Kinetics of Thermoinactivation of D-Amino Acid Oxidase OPADAAO1 from the Ogataea parapolymorpha DL-1 Yeast

• Published in: Moscow University chemistry bulletin Vol. 78; no. 2; pp. 69 - 75
• Main Authors: Koshkina, M. K., Sergeyev, E. P., Fedorov, T. A., Shelomov, M. D., Pometun, A. A., Savin, S. S., Tishkov, V. I., Atroshenko, D. L.
• Format: Journal Article
• Language: English
• Published: Moscow Pleiades Publishing 01-04-2023; Springer Nature B.V
• Subjects: Amino acid oxidase; Amino acids; Annotations; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; Deactivation; E coli; Enzymes; Genes; High temperature; Kinetics; Original Article; Thermal stability; Yeast; thermal inactivation kinetics; temperature stability; DL-1; D-amino acid oxidase
• ISSN: 0027-1314; 1935-0260
• DOI: 10.3103/S0027131423020049

Our earlier annotation of the genome of the yeast
Ogataea parapolymorpha
DL-1 made it possible to identify five genes of potential D-amino acids oxidases. All
opadaao1
–
opadaao5
genes were cloned and expressed in
E. coli
. Four OpaDAAO1-OpaDAAO4 enzymes were obtained in highly purified form and their catalytic properties were studied. It was found that among all DAAO described in the literature, the enzyme OpaDAAOl has the highest catalytic constant
k
cat
with D-Ala, which makes it promising for practical applications. However, in addition to good catalytic parameters, effective application of the enzyme in practice requires stability and knowledge of the inactivation mechanism, including at elevated temperatures. In this paper, we study the effect of elevated temperatures on the stability of OpaDAAOl. The enzyme is shown to have higher thermal stability than the majority of other D-amino acid oxidases. The kinetics of OpaDAAOl inactivation at different temperatures, at the initial concentrations of the enzyme, and in the presence of exogenous FAD are studied. A possible kinetic scheme of inactivation is proposed based on the data obtained.