Purification and Characterization of Amino Acid Racemase with Very Broad Substrate Specificity from Aeromonas caviae

Kenji Inagaki, Katsuyuki Tanizawa, Hidehiko Tanaka, Kenji Soda

Research output: Contribution to journalArticle

Abstract

A bacterium which grows in a medium containing D-serine as a sole nitrogen source has been isolated from soil and identified as Aeromonas caviae. The bacterium had a high enzyme activity catalyzing racemization of various amino acids. The enzyme, purified to homogeneity by polyacrylamide gel electrophoresis and ultracentrifugation, has a molecular weight of about 76,000, and is composed of two subunits identical in molecular weight (40,000). The results of enantiomeric analysis of the reaction product and kinetic examination of the reaction indicate that the enzyme catalyzes the complete racemization of substrates. The enzyme has absorption maxima at 280 and 420 nm, and contains 2 mol of pyridoxal 5’-phosphate per mol of enzyme. The holoenzyme is resolved to the apoenzyme by treatment with hydroxylamine, and reconstituted by the addition of pyridoxal 5’-phosphate. The very broad substrate specificity of the A. caviae amino acid racemase is comparable to that of the Pseudomonas striata enzyme. However, they share no common antigenic determinants.

Original languageEnglish
Pages (from-to)173-180
Number of pages8
JournalAgricultural and Biological Chemistry
Volume51
Issue number1
DOIs
Publication statusPublished - 1987
Externally publishedYes

Fingerprint

Amino Acid Isomerases
Aeromonas caviae
Aeromonas punctata
substrate specificity
Substrate Specificity
Purification
pyridoxal phosphate
amino acids
Substrates
Enzymes
enzymes
Pyridoxal Phosphate
molecular weight
Bacteria
apoproteins
Molecular weight
ultracentrifugation
Pseudomonas putida
bacteria
enzymatic reactions

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Purification and Characterization of Amino Acid Racemase with Very Broad Substrate Specificity from Aeromonas caviae. / Inagaki, Kenji; Tanizawa, Katsuyuki; Tanaka, Hidehiko; Soda, Kenji.

In: Agricultural and Biological Chemistry, Vol. 51, No. 1, 1987, p. 173-180.

Research output: Contribution to journalArticle

@article{1717877f17024a5ab36025eea873f013,
title = "Purification and Characterization of Amino Acid Racemase with Very Broad Substrate Specificity from Aeromonas caviae",
abstract = "A bacterium which grows in a medium containing D-serine as a sole nitrogen source has been isolated from soil and identified as Aeromonas caviae. The bacterium had a high enzyme activity catalyzing racemization of various amino acids. The enzyme, purified to homogeneity by polyacrylamide gel electrophoresis and ultracentrifugation, has a molecular weight of about 76,000, and is composed of two subunits identical in molecular weight (40,000). The results of enantiomeric analysis of the reaction product and kinetic examination of the reaction indicate that the enzyme catalyzes the complete racemization of substrates. The enzyme has absorption maxima at 280 and 420 nm, and contains 2 mol of pyridoxal 5’-phosphate per mol of enzyme. The holoenzyme is resolved to the apoenzyme by treatment with hydroxylamine, and reconstituted by the addition of pyridoxal 5’-phosphate. The very broad substrate specificity of the A. caviae amino acid racemase is comparable to that of the Pseudomonas striata enzyme. However, they share no common antigenic determinants.",
author = "Kenji Inagaki and Katsuyuki Tanizawa and Hidehiko Tanaka and Kenji Soda",
year = "1987",
doi = "10.1271/bbb1961.51.173",
language = "English",
volume = "51",
pages = "173--180",
journal = "Bioscience, Biotechnology and Biochemistry",
issn = "0916-8451",
publisher = "Japan Society for Bioscience Biotechnology and Agrochemistry",
number = "1",

}

TY - JOUR

T1 - Purification and Characterization of Amino Acid Racemase with Very Broad Substrate Specificity from Aeromonas caviae

AU - Inagaki, Kenji

AU - Tanizawa, Katsuyuki

AU - Tanaka, Hidehiko

AU - Soda, Kenji

PY - 1987

Y1 - 1987

N2 - A bacterium which grows in a medium containing D-serine as a sole nitrogen source has been isolated from soil and identified as Aeromonas caviae. The bacterium had a high enzyme activity catalyzing racemization of various amino acids. The enzyme, purified to homogeneity by polyacrylamide gel electrophoresis and ultracentrifugation, has a molecular weight of about 76,000, and is composed of two subunits identical in molecular weight (40,000). The results of enantiomeric analysis of the reaction product and kinetic examination of the reaction indicate that the enzyme catalyzes the complete racemization of substrates. The enzyme has absorption maxima at 280 and 420 nm, and contains 2 mol of pyridoxal 5’-phosphate per mol of enzyme. The holoenzyme is resolved to the apoenzyme by treatment with hydroxylamine, and reconstituted by the addition of pyridoxal 5’-phosphate. The very broad substrate specificity of the A. caviae amino acid racemase is comparable to that of the Pseudomonas striata enzyme. However, they share no common antigenic determinants.

AB - A bacterium which grows in a medium containing D-serine as a sole nitrogen source has been isolated from soil and identified as Aeromonas caviae. The bacterium had a high enzyme activity catalyzing racemization of various amino acids. The enzyme, purified to homogeneity by polyacrylamide gel electrophoresis and ultracentrifugation, has a molecular weight of about 76,000, and is composed of two subunits identical in molecular weight (40,000). The results of enantiomeric analysis of the reaction product and kinetic examination of the reaction indicate that the enzyme catalyzes the complete racemization of substrates. The enzyme has absorption maxima at 280 and 420 nm, and contains 2 mol of pyridoxal 5’-phosphate per mol of enzyme. The holoenzyme is resolved to the apoenzyme by treatment with hydroxylamine, and reconstituted by the addition of pyridoxal 5’-phosphate. The very broad substrate specificity of the A. caviae amino acid racemase is comparable to that of the Pseudomonas striata enzyme. However, they share no common antigenic determinants.

UR - http://www.scopus.com/inward/record.url?scp=85008122310&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85008122310&partnerID=8YFLogxK

U2 - 10.1271/bbb1961.51.173

DO - 10.1271/bbb1961.51.173

M3 - Article

AN - SCOPUS:85008122310

VL - 51

SP - 173

EP - 180

JO - Bioscience, Biotechnology and Biochemistry

JF - Bioscience, Biotechnology and Biochemistry

SN - 0916-8451

IS - 1

ER -