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 language | English |
|---|---|
| Pages (from-to) | 358-363 |
| Number of pages | 6 |
| Journal | Archives of Biochemistry and Biophysics |
| Volume | 250 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Nov 1 1986 |
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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 journal › Article
}
TY - JOUR
T1 - Purification and characterization of benzoyl-l-arginine p-nitroanilide hydrolase from etiolated leaves of Zea mays
AU - Doi, Michio
AU - Shioi, Yuzo
AU - Sasa, Tsutomu
PY - 1986/11/1
Y1 - 1986/11/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0022808964&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0022808964&partnerID=8YFLogxK
U2 - 10.1016/0003-9861(86)90737-X
DO - 10.1016/0003-9861(86)90737-X
M3 - Article
C2 - 3535677
AN - SCOPUS:0022808964
VL - 250
SP - 358
EP - 363
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
SN - 0003-9861
IS - 2
ER -