Evolution of enzymatic activities: Multiple pathways for generating and partitioning a common enolic intermediate by glucarate dehydratase from Pseudomonas putida

Journal of the American Chemical Society

Volumn 118, Issue 42, 1996, Pages 10323-10324

  Palmer, David R J ^a   Gerlt, John A ^a  

^a UNIVERSITY OF ILLINOIS AT URBANA CHAMPAIGN   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

HYDROLYASE;

ARTICLE; ENZYME ACTIVITY; PSEUDOMONAS PUTIDA;

EID: 0029963388     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja962126v     Document Type: Article

References (14)

1. Petsko, G. A.; Kenyon, G. L.; Gerlt, J. A.; Ringe, D. A.; Kozarich, J. W. Trends Biochem. Sci. 1993, 18, 372.
2. Babbitt, P. C; Hasson, M.; Wedekind, J. E.; Palmer, D. R. J.; Barrett, W. C.; Rayment, I.; Ringe, D.; Kenyon, G. L.; Gerlt, J. A. Biochemistry, in press.
3. Kenyon, G. L.; Gerlt, J. A.; Petsko, G. A.; Kozarich, J. W. Acc. Chem. Res. 1995, 28, 178.
4. Jeffcoat, R.; Hassall, H.; Dagley, S. Biochem. J. 1969, 115, 969.
5. A partially purified GlucD from E. coli was reported to yield an 85: 15 mixture of 2 and 3 from D-glucarate and also utilize L-idarate as a substrate at a rate half of that observed for D-glucarate (Blumenthal, H. J. Methods Enzymol 1966, IX, 660).
6. 2 by a dehydratase/decarboxylase; and α-ketoglutarate semialdehyde is oxidized to α-ketoglutarate.
7. 2, and assayed by end point detection of 2 as its semicarbazone (MacGee, J.; Doudoroff, M. J. Biol. Chem. 1954, 210, 617-624).
8. 7
9. 8
10. 3 oxidation (Mehltretter, C. L. Methods Carbohydr. Chem. 1963, 2, 46-48).
11. 3 oxidation (Mehltretter, C. L. Methods Carbohydr. Chem. 1963, 2, 46-48).
12. 13C]-D-glucose (Cambridge Isotope Laboratories).
13. C6-H4 = 10.5 Hz.
14. Compounds 1 and 4 were isolated by ion exchange chromatography8 using a gradient of 0-2 M formic acid. The hexaric acids were detected as semicarbazones following dehydration with GlucD.