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Updated: Jul 18, 2026
Purification of Hsp104, a Protein Disaggregase
Published on: September 30, 2011
Purification and characterization of a keratinolytic metalloprotease from Chryseobacterium sp. kr6
Alessandro Riffel1, Adriano Brandelli, Cláudia de M Bellato
1EMBRAPA (Empresa Brasileira de Pesquisa Agropecuária) Tabuleiros Costeiros - UEP Rio Largo, caixa postal 2013, 57061-970 Maceió/AL, Brazil. ariffel@esalq.usp.br
Abstract:
The Chryseobacterium sp. kr6 strain has been described as a highly keratinolytic bacterium showing effective feather-degrading and de-hairing activities. A keratinase Q1 enzyme was purified from Chryseobacterium sp. kr6 culture by Phenyl Sepharose and Superose 12HR chromatography. This enzyme showed a specific activity of 967U/mg for keratin azure. Electrophoresis under denaturing conditions showed a monomeric protein with approximately 64kDa. The enzyme showed pH and temperature optima of 8.5 and 50 degrees C, respectively. The inhibitory effect of EDTA, EGTA and 1,10-phenanthroline characterized Q1 enzyme as a Zn-metalloprotease. Its activity was increased by three-fold in the presence of Ca(2+). ESI-MS/MS analysis of peptides generated from a tryptic digestion revealed sequence homology which may characterize the Q1 keratinase as a member of the M14 metalloprotease family, with a consensus glycosylation region similar to proteins from Chryseobacerium meningosepticum.
