Molecular Weight Determination of a Protease Extracted from iMucor pusillus/i: Comparison Methods

Nouani Abdelouahab; Belhamiche Nabila; Slamani Roza; Belbraouet Slimane; Dako Etienne; Audet Pascal; Bellal Mohand Mouloud

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Molecular Weight Determination of a Protease Extracted from iMucor pusillus/i: Comparison Methods

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Mucor pepsin, a protease used in milk coagulation, is purified by ion-exchange and by molecular exclusion on Sephadex G100. The molecular weight (MW) is determined by polyacrylamide gel electrophoresis under denaturing conditions in presence of sodium dodecyl sulfate (SDS) and by molecular exclusion chromatography. Approximate evaluation of molecular mass was conducted by elution of known MW proteins (BSA: 67 kDa, pepsin: 35 kDa and trypsin: 23.8 kDa) on Sephadex G-100 under the same conditions as the experimental sample. The electrophoretic profile shows that the active fraction studied appears as a single homogeneous band (monomeric form). According to the curve calibration, the molecular mass of the coagulant fraction is about 48 kDa. For Mucor, the observed MW value seems to be enigmatic. However, this result is confirmed by a proteomic analysis with close MW values obtained using conventional techniques. The protease studied by the Scafold ver. Software 2.0 and the analysis of the protein similarities indicate a MW of 46 kDa and the protease sequence of 427 amino acids.

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《食品与营养科学（英文）》 |2015年第3期|348-354|共7页
作者
Nouani Abdelouahab; Belhamiche Nabila; Slamani Roza; Belbraouet Slimane; Dako Etienne; Audet Pascal; Bellal Mohand Mouloud;
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作者单位

Food Laboratory Technology;

Boumerdes University;

Boumerdes;

Algeria;

Laboratory of Food Technology;

Superior National School of Agronomy;

Algiers;

Algeria;

University of Bejaia;

Bejaia;

Algeria;

National Agricultural Research Institute;

Algiers;

Algeria;

éSANéF;

University of Moncton;

Moncton;

Canada;

Centre de Recherche sur les Aliments;

University of Moncton;

Moncton;

Canada;

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原文格式 PDF
正文语种 chi
中图分类肿瘤学;
关键词
Protease; Molecular; Weight; Molecular; Exclusion; Electrophoresis; Proteomics;

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Molecular Weight Determination of a Protease Extracted from iMucor pusillus/i: Comparison Methods

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