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http://dx.doi.org/10.1021/ac026136s

has been cited by the following article:

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

  AUTHORS: Nouani Abdelouahab, Belhamiche Nabila, Slamani Roza, Belbraouet Slimane, Dako Etienne, Audet Pascal, Bellal Mohand Mouloud

  KEYWORDS: Protease, Molecular Weight, Molecular Exclusion, Electrophoresis, Proteomics

  JOURNAL NAME: Food and Nutrition Sciences, Vol.6 No.3, March 13, 2015

  ABSTRACT: 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.