Inventors: Mosbach K
, Staahl M
, Maansson M-O
Synthesis of D-aminoacidester in an organic solvent - using chymotrypsin modified by bio-molecular imprinting method
Publication date: 1991
Crystalline alpha-chymotrypsin from bovine pancreas of type II (EC 188.8.131.52) is used as a freeze-dried powder, having a specific activity of 46 U/mg with N-benzoyl-L-tyrosine ethyl ester as substrate, pH 7.8 and 25 deg.C. - 0 mM of sodium phosphate buffered with pH 7.8 contg. 20 mM of either N-acetyl-D-tryptophan, corresp. L-isomers or without binder and enzyme soln. cooled to 0 deg.C 1-propanol(4 ml) is added at a temp. of -20 deg.C. - Pptn. is ensured by centrifuging after 30 minutes on ice. After repeated washing with clean 1-propanol, the appts. is dried under vacuum, for 4 hours. The chymotrypsin pptn. is then suspended in cyclohexane (4 mg/ml) and treated to reduce particle size. The adhesive is commenced by adding the substrate dissolved in 99.5 per cent ethanol.; The final soln. is found to contain 80 per cent cyclohexane, 20 mM substrate (N-acetyl-D-tryptophan or L-isomers), 3.4 M ethanol and 8 mg enzyme in a total volume of 2.5 ml. The content in the system is estimated at a max. of 0.1 per cent.
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