Expression and Stereochemical and Isotope Effect Studies of Active 4-Oxalocrotonate Decarboxylase

• Published in: Biochemistry (Easton) Vol. 39; no. 4; pp. 718 - 726
• Main Authors: Stanley, Thanuja M, Johnson, William H, Burks, Elizabeth A, Whitman, Christian P, Hwang, Chi-Ching, Cook, Paul F
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.02.2000
• Subjects: 4-Oxalocrotonate decarboxylase; Carbon Isotopes; Carboxy-Lyases - biosynthesis; Carboxy-Lyases - chemistry; Carboxy-Lyases - genetics; Deuterium; Enzyme Activation - genetics; Escherichia coli - genetics; Genetic Vectors - chemical synthesis; Glutamic Acid - chemistry; Glutamic Acid - genetics; Glutamine - chemistry; Glutamine - genetics; Hydro-Lyases - genetics; Kinetics; Mutagenesis, Site-Directed; Nuclear Magnetic Resonance, Biomolecular; Protons; Pseudomonas putida; Pseudomonas putida - enzymology; Pseudomonas putida - genetics; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemical synthesis; Recombinant Proteins - isolation & purification; Stereoisomerism

4-Oxalocrotonate decarboxylase (4-OD) and vinylpyruvate hydratase (VPH) from Pseudomonas putida mt-2 form a complex that converts 2-oxo-3-hexenedioate to 2-oxo-4-hydroxypentanoate in the catechol meta fission pathway. To facilitate mechanistic and structural studies of the complex, the two enzymes h...