dimethyl cyclohexane-1,2-dicarboxylate

Conditions: temp:30.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Into a 2-L jacketed vessel set at 30° C. was placed dimethyl cyclohexane-1,2-dicarboxylate (2, 84.6 g, 0.42 mol) and 900 mL of 0.1M potassium phosphate buffer, pH 8. The mixture was continuously stirred at 30° C. and PLE (600 mg, 10200 units) was added. The mixture was stirred for 91 hours and the pH was maintained at 8 by addition of 5M NaOH solution. An aliquot from the mixture was analyzed by GC to confirm that residual starting material was less than 5%. The reaction mixture was then filtered through a Celite™ bed and the filtrate was extracted with 250 mL MTBE to remove residual starting material. The aqueous phase was then acidified to pH 4 with concentrated HCl and extracted with 3×500 mL MTBE. The combined extracts were dried over magnesium sulfate and solvent was removed under reduced pressure, to yield 69.28 g (88% yield) of (1R,2S)-2-(methoxycarbonyl)cyclohexane carboxylic acid (4) as a colorless oil with e.e.=79%. Into a 2-L jacketed vessel set at 30° C. PLE (600 mg, 10200 units) was added The mixture was stirred for 91 hours The reaction mixture was then filtered through a Celite™ bed the filtrate was extracted with 250 mL MTBE to remove residual extracted with 3×500 mL MTBE The combined extracts were dried over magnesium sulfate and solvent was removed under reduced pressure

Conditions: temp:40.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Into a 100 mL jacketed vessel was placed dimethyl cyclohexane-1,2-dicarboxylate (2, 4 g, 20 mmol) and 39 mL of 0.1M potassium phosphate buffer, pH 8. The mixture was continuously stirred at 40° C. and Novozym™ 435 (320 mg) was added. Stirring was continued at 40° C. for 43 hours and pH was maintained at 8 by addition of 2M NaOH solution. A sample from the reaction was analyzed by GC to confirm that less than 5% of the starting material remained. The reaction mixture was filtered to remove the enzyme and the filtrate was extracted with toluene (20 mL) to remove any residual starting material. The pH of the aqueous phase was readjusted to 3.5 with 2M HCl and it was extracted with 2×50 mL of MTBE. The combined extracts were dried over magnesium sulfate and concentrated under reduced pressure to yield 3.2 g (86%) of (1S,2R)-2-(methoxycarbonyl) cyclohexane carboxylic acid (3) as a colorless oil with e.e.=98%. The isolated enzyme is washed with fresh buffer for reuse with a second batch of substrate/buffer. Stirring The reaction mixture was filtered to remove the enzyme the filtrate was extracted with toluene (20 mL) to remove was extracted with 2×50 mL of MTBE The combined extracts were dried over magnesium sulfate concentrated under reduced pressure