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Sulphoacetaldehyde sulpho-lyase (EC 4.4.1.12) from Desulfonispora thiosulfatigenes: purification, properties and primary sequence.
- Source :
-
The Biochemical journal [Biochem J] 2001 Jul 15; Vol. 357 (Pt 2), pp. 581-6. - Publication Year :
- 2001
-
Abstract
- The strictly anaerobic bacterium Desulfonispora thiosulfatigenes ferments taurine via sulphoacetaldehyde, which is hydrolysed to acetate and sulphite by sulphoacetaldehyde sulpho-lyase (EC 4.4.1.12). The lyase was expressed at high levels and a two-step, 4.5-fold purification yielded an apparently homogeneous soluble protein, which was presumably a homodimer in its native form; the molecular mass of the subunit was about 61 kDa (by SDS/PAGE). The mass was determined to be 63.8 kDa by matrix-assisted laser-desorption ionization-time-of-flight (MALDI-TOF) MS. The purified enzyme converted 1 mol of sulphoacetaldehyde to 1 mol each of sulphite and acetate, but no requirement for thiamine pyrophosphate (TPP) was detected. The N-terminal and two internal amino acid sequences were determined, which allowed us to generate PCR primers. The gene was amplified and sequenced. The DNA sequence had no significant homologue in the databases searched, whereas the derived amino acid sequence indicated an oxo-acid lyase, revealed a TPP-binding site and gave a derived molecular mass of 63.8 kDa.
- Subjects :
- Amino Acid Sequence
Bacteria, Anaerobic genetics
Bacteria, Anaerobic growth & development
Chromatography, Ion Exchange
Electrophoresis, Polyacrylamide Gel
Gram-Positive Bacteria genetics
Gram-Positive Bacteria growth & development
Kinetics
Lyases genetics
Molecular Sequence Data
Molecular Weight
Bacteria, Anaerobic enzymology
Gram-Positive Bacteria enzymology
Lyases chemistry
Lyases metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0264-6021
- Volume :
- 357
- Issue :
- Pt 2
- Database :
- MEDLINE
- Journal :
- The Biochemical journal
- Publication Type :
- Academic Journal
- Accession number :
- 11439112
- Full Text :
- https://doi.org/10.1042/0264-6021:3570581