Purification and characterization of benzoyl-l-arginine p-nitroanilide hydrolase from etiolated leaves of Zea mays

Michio Doi, Yuzo Shioi, Tsutomu Sasa

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Benzoyl-l-arginine p-nitroanilide hydrolase in the etiolated leaves of Zea mays L. has been purified 1,266-fold by a combination of gel filtration, ion exchange, and hydrophobic chromatography with a recovery of 13%. The specific activity of the purified enzyme is 5.7 units/mg protein. The enzyme is an acidic protein with a pI value of 4.6 and optimum pH of 8.2. The molecular weight of the enzyme was estimated to be 59,000. Substrate inhibition was observed at a concentration higher than 30 μM BAPA and the apparent Km for BAPA was 29 μM at pH 8.0. The enzyme activity was inhibited by sulfhydryl reagents, leupeptin, antipain, and N-tosyl-l-lysine chloromethyl ketone. The inhibitor study suggests that the enzyme belongs to the class of the sulfhydryl proteases.

Original languageEnglish
Pages (from-to)358-363
Number of pages6
JournalArchives of Biochemistry and Biophysics
Volume250
Issue number2
DOIs
Publication statusPublished - Nov 1 1986

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Hydrolases
Zea mays
Purification
Arginine
Enzymes
Hydrophobic chromatography
Antipain
Sulfhydryl Reagents
Enzyme activity
Ion Exchange Chromatography
Ion exchange
Proteins
Peptide Hydrolases
Gels
Molecular weight
Gel Chromatography
Recovery
Molecular Weight
Substrates

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Purification and characterization of benzoyl-l-arginine p-nitroanilide hydrolase from etiolated leaves of Zea mays. / Doi, Michio; Shioi, Yuzo; Sasa, Tsutomu.

In: Archives of Biochemistry and Biophysics, Vol. 250, No. 2, 01.11.1986, p. 358-363.

Research output: Contribution to journalArticle

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