PURIFICATION AND PROPERTIES OF A NOVEL SULFATASE FROM PSEUDOMONAS-TESTOSTERONI THAT HYDROLYZED 3-BETA-HYDROXY-5-CHOLENOIC ACID 3-SULFATE

A novel sulfatase hydrolyzing the sulfate ester bond in 3 beta-hydroxy -5-cholenoic acid 3-sulfate (Delta(5)-3 beta-sulfate) was purified fro m Pseudomonas testosteroni ATCC 11996 as the second bile acid sulfatas e. 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 Delta(5)-3 beta-sulfate to 3 alpha-hydroxy-5-chol enoic acid and sulfuric acid with inversion of beta- to alpha-configur ation 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, respe ctively. 3 beta-Sulfate ester bonds of steroids such as isolithocholic acid, pregnenolone, and epiandrosterone, in which the side chain of t he steroid ring was shorter than cholesterol, were also hydrolyzed to 3 alpha-hydroxyl compounds corresponding to each steroid compound and sulfuric acid. We tentatively named this novel enzyme bile acid 3 beta -sulfate sulfohydrolyase (beta-BSS).