D-aminoacylases, method for producing the same, and method for producing D-amino acids using the same
AssigneeDaicel Chemical Industries, Ltd., a Japan corporation
InventorsKazuya Mitsuhashi, Hiroaki Yamamoto, Akinobu Matsuyama, Shinji Tokuyama
Life of this patent
Estimating the term…
Abstract
D-aminoacylase derived from fungi is provided. The fungi capable of producing D-aminoacylase include those belonging to the genus Hypomyces, Fusarium, Auricularia, Pythium, and Menisporopsis. The fungal D-aminoacylase is useful for efficiently producing D-amino acids from N-acetyl-D-amino acids.
Claims25
A D-aminoacylase derived from a fungus.
A D-aminoacylase, which has the physico-chemical properties (a) through (f) below:
- (a) function:
- the enzyme acts on N-acetyl-D-amino acids to produce corresponding D-amino acids;
- (b) molecular weight:
- the molecular weight of the enzyme is estimated to be about 64,000 daltons by SDS-polyacrylamide gel electrophoresis, and about 56,000 daltons by gel filtration chromatography on Superdex 200 Hi-Load 6/16 (Amersham Pharmacia Biotech);
- (c) substrate specificity:
- the enzyme acts on N-acetyl-D-tryptophan, N-acetyl-D-phenylalanine, N-acetyl-D-valine, N-acetyl-D-leucine, and N-acetyl-D-methionine, but not on N-acetyl-L-tryptophan, N-acetyl-L-phenylalanine, N-acetyl-L-valine, N-acetyl-L-leucine, or N-acetyl-L-methionine;
- (d) thermostability:
- when heated at pH 9.5 for 30 min, the enzyme is stable at 45° C., but inactivated at higher than 60° C.;
- (e) optimal temperature for activity:
- for the reaction at pH 7.5, the enzyme activity is optimal at about 45° C.;
- and
- (f) stabilizer:
- the enzyme activity is stabilized by reducing agents, and further activated by ICH2CONH2.
A method for producing D-amino acids, wherein said method comprises reacting a fungus capable of producing D-aminoacylase or D-aminoacylase derived from a fungus with N-acyl-DL-amino acid or its salt represented by the formula (I):
- 3
- wherein R1 and R2 may be identical or different and each represents a hydrogen atom or a substituted or unsubstituted hydrocarbon group, provided that R2 does not represent a hydrogen atom;
- and X is H, NH4, or a metal ion.
A method for producing D-aminoacylase, wherein said method comprises culturing a fungus.
Dependencies are inferred from the claim text.