Keratinases are exciting proteolytic enzymes that display the capability to degrade the insoluble protein keratin. These enzymes are produced by diverse microorganisms belonging to the Eucarya, Bacteria, and Archea domains. Keratinases display a great diversity in their biochemical and biophysical properties. Most keratinases are optimally active at neutral to alkaline pH and 40-60 degrees Celsius, but examples of microbial keratinolysis at alkalophilic and thermophilic conditions have been well documented. Several keratinases have been associated to the subtilisin family of serine-type proteases by analysis of their protein sequences. Studies with specific substrates and inhibitors indicated that keratinases are often serine or metalloproteases with preference for hydrophobic and aromatic residues at the P1 position. Keratinolytic enzymes have several current and potential applications in agroindustrial, pharmaceutical, and biomedical fields. Their use in biomass conversion into biofuels may address the increasing concern on energy conservation and recycling.

A novel feather-degrading microorganism was isolated from poultry waste, producing a high keratinolytic activity when cultured on broth containing native feather. Complete feather degradation was achieved during cultivation. The bacterium presents potential use for biotechnological processes involving keratin hydrolysis. Chryseobacterium sp. strain kr6 was identified based on morphological and biochemical tests and 16S rRNA sequencing. The bacterium presented optimum growth at pH 8.0 and 30 degrees C; under these conditions, maximum feather-degrading activity was also achieved. Maximum keratinase production was reached at 25 degrees C, while concentration of soluble protein was similar at both 25 and 30 degrees C. Reduction of disulfide bridges was also observed, increasing with cultivation time. The keratinase of strain kr6 was active on azokeratin and azocasein as substrates, and presented optimum pH and temperature of 7.5 and 55 degrees C, respectively. The keratinase activity was inhibited by 1,10-phenanthroline, EDTA, Hg(2+), and Cu(2+) and stimulated by Ca(2+).

The aim of this study was to characterize keratinolytic bacteria isolated from feather waste. Four isolates were selected after growth on solid medium with feather meal as sole carbon and nitrogen source and screened for proteolytic activity on milk agar plates. Three isolates were Gram-negative (belonged to the genera Burkholderia, Chryseobacterium and Pseudomonas) and one was Gram-positive (Microbacterium sp.). These bacteria grew on diverse keratin wastes such as feather meal, raw feathers, chicken nails, hair and wool. Keratinase activity was detected during growth, but the complete degradation of these substrates was not always achieved. The proteolytic character of crude enzymes was assessed using azokeratin and azocasein as substrates. The keratinases were active on both substrates and were similar in keratin hydrolysis when compared with commercially available microbial peptidases. These novel keratinolytic isolates have potential biotechnological use in processes involving keratin hydrolysis.

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