Molecular Weight Determination of a Protease Extracted from Mucor pusillus: Comparison Methods

Autor:	Bellal Mohand Mouloud, Belbraouet Slimane, Belhamiche Nabila, Slamani Roza, Nouani Abdelouahab, Dako Etienne, Audet Pascal
Rok vydání:	2015
Předmět:	Mucor Chromatography Protease Molecular mass biology Elution medicine.medical_treatment biology.organism_classification Trypsin chemistry.chemical_compound chemistry Biochemistry Sephadex medicine Sodium dodecyl sulfate Polyacrylamide gel electrophoresis medicine.drug
Zdroj:	Food and Nutrition Sciences. :348-354
ISSN:	2157-9458 2157-944X
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::6bb98ad10bee2b222b84efbfbb964f9f https://doi.org/10.4236/fns.2015.63035 Zobrazit plný text záznamu