Purification and characterization of l-alanine: 4,5-dioxovalerate (glyoxylate) aminotransferase from radish (raphanus sativus l.) seedlings

Yuzo Shioi, Michio Doi, Tsutomu Sasa

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

L-Alanine:4,5-dioxovalerate (DOVA) aminotransferase was purified 131-fold and characterized from greening seedlings of radish (Raphanus sativus L.). The enzyme was shown to be identical with alanine:glyoxylate aminotransferase. The rate of activity of DOVA aminotransferase was 15 times less than that of glyoxylate aminotransferase. Its molecular weight was estimated to be approximately 123,000 with two identical subunits, and exhibited a single, broad pH optimum at 8.0. DOVA aminotransferase activity was competitively inhibited by glyoxylate. A kinetic study of the enzyme at different alanine concentrations suggested a ping pong reaction mechanism. The Km values for DOVA and L-alanine were 0.71 and 1.7 mM, respectively.The activity ratio of transamination under various conditions, the cellular localization of the enzyme and the lack of correlation between the activity of this enzyme and chlorophyll synthesis, indicate that DOVA aminotransferase in radish is not involved in 5-aminolevulinate synthesis.

Original languageEnglish
Pages (from-to)1487-1493
Number of pages7
JournalPlant and Cell Physiology
Volume25
Issue number8
DOIs
Publication statusPublished - Jan 1 1984

Fingerprint

Raphanus
Raphanus sativus
transaminases
radishes
Transaminases
Seedlings
Alanine
alanine
seedlings
Enzymes
Aminolevulinic Acid
Chlorophyll
enzymes
transamination
synthesis
reaction mechanisms
Molecular Weight
glyoxylate aminotransferase
enzyme activity
molecular weight

All Science Journal Classification (ASJC) codes

  • Physiology
  • Plant Science
  • Cell Biology

Cite this

Purification and characterization of l-alanine : 4,5-dioxovalerate (glyoxylate) aminotransferase from radish (raphanus sativus l.) seedlings. / Shioi, Yuzo; Doi, Michio; Sasa, Tsutomu.

In: Plant and Cell Physiology, Vol. 25, No. 8, 01.01.1984, p. 1487-1493.

Research output: Contribution to journalArticle

@article{bb0f83f93b044da69d803658c945323c,
title = "Purification and characterization of l-alanine: 4,5-dioxovalerate (glyoxylate) aminotransferase from radish (raphanus sativus l.) seedlings",
abstract = "L-Alanine:4,5-dioxovalerate (DOVA) aminotransferase was purified 131-fold and characterized from greening seedlings of radish (Raphanus sativus L.). The enzyme was shown to be identical with alanine:glyoxylate aminotransferase. The rate of activity of DOVA aminotransferase was 15 times less than that of glyoxylate aminotransferase. Its molecular weight was estimated to be approximately 123,000 with two identical subunits, and exhibited a single, broad pH optimum at 8.0. DOVA aminotransferase activity was competitively inhibited by glyoxylate. A kinetic study of the enzyme at different alanine concentrations suggested a ping pong reaction mechanism. The Km values for DOVA and L-alanine were 0.71 and 1.7 mM, respectively.The activity ratio of transamination under various conditions, the cellular localization of the enzyme and the lack of correlation between the activity of this enzyme and chlorophyll synthesis, indicate that DOVA aminotransferase in radish is not involved in 5-aminolevulinate synthesis.",
author = "Yuzo Shioi and Michio Doi and Tsutomu Sasa",
year = "1984",
month = "1",
day = "1",
doi = "10.1093/oxfordjournals.pcp.a076861",
language = "English",
volume = "25",
pages = "1487--1493",
journal = "Plant and Cell Physiology",
issn = "0032-0781",
publisher = "Oxford University Press",
number = "8",

}

TY - JOUR

T1 - Purification and characterization of l-alanine

T2 - 4,5-dioxovalerate (glyoxylate) aminotransferase from radish (raphanus sativus l.) seedlings

AU - Shioi, Yuzo

AU - Doi, Michio

AU - Sasa, Tsutomu

PY - 1984/1/1

Y1 - 1984/1/1

N2 - L-Alanine:4,5-dioxovalerate (DOVA) aminotransferase was purified 131-fold and characterized from greening seedlings of radish (Raphanus sativus L.). The enzyme was shown to be identical with alanine:glyoxylate aminotransferase. The rate of activity of DOVA aminotransferase was 15 times less than that of glyoxylate aminotransferase. Its molecular weight was estimated to be approximately 123,000 with two identical subunits, and exhibited a single, broad pH optimum at 8.0. DOVA aminotransferase activity was competitively inhibited by glyoxylate. A kinetic study of the enzyme at different alanine concentrations suggested a ping pong reaction mechanism. The Km values for DOVA and L-alanine were 0.71 and 1.7 mM, respectively.The activity ratio of transamination under various conditions, the cellular localization of the enzyme and the lack of correlation between the activity of this enzyme and chlorophyll synthesis, indicate that DOVA aminotransferase in radish is not involved in 5-aminolevulinate synthesis.

AB - L-Alanine:4,5-dioxovalerate (DOVA) aminotransferase was purified 131-fold and characterized from greening seedlings of radish (Raphanus sativus L.). The enzyme was shown to be identical with alanine:glyoxylate aminotransferase. The rate of activity of DOVA aminotransferase was 15 times less than that of glyoxylate aminotransferase. Its molecular weight was estimated to be approximately 123,000 with two identical subunits, and exhibited a single, broad pH optimum at 8.0. DOVA aminotransferase activity was competitively inhibited by glyoxylate. A kinetic study of the enzyme at different alanine concentrations suggested a ping pong reaction mechanism. The Km values for DOVA and L-alanine were 0.71 and 1.7 mM, respectively.The activity ratio of transamination under various conditions, the cellular localization of the enzyme and the lack of correlation between the activity of this enzyme and chlorophyll synthesis, indicate that DOVA aminotransferase in radish is not involved in 5-aminolevulinate synthesis.

UR - http://www.scopus.com/inward/record.url?scp=77957178829&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77957178829&partnerID=8YFLogxK

U2 - 10.1093/oxfordjournals.pcp.a076861

DO - 10.1093/oxfordjournals.pcp.a076861

M3 - Article

AN - SCOPUS:77957178829

VL - 25

SP - 1487

EP - 1493

JO - Plant and Cell Physiology

JF - Plant and Cell Physiology

SN - 0032-0781

IS - 8

ER -