Related Experiment Videos
Arylsulphatase from Alteromonas carrageenovora
T Barbeyron1, P Potin, C Richard
1Centre d'Etudes d'Océanologie et de Biologie marine, CNRS UPR, Roscoff, France.
Microbiology (Reading, England)
|November 1, 1995
Summary
Marine bacteria Alteromonas carrageenovora possess arylsulphatase activity, with its production not repressed by sulphate. The cloned arylsulphatase gene (atsA) and enzyme characteristics offer insights into glycosulphohydrolases.
Area of Science:
- Microbiology
- Enzymology
- Molecular Biology
Background:
- Marine bacterium Alteromonas carrageenovora exhibits arylsulphatase activity.
- Unlike most microbial arylsulfatases, its production is not repressed by sulfate.
Purpose of the Study:
- To clone and sequence the arylsulphatase gene (atsA) from A. carrageenovora.
- To characterize the enzyme's biochemical and structural properties.
- To explore functional analogies with other microbial arylsulfatases.
Main Methods:
- Gene cloning and sequencing of the atsA gene.
- Partial purification of arylsulphatase from A. carrageenovora and recombinant E. coli.
- Biochemical characterization including pI, Michaelis constant (Km), and molecular mass determination (SDS-PAGE).
- Bioinformatic analysis using hydrophobic cluster analysis for secondary structure comparison.
Main Results:
- The atsA gene consists of an ORF of 984 bp, encoding a 328-amino acid protein (35,797 Da).
- Both native and recombinant arylsulphatases share a pI of 5.5, Km of 68 µM for methylumbelliferyl sulphate, and a molecular mass of ~35 kDa.
- Hydrophobic cluster analysis suggests functional similarity to arylsulfatases from Mycobacterium leprae and a protein from Porphyromonas gingivalis.
Conclusions:
- The arylsulphatase from A. carrageenovora is a novel enzyme whose production is sulfate-independent.
- The enzyme and its gene have been characterized, providing a basis for further structural and functional studies.
- These findings suggest a conserved role for such enzymes as glycosulphohydrolases in desulfating polysaccharides across different microbial species.