Difference between revisions of "Part:BBa K4164001"

Latest revision as of 06:59, 12 October 2022

Bile acid sulfate sulfatase（BSS）

BSS is an enzyme from Pseudomonas testosteroni and was composed of two isoforms. Each isoform was a homodimer of a subunit of which molecular weight was 53,000 or 51,000, respectively.This part is one of the protein monomers.

BSS was able to hydrolyze many kinds of bile acid 3α-sulfates and produce 3β-hydroxyl iso-compounds equivalent to each bile acid and sulfuric acid. All 3-sulfate esters of 15 kinds of bile acids routinely contained in serum, such as cholic, chenodeoxycholic, deoxycholic, lithocholic, and ursodeoxycholic acids(CDCA, DCA, LCA), and their glycine and taurine conjugates, can be hydrolyzed by it. Among these substrates, BSS has although the degrees of substrate specificities towards CDCA-S.

Fig.1 The relative activities for various substrates.MnCI₂ was added to this enzyme solution at the concentration of 1 mM.The activity for LCA-S was taken as 100%.

We used the methods described by Y. TAZUKE et al. (1992) to assay sulfatase activity. Fig.2 shows that the activity of BSS used was estimated at approximately 0.15 unit.

Fig.2 Assay of sulfatase activity. A reaction mixture containing 1.0 ml of 100 mM Tris buffer (pH 8.0), 0.20 ml of 15mM [β-NAD, 0.20ml of 1mM CDCA-S, and 1.45ml of distilled water in eppendorf tube was incubated at 30°C for 10min, and then added 0.1ml of the cell-free solution and 0.05 ml of β-HSD solution (10 units/ml).

Because of its high efficiency, BSS can be an ideal enzyme for our project.

Sequence and Features