L-phenylalanine dehydrogenase and use thereof
AssigneeSagami Chemical Research Center
InventorsYasuhisa Asano, Akiko Nakazawa, Shiro Terashima, Kiyosi Kondo, Kaori Endo, Kenji Hirai, Naganori Numao
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Abstract
New L-phenylalanine dehydrogenases produced by a microorganism belonging to the genus Sporosarcina or Bacillus, new microorganisms capable of l-phenylalanine dehydrogenase and belonging to the genus Sporosarcina or Bacillus, a process for production of L-phenylalanine dehydrogenase using the microorganisms, and processes for production of L-amino acids using the enzymes or the microorganisms.
Claims19
An enzyme L-phenylalanine dehydrogenase characterized by:
- (1) catalyzing a reaction wherein L-phenylalanine, NAD.sup.+ and H.sub.2 O react to form phenylpyruvate, NADH and ammonium ion, and a reverse reaction thereof;
- (2) having a molecular weight of about 290,000 as determined by high performance liquid chromatography and a molecular weight of about 305,000 as determined by a sedimentation equilibrium method, and exhibiting a sub-unit with a molecular weight of about 38,000 to 39,000 as determined by SDS-polyacrylamide gel electrophoresis;
- and
- (3) having a high specificity for L-phenylalanine and a very low specificity for L-tryptophan, L-tyrosine and L-methionine for oxidative deamination.
An enzyme L-phenylalanine dehydrogenase characterized by:
- (1) catalyzing a reaction wherein L-phenylalanine, NAD.sup.+ and H.sub.2 O react to form phenylpyruvate, NADH, and ammonium ion, and a reverse reaction thereof;
- and
- (2) having a molecular weight of about 290,000 as determined by high performance liquid chromatography and a molecular weight of about 305,000 as determined by a sedimentation equilibrium method, and exhibiting a sub-unit with a molecular weight of about 38,000 to 39,000 as determined by SDS-polyacrylamide gel electrophoresis;
- and
- (3) being produced by a microorganism belonging to the genus Sporosarcina.
An enzyme L-phenylalanine dehydrogenase characterized by:
- (1) catalyzing a reaction wherein L-phenylalanine, NAD.sup.+, and H.sub.2 O react to form phenylpyruvate, NADH and ammonium ion, and a reverse reaction thereof;
- (2) exhibiting a molecular weight of about 290,000 as determined by high performance liquid chromatography, a molecular weight of about 340,000 as determined by a sedimentation equilibrium method, and exhibiting a sub-unit with a molecular weight of about 38,000 to 39,000 as determined by SDS-polyacrylamide gel electrophoresis;
- and
- (3) having a high specificity for L-phenylalanine and L-tyrosine, and a very low specificity for L-tryptophan and L-methionine for oxidative deamination.
An enzyme L-phenylalanine dehydrogenase characterized by:
- (1) catalyzing a reaction wherein L-phenylalanine, NAD+ and H.sub.2 O react to form phenylpyruvate, NADH and ammonium ion;
- and
- (2) exhibiting a molecular weight of about 290,000 as determined by high performance liquid chromatography, a molecular weight of about 340,000 as determined by a sedimentation equilibrium method, and exhibiting a sub-unit with a molecular weight of about 38,000 to 39,000 as determined by SDS-polyacrylamide gel electrophoresis;
- and
- (3) being produced by a microorganism belonging to the genus Bacillus.
A process for production of L-phenylalanine dehydrogenase characterized by:
- (1) culturing a microorganism belonging to the genus Sporosarcina and being capable of producing the L-phenylalanine dehydrogenase to form the L-phenylalanine dehydrogenase;
- and
- (2) recovering the L-phenylalanine dehydrogenase.
A process for production of L-phenylalanine dehydrogenase characterized by:
- (1) culturing a microorganism belonging to the genus Bacillus and capable of producing the L-phenylalanine dehydrogenase to form the L-phenylalanine dehydrogenase;
- and
- (2) recovering the L-phenylalanine dehydrogenase.
A biologically pure culture of microorganism capable of producing L-phenylalanine dehydrogenase, which is Sporosarcina ureae SCRC-R04, FERM P-8178, FERM BP-1012.
A biologically pure culture of microorganism capable of producing L-phenylalanine dehydrogenase, selected from the group consisting of Bacillus sp. SCRC-R79a FERM P-8179, FERM BP-1013;
- and Bacillus sp. SCRC-114D, FERM BP-1011.
A process for production of L-amino acid having the general formula (I):
- ##STR3## wherein R.sub.1 represents hydrogen, or methyl;
- and R.sub.2 represents a linear or branched optionally substituted alkyl having 1 to 4 carbon atoms, or an optionally substituted aromatic group,
- characterized by reacting a .alpha.-ketocarboxylic acid having the general formula (II):
- ##STR4## wherein R.sub.1 and R.sub.2 are the same as defined above, or a salt thereof, ammonium ion, and NADH in the presence of L-phenylalanine dehydrogenase derived from microorganism belonging to the species Sporosarcina ureae or the genus Bacillus to form L-amino acid, and recovering the L-amino acid.
A process for production of L-amino acid having the general formula (I):
- ##STR5## wherein R.sub.1 represents hydrogen, or methyl;
- and R.sub.2 represents a linear or branched optionally substituted alkyl having 1 to 4 carbon atoms, or an optionally substituted aromatic group, characterized by reacting a .alpha.-ketocarboxylic acid having the general formula (II):
- ##STR6## wherein R.sub.1 and R.sub.2 are the same as defined above, or a salt thereof, and an ammonium ion in the presence of cultured broth, living cells or cells treated in an extent wherein enzyme systems necessary for the conversion of .alpha.-ketocarboxylic acid to a corresponding L-amino acid remain, derived from a microorganism belonging to the species Sporosarcina ureae and capable of producing L-phenylalanine dehydrogenase, and in the presence of an energy source to form L-amino acid, wherein the L-phenylalanine dehydrogenase characterized by:
- (1) catalyzing a reaction wherein L-phenylalanine, NAD.sup.+ and H.sub.2 O react to form phenylpyruvate, NADH and ammonium ion, and reverse reaction thereof;
- and
- (2) having a molecular weight of about 290,000 as determined by high performance liquid chromatography and a molecular weight of about 305,000 as determined by a sedimentation equilibrium method, and exhibiting a sub-unit with a molecular weight of about 38,000 to 39,000 as determined by SDS-polyacrylamide gel electrophoresis.
Dependencies are inferred from the claim text.