Collagenase produced from Aspergillus sp. (UCP 1276) using chicken feather industrial residue.
An extracellular collagenolytic serine protease was purified from Aspergillus sp., isolated from the Caatinga biome in northeast Brazil by a two-step chromatographic procedure, using an anion-exchanger and gel filtration. The enzyme was produced by submerged fermentation of feather residue as a substrate. The purified collagenase showed a 2.09-fold increase in specific activity and 22.85% yield. The enzyme was a monomeric protein with a molecular mass of 28.7 kDa, estimated by an SDS-PAGE and AKTA system. The optimum temperature and pH for enzyme activity were around 40°C and pH 8.0, respectively. The enzyme was strongly inhibited by phenyl-methylsulfonyl fluoride, a serine protease inhibitor, and was thermostable until 65°C for 1 h. We then evaluated the enzyme's potential for degradation of Type I and Type V collagens for producing peptides with antifungal activity. Our results revealed that the cleavage of Type V collagen yielded more effective peptides than Type I, inhibiting growth of Aspergillus terreus, Aspergillus japonicus and Aspergillus parasiticus. Both groups of peptides (Type I and Type V) were identified by SDS-PAGE. To conclude, the thermostable collagenase we purified in this study has various potentially useful applications in the fields of biochemistry, biotechnology and biomedical sciences.
Department of Animal Morphology and Physiology, Rural Federal University of Pernambuco - UFRPE, Recife, PE, Brazil.|Department of Biotechnology, Laboratory of Imunopathology Keizo Asami, Federal University of Pernambuco, Recife, PE, Brazil.
Catarina Michelle Oliveira
CM
Ferreira
Department of Animal Morphology and Physiology, Rural Federal University of Pernambuco - UFRPE, Recife, PE, Brazil.
Patyanne Carvalho
PC
Correia
Department of Animal Morphology and Physiology, Rural Federal University of Pernambuco - UFRPE, Recife, PE, Brazil.
Romero Marcos Pedrosa
RM
Brandão-Costa
Department of Animal Morphology and Physiology, Rural Federal University of Pernambuco - UFRPE, Recife, PE, Brazil.
Wendell Wagner Campos
WW
Albuquerque
Department of Animal Morphology and Physiology, Rural Federal University of Pernambuco - UFRPE, Recife, PE, Brazil.
Tatiana Pereira Shin
TP
Lin Liu
Catholic University of Pernambuco, Recife, PE, Brazil.
Galba Maria
GM
Campos-Takaki
Department of Animal Morphology and Physiology, Rural Federal University of Pernambuco - UFRPE, Recife, PE, Brazil.
Ana Lúcia Figueiredo
AL
Porto
D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D001230:Aspergillus / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods*; D002645:Chickens; D017364:Collagenases / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D004795:Enzyme Stability; D005241:Feathers / Q000378:metabolism*; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology; D008970:Molecular Weight; D010446:Peptide Fragments / Q000494:pharmacology; D013696:Temperature; D014361:Trypsin Inhibitors / Q000494:pharmacology; D014866:Waste Products
D000935:Antifungal Agents; D061965:Matrix Metalloproteinase Inhibitors; D010446:Peptide Fragments; D014361:Trypsin Inhibitors; D014866:Waste Products; D017364:Collagenases
caatinga; collagenase; feather residue; proteases; purification