Purification and Partial Characterization of a Thrombin-Like Enzyme (AH144) from Venom of Iranian Snake Agkistrodon Halys
The snake venom´s thrombin-like enzymes comprise a number of serine proteases, which are functionally and structurally related to thrombin. Purification and partial characterization of a thrombin-like enzyme from the venom of the Iranian snake, Agkistrodon halys, was the aim of this study. Purificat...
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Iranian Institute of Research and Development in Chemical Industries (IRDCI)-ACECR
2012-06-01
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doaj-09a4d1d7886d44e69ced5991101097e42020-11-25T03:23:24ZengIranian Institute of Research and Development in Chemical Industries (IRDCI)-ACECRIranian Journal of Chemistry & Chemical Engineering 1021-99861021-99862012-06-013121031095997Purification and Partial Characterization of a Thrombin-Like Enzyme (AH144) from Venom of Iranian Snake Agkistrodon HalysMohammad Ghorbanpour0Farzin Zokaee Ashtiani1Abbas Zare Mirakabadi2Hosein Zolfagharian3Chemical Engineering Department, Amirkabir University of Technology, Tehran, I.R. IRANChemical Engineering Department, Amirkabir University of Technology, Tehran, I.R. IRANVenomous Animals and Antivenin production Department, Razi Vaccine and Serum Research Institute, Karaj, I.R. IRANVenomous Animals and Antivenin production Department, Razi Vaccine and Serum Research Institute, Karaj, I.R. IRANThe snake venom´s thrombin-like enzymes comprise a number of serine proteases, which are functionally and structurally related to thrombin. Purification and partial characterization of a thrombin-like enzyme from the venom of the Iranian snake, Agkistrodon halys, was the aim of this study. Purification was carried out by a combination of variety of chromatographic methods that included: gel filtration on Sephadex G-50, ion-exchange chromatography on DEAE-Sepharose and HPLC with a C18 column. A trial for the purification of protease resulted in an enzyme with specific activity of 721.2 (μmol/min/mg), which was purified by 72.1 fold. The purified thrombin-like enzyme designated AH144 was found to have a molecular weight of approximately 30.5 kDa. This thrombin-like enzyme had the highest activity at 37 °C and pH 7.5. Enzyme activity increased as its concentration increased, and the purified enzyme did not have any effect on casein. AH144 demonstrated clotting and proteolytic activities in the presence of the human plasma and the synthetic substrate (BApNA), respectively. Data emphasized the possibility of AH144 for quantitative determination of fibrinogen.http://www.ijcce.ac.ir/article_5997_7d43e894afc9aad9040e0083363f5e8d.pdfiranian snake venomagkistrodon halysthrombin-like enzymepurificationcoagulant activity |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Mohammad Ghorbanpour Farzin Zokaee Ashtiani Abbas Zare Mirakabadi Hosein Zolfagharian |
spellingShingle |
Mohammad Ghorbanpour Farzin Zokaee Ashtiani Abbas Zare Mirakabadi Hosein Zolfagharian Purification and Partial Characterization of a Thrombin-Like Enzyme (AH144) from Venom of Iranian Snake Agkistrodon Halys Iranian Journal of Chemistry & Chemical Engineering iranian snake venom agkistrodon halys thrombin-like enzyme purification coagulant activity |
author_facet |
Mohammad Ghorbanpour Farzin Zokaee Ashtiani Abbas Zare Mirakabadi Hosein Zolfagharian |
author_sort |
Mohammad Ghorbanpour |
title |
Purification and Partial Characterization of a Thrombin-Like Enzyme (AH144) from Venom of Iranian Snake Agkistrodon Halys |
title_short |
Purification and Partial Characterization of a Thrombin-Like Enzyme (AH144) from Venom of Iranian Snake Agkistrodon Halys |
title_full |
Purification and Partial Characterization of a Thrombin-Like Enzyme (AH144) from Venom of Iranian Snake Agkistrodon Halys |
title_fullStr |
Purification and Partial Characterization of a Thrombin-Like Enzyme (AH144) from Venom of Iranian Snake Agkistrodon Halys |
title_full_unstemmed |
Purification and Partial Characterization of a Thrombin-Like Enzyme (AH144) from Venom of Iranian Snake Agkistrodon Halys |
title_sort |
purification and partial characterization of a thrombin-like enzyme (ah144) from venom of iranian snake agkistrodon halys |
publisher |
Iranian Institute of Research and Development in Chemical Industries (IRDCI)-ACECR |
series |
Iranian Journal of Chemistry & Chemical Engineering |
issn |
1021-9986 1021-9986 |
publishDate |
2012-06-01 |
description |
The snake venom´s thrombin-like enzymes comprise a number of serine proteases, which are functionally and structurally related to thrombin. Purification and partial characterization of a thrombin-like enzyme from the venom of the Iranian snake, Agkistrodon halys, was the aim of this study. Purification was carried out by a combination of variety of chromatographic methods that included: gel filtration on Sephadex G-50, ion-exchange chromatography on DEAE-Sepharose and HPLC with a C18 column. A trial for the purification of protease resulted in an enzyme with specific activity of 721.2 (μmol/min/mg), which was purified by 72.1 fold. The purified thrombin-like enzyme designated AH144 was found to have a molecular weight of approximately 30.5 kDa. This thrombin-like enzyme had the highest activity at 37 °C and pH 7.5. Enzyme activity increased as its concentration increased, and the purified enzyme did not have any effect on casein. AH144 demonstrated clotting and proteolytic activities in the presence of the human plasma and the synthetic substrate (BApNA), respectively. Data emphasized the possibility of AH144 for quantitative determination of fibrinogen. |
topic |
iranian snake venom agkistrodon halys thrombin-like enzyme purification coagulant activity |
url |
http://www.ijcce.ac.ir/article_5997_7d43e894afc9aad9040e0083363f5e8d.pdf |
work_keys_str_mv |
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