Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing alpha or beta amino acid or substituted amino acid...
Patent
1994-06-06
1999-05-11
Saucier, Sandra E.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing alpha or beta amino acid or substituted amino acid...
435280, 435874, 435911, C12P13/04
Patent
active
059027363
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to a process for the production of a D-.alpha.-amino acid, in which a D-N-carbamyl-.alpha.-amino acid corresponding to the general formula: ##STR3## wherein R represents phenyl, hydroxy-substituted phenyl, substituted or unsubstituted alkyl preferably having 1 to 5 carbon atoms, or thienyl, is converted by an enzyme having an ability to eliminate the carbamyl group (hereinafter referred to as "decarbamylase") to a D-.alpha.-amino acid corresponding to the formula: ##STR4## wherein R is the same as defined above.
These optically active D-.alpha.-amino acids are important as pharmaceutical intermediates. In particular, D-phenylglycine and D-(4-hydroxyphenyl)glycine (hereinafter referred to as "D-HPG") are useful for the production of semi-synthetic penicillin and semi-synthetic cephalosporin.
PRIOR ART
Production of D-.alpha.-amino acids by eliminating carbamyl groups from corresponding D-N-carbamyl-.alpha.-amino acids has been known. The elimination has been done chemically (Japanese Patent Publication No. 4707/1983) or by enzymatic reactions of microorganisms (Japanese Patent Publication Nos. 18793/1982, 20520/1988 and 48758/1989).
Most of these microbial enzymes have optimum pH values around a neutral range. However, in the previous step, wherein the N-carbamyl-D-.alpha.-amino acids are prepared by selectively hydrolyzing the D-form of corresponding DL-5-substituted hydantoins, hydantoin hydrolase having optimum pH of from 8 to 9 is used. Therefore, the subsequent conversion of the N-carbamyl-D-.alpha.-amino acids to D-.alpha.-amino acids by decarbamylase has to be carried out in a different reaction medium from that in the previous step.
SUMMARY OF THE INVENTION
The present inventors made a search for microorganisms which could enzymatically eliminate carbamyl groups ("decarbamylation") from D-N-carbamyl-.alpha.-amino acids to form D-.alpha.-amino acids, mainly among the genera which are unknown to have decarbamylase. It has now been found that the bacteria of the genera Comamonas, Blastobacter, Alcaligenes, Sporosarcina, Rhizobium, Bradyrhizobium and Arthrobacter have the decarbamylase activity.
Accordingly, the present invention relates to a process for the production of a D-.alpha.-amino acid, in which an N-carbamyl-D-.alpha.-amino acid corresponding to the general formula (I): ##STR5## wherein R represents phenyl, hydroxy-substituted phenyl, substituted or unsubstituted alkyl preferably having 1 to 5 carbon atoms, or thienyl, is converted by a microbial enzyme in an aqueous medium to a D-.alpha.-amino acid corresponding to the general formula (II): ##STR6## wherein R is the same as defined above, characterized in that the enzyme is decarbamylase produced by a microorganism of the genus Comamonas, Blastobacter, Alcaligenes, Sporosarcina, Rhizobium, Bradyrhizobium or Arthrobacter.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a graph showing the results of decarbamylation at 50.degree. C. and 30.degree. C. by newly screened strains.
FIG. 2 is a graph showing the effects of various additives to the culture medium of Comamonas sp. E 222 C on its decarbamylase productivity.
FIG. 3 is a result of SDS-polyacrylamide gel electrophoresis of purified Comamonas sp. E 222 C decarbamylase.
FIG. 4 is a result of gel filtration of the purified Comamonas sp. E 222 C decarbamylase thorough a Sephadex G-150 column.
FIG. 5 is a graph showing the effect of pH on the activity of the purified Comamonas sp. E 222 C decarbamylase.
FIG. 6 is a graph showing the effect of temperature on the activity of the purified Comamonas sp. E 222 C decarbamylase.
FIG. 7 is a graph showing the effects of various additives to the culture medium of Blastobacter sp. A 17 p-4 on its decarbamylase productivity.
FIG. 8 is a graph showing the effects of various metal ions added to the culture medium of Blastobacter sp. A 17 p-4 on its decarbamylase productivity.
FIG. 9 is a result of SDS-polyacrylamide gel electrophoresis of purified Blastobacter sp. A 17 p-4 decarbamylase.
FIG. 10 is
REFERENCES:
patent: 4211840 (1980-07-01), Nakamori et al.
patent: 4242452 (1980-12-01), Yamada et al.
patent: 4312948 (1982-01-01), Olivieri et al.
Nichols HF et al, Plant Growth Regal. 12(3) 257-44 (1993).
Ogawa J et al, J. Biotechnol 38:11-19 (1994).
Ogawa J et al, Eur. J. Biochem 212:685-9 (1993).
ATCC Catalogue of Bacteria and Bacteriophages p. 1992.
Yokozeki et al, Agric. Biol. Chem 51:721-8 (1987).
Ikenaka Yasuhiro
Nanba Hirokazu
Shimizu Sakayu
Takahashi Satomi
Takano Masayuki
Kanegafuchi Chemical Industry Co. Ltd.
Saucier Sandra E.
LandOfFree
Process for the production of D-.alpha.-amino acids by hydrolysi does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Process for the production of D-.alpha.-amino acids by hydrolysi, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Process for the production of D-.alpha.-amino acids by hydrolysi will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-245269