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Title (Primary) Kinetic traits and enzyme form patterns of (R)-2-(2,4-Dichlorophenoxy)propionate/a-ketoglutarate dioxygenase (RdpA) after expression in different bacterial strains
Author Westendorf, A.; Benndorf, D.; Pribyl, T.; Harms, H.; Müller, R.H.;
Journal Engineering in Life Sciences
Year 2006
Department UMB; PROTEOM;
Volume 6
Issue 6
Language englisch;
Keywords Biodegradation; Enzymes; Kinetics; Xenobiotics
Abstract The rdpA gene of strains Delftia acidovorans MC1, Rhodoferax sp. P230, and Sphingobium herbicidovorans MH proved to be identical. However, when RdpA [(R)-2-(2,4-dichlorophenoxy)propionate/-ketoglutarate dioxygenase] was investigated after purification from the various strains, significant differences in the kinetics and some chemical properties of the enzymes were observed. The preference for substrates ranged in the order (R)-2-(2,4-dichlorophenoxy)propionate (2,4-DP) > (R)-2-(4-chloro-2-methylphenoxy)propionate (MCPP) >> 2,4-dichlorophenoxyacetate (2,4-D) 4-chloro-2-methylphenoxyacetate (MCPA), but detailed kinetic investigations revealed significant strain-dependent differences in the kcat and KM values. While the KM values of RdpA from the various strains were low and their range rather narrow with 2,4-DP (19-60 m M) and MCPP (35-64 m M), larger differences were observed with phenoxyacetates which were distinctly higher and spanned a wider range with 2,4-D (237-935 m M) and MCPA (164-510 m M). The lowest KM values with 2,4-D and MCPA were found for RdpA originating from strain P230. Investigation of the enzymes from the various sources by 2D gel electrophoresis revealed up to three monomeric enzyme forms which differed in the pI value. The 2D-patterns were similar with RdpA from strains MC1 and MH, and after heterologous expression of the enzyme in Escherichia coli, but differed significantly from that of strain P230. The presence of enzyme forms and their different composition coincided apparently with the differences observed in the kinetic properties of RdpA in the various strains. The effects are discussed in terms of posttranslational modification of RdpA which appears to be different in extent and kind in the various strains.
ID 3121
Persistent UFZ Identifier https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=3121
Westendorf, A., Benndorf, D., Pribyl, T., Harms, H., Müller, R.H. (2006):
Kinetic traits and enzyme form patterns of (R)-2-(2,4-Dichlorophenoxy)propionate/a-ketoglutarate dioxygenase (RdpA) after expression in different bacterial strains
Eng. Life Sci. 6 (6), 552 - 559