Hydroxycarboxylic acids may be biosynthesized from a carbonaceous feedstock and then isolated through forming and subsequently hydrolyzing an intermediate sophorolipid. After biosynthesizing a hydroxycarboxylic acid in a cell culture medium or otherwise providing a hydroxycarboxylic acid in a first aqueous medium, the hydroxycarboxylic acid and glucose may be converted into at least one sophorolipid by a suitable microorganism or an enzyme cocktail. The at least one sophorolipid may be then be separated from the cell culture medium or first aqueous medium and then hydrolyzed in a second aqueous medium to form the hydroxycarboxylic acid and glucose as free components separate from the cell culture medium or first aqueous medium. The hydroxycarboxylic acid is present as a phase separate from the second aqueous medium and the glucose remains in the second aqueous medium.