Studies on Porphobilinogen-Deaminase from Saccharomyces cerevisiae
Porphobilinogen-deaminase from Saccharomyces cerevisiae has been isolated and partially purified 80- and 230-fold in the absence or presence of phenylmethylsulphonyl fluoride, re-spectively. Some properties of the isolated enzyme were studied. Porphyrin formation was linear with time and protein con...
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1991
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09395075_v46_n11-12_p1017_CorreaGarcia http://hdl.handle.net/20.500.12110/paper_09395075_v46_n11-12_p1017_CorreaGarcia |
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paper:paper_09395075_v46_n11-12_p1017_CorreaGarcia2023-06-08T15:53:26Z Studies on Porphobilinogen-Deaminase from Saccharomyces cerevisiae Porphobilinogen Porphobilinogen-Deaminase Porphyrin Biosynthesis Saccharomyces cerevisiae Yeast benzylsulfonyl fluoride porphobilinogen deaminase article chemistry enzymology gel chromatography isolation and purification kinetics metabolism methodology molecular weight pH Saccharomyces cerevisiae Chromatography, Gel Hydrogen-Ion Concentration Hydroxymethylbilane Synthase Kinetics Molecular Weight Phenylmethylsulfonyl Fluoride Saccharomyces cerevisiae Support, Non-U.S. Gov't Porphobilinogen-deaminase from Saccharomyces cerevisiae has been isolated and partially purified 80- and 230-fold in the absence or presence of phenylmethylsulphonyl fluoride, re-spectively. Some properties of the isolated enzyme were studied. Porphyrin formation was linear with time and protein concentration. Optimum pH was about 7.5-7.8. Molecular mass of the protein was 30,000 ± 3000 Dalton when the enzyme was purified in the presence of phenylmethyl-sulphonyl fluoride. A less active and unstable 20,000 Da molecular mass species was obtained when purification was performed in the absence of the protease inhibitor. Porphobilinogen-deaminase exhibited classical Michaelis-Menten kinetics. The apparent Km for uroporphyrinogen formation was 19 μM; Vmaxwas 3.6 nmol uroporphyrin/h and the Hill coefficient was n = 1. Also the action of several reagents on the activity was studied. Protective thiol agents had no effect. Heavy metals inhibited both porphyrin formation and porphobilinogen consumption, but known sulphydryl inactivating chemicals inhibit the former without modifying the latter. Ammonium ions had no effect on the activity while hydroxylamine completely inhibited both porphyrin formation and porphobilinogen consumption. © 1991, Verlag der Zeitschrift für Naturforschung. All rights reserved. 1991 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09395075_v46_n11-12_p1017_CorreaGarcia http://hdl.handle.net/20.500.12110/paper_09395075_v46_n11-12_p1017_CorreaGarcia |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Porphobilinogen Porphobilinogen-Deaminase Porphyrin Biosynthesis Saccharomyces cerevisiae Yeast benzylsulfonyl fluoride porphobilinogen deaminase article chemistry enzymology gel chromatography isolation and purification kinetics metabolism methodology molecular weight pH Saccharomyces cerevisiae Chromatography, Gel Hydrogen-Ion Concentration Hydroxymethylbilane Synthase Kinetics Molecular Weight Phenylmethylsulfonyl Fluoride Saccharomyces cerevisiae Support, Non-U.S. Gov't |
| spellingShingle |
Porphobilinogen Porphobilinogen-Deaminase Porphyrin Biosynthesis Saccharomyces cerevisiae Yeast benzylsulfonyl fluoride porphobilinogen deaminase article chemistry enzymology gel chromatography isolation and purification kinetics metabolism methodology molecular weight pH Saccharomyces cerevisiae Chromatography, Gel Hydrogen-Ion Concentration Hydroxymethylbilane Synthase Kinetics Molecular Weight Phenylmethylsulfonyl Fluoride Saccharomyces cerevisiae Support, Non-U.S. Gov't Studies on Porphobilinogen-Deaminase from Saccharomyces cerevisiae |
| topic_facet |
Porphobilinogen Porphobilinogen-Deaminase Porphyrin Biosynthesis Saccharomyces cerevisiae Yeast benzylsulfonyl fluoride porphobilinogen deaminase article chemistry enzymology gel chromatography isolation and purification kinetics metabolism methodology molecular weight pH Saccharomyces cerevisiae Chromatography, Gel Hydrogen-Ion Concentration Hydroxymethylbilane Synthase Kinetics Molecular Weight Phenylmethylsulfonyl Fluoride Saccharomyces cerevisiae Support, Non-U.S. Gov't |
| description |
Porphobilinogen-deaminase from Saccharomyces cerevisiae has been isolated and partially purified 80- and 230-fold in the absence or presence of phenylmethylsulphonyl fluoride, re-spectively. Some properties of the isolated enzyme were studied. Porphyrin formation was linear with time and protein concentration. Optimum pH was about 7.5-7.8. Molecular mass of the protein was 30,000 ± 3000 Dalton when the enzyme was purified in the presence of phenylmethyl-sulphonyl fluoride. A less active and unstable 20,000 Da molecular mass species was obtained when purification was performed in the absence of the protease inhibitor. Porphobilinogen-deaminase exhibited classical Michaelis-Menten kinetics. The apparent Km for uroporphyrinogen formation was 19 μM; Vmaxwas 3.6 nmol uroporphyrin/h and the Hill coefficient was n = 1. Also the action of several reagents on the activity was studied. Protective thiol agents had no effect. Heavy metals inhibited both porphyrin formation and porphobilinogen consumption, but known sulphydryl inactivating chemicals inhibit the former without modifying the latter. Ammonium ions had no effect on the activity while hydroxylamine completely inhibited both porphyrin formation and porphobilinogen consumption. © 1991, Verlag der Zeitschrift für Naturforschung. All rights reserved. |
| title |
Studies on Porphobilinogen-Deaminase from Saccharomyces cerevisiae |
| title_short |
Studies on Porphobilinogen-Deaminase from Saccharomyces cerevisiae |
| title_full |
Studies on Porphobilinogen-Deaminase from Saccharomyces cerevisiae |
| title_fullStr |
Studies on Porphobilinogen-Deaminase from Saccharomyces cerevisiae |
| title_full_unstemmed |
Studies on Porphobilinogen-Deaminase from Saccharomyces cerevisiae |
| title_sort |
studies on porphobilinogen-deaminase from saccharomyces cerevisiae |
| publishDate |
1991 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09395075_v46_n11-12_p1017_CorreaGarcia http://hdl.handle.net/20.500.12110/paper_09395075_v46_n11-12_p1017_CorreaGarcia |
| _version_ |
1768546683438759936 |