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

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology

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