Two extracellular lipases from Bacillus subtilis, B. subtilis lipase A and lipase B, have been expressed in the heterologous host Escherichia coli, biochemically characterized and used for the kinetic resolution of (rac)-trans-1,2-diacetoxycyclohexane. Both enzymes were selectively acting on the (R,R)-enantiomer of the racemic substrate, highly specifically hydrolyzing only one of the two ester groups present, thus allowing the preparation of enantiopure (R,R)- and (S,S)-cyclohexane-trans-1,2-diol. The reaction conditions for the use of purified enzyme and crude cell lyophilizate were optimized and reactions in batch and repetitive batch modes were carried out on a preparative scale to yield enantiopure product (>99% enantiomeric excess).

The efficacy of the β-and δ-amino acids (5S,6S)-6-amino-5-hydroxycyclohexa-1,3-dienecarboxylic acid (2,3-trans-CHA) and (3R,4R)-4-amino-3-hydroxycyclohexa-1,5-dienecarboxylic acid (3,4-trans-CHA) as catalysts in Knoevenagel condensation and aldol addition reactions is studied. Synthesis of the zinc(II) complexes of 2,3-and 3,4-trans-CHA provided precipitated material of sufficient purity for use. The catalytic Knoevenagel reactions were carried out with the β-amino acid 2,3-trans-CHA and the δ-amino acid 3,4-trans-CHA, resulting in a product yield of up to 61%. The asymmetric aldol addition reactions were carried out with catalytic amounts of the zinc(II) complexes of 2,3-and 3,4-trans-CHA. In this case, it was observed that the ee of the product (up to 90%) depends on the conversion and/or the reaction time.