(1S,2R)-2-methoxycarbonylcyclohexane-1-carboxylic acid

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

Procedure: Into a 25 mL jacketed vessel cooled to below 0° C. was placed a solution of (1S,2R)-2-(methoxycarbonyl) cyclohexane carboxylic acid (3, 880 mg, 4.72 mmol) and triethylamine 659 μL, 4.72 mmol) in THF (6.6 mL). A solution of ethyl chloroformate (512 μL, 4.72 mmol) in 1.2 mL of THF was added slowly over a few minutes and the resulting mixture was stirred for 30 minutes. The precipitated triethylamine hydrochloride salt was removed by filtration and the filtrate was added drop-wise to a suspension of sodium borohydride in 4.6 mL of water at 12° C. After the addition was complete, the reaction mixture was stirred at 20° C. for a further 3.5 hours. The reaction mixture was then cooled to below 10° C., acidified to pH 4 with 2M HCl solution, and extracted with 2×15 mL of dichloromethane. The combined organic extracts were dried over magnesium sulfate and solvent was removed under reduced pressure, to yield 450 mg of a colorless oil. The material was purified by short path distillation to yield 170 mg of colorless oil with e.e.=98% and [α]D20=−39.3° (c 1, methanol). Into a 25 mL jacketed vessel cooled to below 0° C. The precipitated triethylamine hydrochloride salt was removed by filtration the filtrate was added drop-wise to a suspension of sodium borohydride in 4.6 mL of water at 12° C After the addition the reaction mixture was stirred at 20° C. for a further 3.5 hours The reaction mixture was then cooled to below 10° C. extracted with 2×15 mL of dichloromethane The combined organic 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