| Autor: |
Saavedra CP; Laboratorio de Microbiología Molecular, Facultad de Química y Biología, Universidad de Santiago de Chile, Casilla 40, Correo 33, Santiago, Chile., Encinas MV, Araya MA, Pérez JM, Tantaleán JC, Fuentes DE, Calderón IL, Pichuantes SE, Vásquez CC |
| Jazyk: |
angličtina |
| Zdroj: |
Biochimie [Biochimie] 2004 Jul; Vol. 86 (7), pp. 481-5. |
| DOI: |
10.1016/j.biochi.2004.06.003 |
| Abstrakt: |
The cysK gene encoding a cysteine synthase of Geobacillus stearothermophilus V was overexpressed in E. coli and the recombinant protein was purified and characterized. The enzyme is a thermostable homodimer (32 kDa/monomer) belonging to the beta family of pyridoxal phosphate (PLP)-dependent enzymes. UV-visible spectra showed absorption bands at 279 and 410 nm. The band at 279 nm is due to tyrosine residues as the enzyme lacks tryptophan. The 410 nm band represents absorption of the coenzyme bound as a Schiff base to a lysine residue of the protein. Fluorescence characteristics of CysK's Schiff base were influenced by temperature changes suggesting different local structures at the cofactor binding site. The emission of the Schiff base allowed the determination of binding constants for products at both 20 degrees C and 50 degrees C. At 50 degrees C and in the absence of sulphide the enzyme catalyzes the decomposition of O-acetyl-l-serine to pyruvate and ammonia. At 20 degrees C, however, a stable alpha-aminoacrylate intermediate is formed. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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