Peculiarities of the Brevundimonas diminuta Gl7ACA-acylase quaternary structure formation and obtaining stable enzyme analogues

• Published in: Applied biochemistry and microbiology Vol. 49; no. 6; pp. 549 - 554
• Main Authors: Zakirova, S. A, Mikhailova, T. V, Eldarov, M. A
• Format: Journal Article
• Language: English
• Published: Boston Springer-Verlag 01.11.2013; Springer US; Springer Nature B.V
• Subjects: Bacteria; Biochemistry; Biomedical and Life Sciences; Brevundimonas diminuta; catalytic activity; Chitin; chitinase; Enzymatic activity; enzyme activity; Enzymes; hybrids; Inactivation; Life Sciences; Medical Microbiology; Microbiology; thermal stability; Chitin; Glutaryl; Chitin Binding Domain; Apply Biochemistry; Brevundimonas
• ISSN: 0003-6838; 1608-3024
• DOI: 10.1134/S0003683813060185

The physicochemical and enzymatic properties of hybrid analogues of the Brevundimonas diminuta Gl7ACA-acylase (BrdGlA), containing the N-terminal chitin-binding domain of the bacterial chitinase (BrdGlA/NmChBD) or the C-terminal oligohistidine sequence (BrdGlA/H), were studied. An enhanced thermostability level of BrdGlA/NmChBD could suggest the stabilizing effect of the chitin-binding domain. An analysis of pH profiles of the enzymatic activity of recombinat BrdGlA analogues did not reveal significant differences: the catalytic activity of both variants changed slightly in the interval of pH values from 6.0 to 9.0 but drastically decreased at lower pH values. Both analogues demonstrated similar sensitivity towards denaturing agents: addition of 2.0 M of guanidine chloride resulted in the complete inactivation of both enzymes. A scheme was developed for obtaining isolated recombinant α- and β-subunits of BrdGLA. In vitro enzyme reconstructions indicated that the α-subunit was necessary for the formation of a correct spatial structure of the β-subunit and for the formation of a functionally active enzyme.