Partial purification and characterization of acylester hydrolase from Lupinus mutabilis
Anduaga, Walter ; Svensson, Ingemar LU ; Adlercreutz, Patrick LU
and Mattiasson, Bo
LU
(1999)
In JAOCS, Journal of the American Oil Chemists' Society
76(10).
p.1157-1162
- Abstract
An alkaline acylester hydrolase was partially purified from germinated seeds of Lupinus mutabilis. Hydrolytic activity was absent in the crude extract of ungerminated lupine seed, but it increased and peaked at the fourth day in the germinating seedling. The purification scheme involved homogenization, centrifugation, acetone precipitation, anion exchange chromatography, pH precipitation, and hydrophobic interaction chromatography. The acylester hydrolase was purified 126-fold, and the overall activity yield was 10%. The molecular weight estimated by sodium dodecylsulfate-polyacrylamide gel electrophoresis was 60 kDa. The enzyme had an isoelectric point of 6.2 and showed maximal activity at pH 8.0. The enzyme showed good stability... (More)
An alkaline acylester hydrolase was partially purified from germinated seeds of Lupinus mutabilis. Hydrolytic activity was absent in the crude extract of ungerminated lupine seed, but it increased and peaked at the fourth day in the germinating seedling. The purification scheme involved homogenization, centrifugation, acetone precipitation, anion exchange chromatography, pH precipitation, and hydrophobic interaction chromatography. The acylester hydrolase was purified 126-fold, and the overall activity yield was 10%. The molecular weight estimated by sodium dodecylsulfate-polyacrylamide gel electrophoresis was 60 kDa. The enzyme had an isoelectric point of 6.2 and showed maximal activity at pH 8.0. The enzyme showed good stability between pH 5.0 and 9.0. In the pH range 7.0-7.5, enzyme precipitation was observed. The enzyme was stable from 0 to 25°C for 5 h and at 45°C lost 50% of its activity in the same period of time. At higher temperatures, the enzyme showed low thermal stability. However, the highest initial activity was found to be at 45°C. Nonionic surfactants and cholic acid enhanced the activity of the enzyme. The activity was reduced by the addition of toluene and isooctane and increased by the addition of diethyl ether, acetonitrile, methanol, and pyridine. The activity was reduced by 37% in the presence of 1 mM Cu2+ ions. The enzyme-hydrolyzed triolein showing no positional specificity.
(Less)
- author
- Anduaga, Walter
; Svensson, Ingemar
LU
; Adlercreutz, Patrick
LU
and Mattiasson, Bo
LU
- organization
- publishing date
- 1999-10
- type
- Contribution to journal
- publication status
- published
- subject
- in
- JAOCS, Journal of the American Oil Chemists' Society
- volume
- 76
- issue
- 10
- pages
- 6 pages
- publisher
- The American Oil Chemists' Society
- external identifiers
-
- scopus:0033351127
- ISSN
- 0003-021X
- DOI
- 10.1007/s11746-999-0089-0
- language
- English
- LU publication?
- yes
- id
- d913adea-3f90-4ebd-a6f8-3a663bec2371
- date added to LUP
- 2019-06-20 15:34:59
- date last changed
- 2022-01-31 22:13:21
@article{d913adea-3f90-4ebd-a6f8-3a663bec2371,
abstract = {{<p>An alkaline acylester hydrolase was partially purified from germinated seeds of Lupinus mutabilis. Hydrolytic activity was absent in the crude extract of ungerminated lupine seed, but it increased and peaked at the fourth day in the germinating seedling. The purification scheme involved homogenization, centrifugation, acetone precipitation, anion exchange chromatography, pH precipitation, and hydrophobic interaction chromatography. The acylester hydrolase was purified 126-fold, and the overall activity yield was 10%. The molecular weight estimated by sodium dodecylsulfate-polyacrylamide gel electrophoresis was 60 kDa. The enzyme had an isoelectric point of 6.2 and showed maximal activity at pH 8.0. The enzyme showed good stability between pH 5.0 and 9.0. In the pH range 7.0-7.5, enzyme precipitation was observed. The enzyme was stable from 0 to 25°C for 5 h and at 45°C lost 50% of its activity in the same period of time. At higher temperatures, the enzyme showed low thermal stability. However, the highest initial activity was found to be at 45°C. Nonionic surfactants and cholic acid enhanced the activity of the enzyme. The activity was reduced by the addition of toluene and isooctane and increased by the addition of diethyl ether, acetonitrile, methanol, and pyridine. The activity was reduced by 37% in the presence of 1 mM Cu<sup>2+</sup> ions. The enzyme-hydrolyzed triolein showing no positional specificity.</p>}},
author = {{Anduaga, Walter and Svensson, Ingemar and Adlercreutz, Patrick and Mattiasson, Bo}},
issn = {{0003-021X}},
language = {{eng}},
number = {{10}},
pages = {{1157--1162}},
publisher = {{The American Oil Chemists' Society}},
series = {{JAOCS, Journal of the American Oil Chemists' Society}},
title = {{Partial purification and characterization of acylester hydrolase from <i>Lupinus mutabilis</i>}},
url = {{http://dx.doi.org/10.1007/s11746-999-0089-0}},
doi = {{10.1007/s11746-999-0089-0}},
volume = {{76}},
year = {{1999}},
}