Purification and Properties of a Novel Sulfatase from Pseudomonas testosteroni That Hydrolyzed 3β-Hydroxy-5-cholenoic Acid 3-Sulfate

Abstract

  A novel sulfatase hydrolyzing the sulfate ester bond in 3β-hydroxy-5-cholenoic acid 3-sulfate (Δ⁵-3β-sulfate) was purified from Pseudomonas testosteroni ATCC 11996 as the second bile acid sulfatase. The molecular weight was 95,000 and the molecule was composed of a homodimer of a subunit of which the molecular weight was 46,000. This sulfatase hydrolyzed Δ⁵-3β-sulfate to 3α-hydroxy-5-cholenoic acid and sulfuric acid with inversion of β- to α-configuration of the hydroxyl group at the C-3 position of the substrate. The optimum pH and the stable pH of the enzyme were 8.5 and 6.5-9.7, respectively. 3β-Sulfate ester bonds of steroids such as isolithocholic acid, pregnenolone, and epiandrosterone, in which the side chain of the steroid ring was shorter than cholesterol, were also hydrolyzed to 3α-hydroxyl compounds corresponding to each steroid compound and sulfuric acid. We tentatively named this novel enzyme bile acid 3β-sulfate sulfohydrolyase (β-BSS).