L-aminoacylases
AssigneesMicrobial Chemistry Research Foundation, Banyu Pharmaceutical Co., Ltd.
InventorsHamao Umezawa, Tomio Takeuchi, Toshiharu Nagatsu, Masa Hamada, Shuichi Iwadare, Ikuo Matsumoto, Hajime Morishima
Life of this patent
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Abstract
L-aminoacylases S.sub.1 and S.sub.2 derived from actinomycetes and having physicochemical characteristics such that it is a L-aminoacylase which acts on a N-acyl-L-amino acid to give a L-amino acid, its substrate profile is wide, and it acts not only on a N-acyl derivative of a natural L-amino acid, but also on a N-acyl derivative of a synthetic L-amino acid, while it does not act on a N-acyl-D-amino acid, a DL-N-acetyl-.alpha.methylbenzylamine and a N-acetyl-D-glucosamine, etc.
Claims3
L-aminoacylase S.sub.1 derived from an actinomycete and having the following physicochemical characteristics:
- (1) Action and substrate profile:
- It is a L-aminoacylase which acts on a N-acyl-L-amino acid to give a L-amino acid, its substrate profile is wide, and it acts not only on a N-acyl derivative of a natural L-amino acid, but also on a N-acyl derivative of a synthetic L-amino acid, while it does not act on a N-acyl-D-amino acid, a DL-N-acetyl-.alpha.-methylbenzylamine and a N-acetyl-D-glucosamine,
- (2) Stability
- Temperature:
- it maintains its activity almost fully at 50.degree. C., and a slight activity remains even at 60.degree. C. (when left for one hour at pH 7.0),
- pH:
- it is most stable at pH 7.0-9.0, and relatively stable even at about pH 10.0, but it loses its activity at pH 4.0 or lower (when left for 24 hours at 5.degree. C.),
- (3) Temperature- and pH-dependence
- Temperature:
- its activity increases linearly up to 50.degree. C., and it loses its reactivity rapidly at a temperature of 60.degree. C. or higher,
- pH:
- its reactivity is high at pH 6.5-9.5 and the optimum pH for the reaction is about pH 7.5-8.5,
- (4) Molecular weight:
- 50,000-60,000 (gel-filtration method)
- (5) Isoelectric point:
- pI=7.00-7.70,
- (6) Disc gel electrophoresis:
- R.sub.m BPB=0.125-0.167
- (7) Influence of metal ions
- It is strongly inhibited by Fe.sup.++, Ni.sup.++, Ag.sup.+, Hg.sup.++ and Cu.sup.++, and it is activated by Co.sup.++,
- (8) Inhibitor
- It is inhibited by p-chloromercuribenzoate, it is not inhibited by monoiodoacetic acid, and it is moderately inhibited by ethylenediaminetetraacetate.
L-aminoacylase S.sub.2 derived from streptoverticellium rimofaciens MH 240-CPF7 or mutants and derivatives thereof, and having the following physiochemical characteristics:
- (1) Action and substrate profile
- It is a L-aminoacylase which acts on a N-acyl-L-amino acid to give a L-amino acid, its substrate profile is wide, and it acts not only on a N-acyl derivative of a natural L-amino acid, but also on a N-acyl derivative of a synthetic L-amino acid, while it does not act on a N-acyl-D-amino acid, a DL-N-acetyl-.alpha.-methylbenzylamine and a N-acetyl-D-glucosamine,
- (2) Stability
- Temperature:
- it does not lose its activity up to 50.degree. C., maintains 80% of the activity at 60.degree. C., and at least 20% of the activity remains even at 70.degree. C. (when left for 10 minutes at pH 7.0),
- pH:
- it is most stable at pH 8.5-10.0, and 50% of the activity remains even at about pH 3.5 and at about pH 11.0 (when left for 24 hours at 5.degree. C.),
- (3) Temperature- and pH-dependence
- Temperature:
- its relative activity increases linearly up to 60.degree. C., and it maintains at least 70% of the reactivity even at 70.degree. C.,
- pH:
- its reactivity is high at pH 7.0-10.0, the optimum pH for the reaction is about pH 8.0-9.0.
- (4) Molecular weight:
- 50,000 to 60,000 (gel-filtration method),
- (5) Isoelectric point:
- pI=6.38-7.42
- (6) Disc gel electrophoresis:
- R.sub.m BPB=0.180-0.280
- (7) Influence of metal ions
- It is strongly inhibited by Cu.sup.++, Mn.sup.++, Co.sup.++, Hg.sup.++, Fe.sup.++, Ni.sup.++ and Ag.sup.+,
- (8) Inhibitor
- It is strongly inhibited by p-chloromercuribenzoate, L-cysteine and monoidoacetic acid, but it is substantially uninhibited by ethylenediaminetetraacetate.
Dependencies are inferred from the claim text.