Functional characterization of a thermostable methionine adenosyltransferase from Thermus thermophilus HB27

Abstract

MATTt (a thermostable methionine adenosyltransferase from Thermus thermophilus HB27) was overexpressed in Escherchia coli and purified using Ni-NTA affinity column. The enzymatic activity of MATTt was investigated in a temperature range from 30 °C to 90 °C, showing that MATTt exhibited a high enzymatic activity and good thermostability at 80 °C. Circular dichroism spectra reveals that MATTt contains high portion of β-sheet structures contributing to the thermostability of MATTt. The kinetic parameter, K_m is 4.19 mmol/L and 1.2 mmol/L for ATP and methionine, respectively. MATTt exhibits the highest enzymatic activity at pH 8. Cobalt (Co²⁺) and zinc ion (Zn²⁺) enhances remarkably the activity of MATTt compared to the magnesium ion (Mg²⁺). All these results indicated that the thermostable MATTt has great potential for industry applications.

Graphical abstract

Keywords

ion-preference / methionine adenosyltransferase / secondary structure / thermostability / Thermus thermophilus

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Acknowledgement

This work was financially supported by the National 973 Basic Research Program of China (2014CB745100), National Natural Science Foundation of China (Grant Nos. 21376024 and 21390202).

Electronic Supplementary Material

ƒSupplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s11705-016-1566-2 and is accessible for authorized users.

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