum of 70 °C. The secondary structures of CsChiO and its catalytic domain (Cat-CsChiO) are only marginally affected upon heating up to 90 °C, as revealed by circular dichroism (CD) spectroscopy. Differential scanning calorimetric (DSC) studies revealed that CsChiO exhibits two endothermic transitions at ca. 51 °C (Tm1) and 59 °C (Tm2), whereas Cat-CsChiO shows a single endothermic transition at 52 °C. Together, the CD and DSC analyses suggested that the catalytic domain of CsChiO undergoes a thermotropic transition at ~52 °C from native state to another stable structural conformation. Results from molecular dynamic simulations corroborated that Cat-CsChiO adopts a stable structural conformation above 50 °C by partial unfolding. Thermotolerant CsChiO would be useful for the conversion of chitin, which is highly abundant, to biologically active COS. This study unveiled the adaptability of enzymes/proteins in nature to perform biological functions at elevated temperatures.
•CsChiO is a thermotolerant exochitinase with optimal activity at 70 °C.•Secondary structures of CsChiO and Cat-CsChiO are mostly unaltered up to 90 °C.•Catalytic domain of CsChiO adopts a molten globule-like structure above 50 °C.•Binding of chitooligosaccharides increases the thermal stability of CsChiO. |
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ISSN: | 0141-8130 1879-0003 |
DOI: | 10.1016/j.ijbiomac.2022.05.013 |