Abstract
We have previously reported that a chimeric pyrroloquinoline quinone (PQQ) glucose dehydrogenase (GDH), E97A3, which was made up of 97% of Escherichia coli PQQGDH sequence and 3% of Acinetobacter calcoaceticus PQQGDH, showed increased thermal stability compared with both parental enzymes. Site-directed mutagenesis studies were carried out in order to investigate the role of amino-acid substitution at the C-terminal region, Ser771, of a chimeric PQQGDHs on their thermal stability. A series of Ser771 substitutions of a chimeric PQQGDH, E99A1, confirmed that hydrophobic interaction governs the thermal stability of the chimeric enzymes. Comparison of the thermal denaturation of E. coli PQQGDH and E97A3 followed by far-ultraviolet (UV) circular dichroism (CD) spectroscopy revealed that E97A3 acquired stability at the first step of denaturation, which is reversible, and where no significant secondary structure change was observed. These results suggested that the interaction between C-terminal and N-terminal regions may play a crucial role in maintaining the overall structure of β-propel-ler proteins.
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© 1999 Springer Science+Business Media New York
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Witarto, A.B., Ohtera, T., Sode, K. (1999). Site-Directed Mutagenesis Study on the Thermal Stability of a Chimeric PQQ Glucose Dehydrogenase and Its Structural Interpretation. In: Davison, B.H., Finkelstein, M. (eds) Twentieth Symposium on Biotechnology for Fuels and Chemicals. Applied Biochemistry and Biotechnology. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-4612-1604-9_16
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DOI: https://doi.org/10.1007/978-1-4612-1604-9_16
Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-4612-1604-9
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