Process for the cell-free enzymatic production of 10-hydroxystearic acid (10-HSA) from bio-based oils for lubricant formulation

A process for cell-free enzymatic production of 10-hydroxystearic acid (10-HSA) comprising the following steps:

• 1) enzymatic hydrolysis of oil comprising at least 25% oleic acid using lipase to provide free fatty acids comprising oleic acid, and
• 2) hydration of the free fatty acids, to create a reaction mixture, using oleate-hydratase (EC 4.2.1.53), wherein the oleate-hydratase is selected from Stenotrophomonas maltophilia or derivatives thereof having at least 10% activity when compared with wild-type enzyme under the same conditions or active fragments thereof and/or Rhodococcus erythropolis or derivatives thereof having at least 10% activity when compared with wild-type enzyme under the same conditions or active fragments thereof, and
• 3) separation of 10-HSA in form of a filter cake from the reaction mixture, and
• 4) purification of 10-HSA from the separated filter cake, wherein the purification comprises extracting 10-HSA with at least one organic solvent and separating 10-HSA dissolved in the at least one organic solvent using by phase separation from a residual fraction comprising water and enzymes,
• and wherein, optionally, the lipase and/or the hydratase used in step 1) and/or step 2) is/are recycled and/or immobilised.