Keratinase

Keratinases are proteolytic enzymes that digest keratin.

History

They were initially classified as 'proteinases of unknown mechanism' by the Nomenculture Committee on the International Union of Biochemistry in 1978 with EC number 3.4.99 in 1983 (Owen et al., 1983). In the 1990s, they were defined as a serine proteases due to high sequence homology with alkaline protease, and their inhibition by serine protease inhibitors (Wang et al., 1995; Taha et al., 1998 and Bressollier et a1., 1999).

Function

Keratinases are produced only in the presence of keratin-containing substrate. They mainly attack the disulfide (-S-S-) bond of the keratin substrate (Bockel et al., 1995). Keratinase production in various microorganisms was reported on by a number of researchers. It was found that keratinase in fungi, Streptomyces and bacteria were produced in nearly at alkaline pH and almost thermophilic temperatures. These enzymes have a wide range of substrate specificity such as it can degrade other fibrous protein fibrin, elastin, collagen and other non fibrous protein like casein, bovine serum albumin gelatin etc. (Noval et al., 1959; Mukhapadhayay et al., 1989; Dozie et al., 1994; Lin et al., 1995; Letourneau et al., 1998; and Bressollier et al., 1999).

Distribution

At first Molyneux et al. (1959) attempted to isolate some bacteria that are able to degrade keratin. He isolated organism from the contents of experimentally induced dermoid cysts from mid lateral region of sheep. Examination of wool sample showed degraded wool with numerous corticle and cyticular cells. He found disruption of wool fiber in both in vivo and in vitro. He showed that the organisms belong to genus Bacillus and the organism was capable of attacking native wool protein. The same year Noval et al. (1959) published another article on enzymatic decomposition of native keratin by Streptomyces fradiae. They showed extracellular enzyme secreted by these bacteria capable of degrading the human hair in its native state.

Keratinolytic protein from keratinophilic fungi were reported by Yu et al. (1968), Asahi et al. (1985), and Willams et al. (1989). Mukhopadhay et al. (1989) reported keratinase production by Streptomyces sp. He isolated an inducible extracellular homogenous enzyme, which shows a 7.5-fold increases in its activity after DEAE cellulose column chromatography. The enzyme-activity was inhibited by reduced glutathione, PMSF and 2-¬Mercaptaethanol.

Williams et al. (1990) continued his work on enriched feather degrading culture and characterized the organism to its species level for the first time. The microorganisms were identified as Bacillus licheniformis. Lin et al. (1992) purified and characterized keratinase from feather degrading Bacillus licheniformis strain isolated by Williams et al. (1990) with the help of membrane ultra filtration and C-75 gel chromatography. He purified enzyme with 70-fold increased activity. SDS-PAGE analysis revealed that purified keratinase had a molecular weight of 33 kDa. Dozie et al. (1994) reported a thermostable, alkaline-active, keratinolytic proteinasefrom Chrysosporium keratinophylum which was able to solubilize keratin in lactose-mineral salt medium with DMSO. Optimum pH for the enzyme activity was 9 and optimum temperature was 90 °C. Wang et al. (1999) scaled up the fermentation condition of keratinase to a pilot scale fermentar. They optimized the fermentation condition to a level of 10-fold increase in enzyme production.

References

1. Hoq MM, Siddiquee KAZ, Kawasaki H and Seki T.2005. Keratinolytic Activity of Some Newly Isolated Bacillus Species. Journal of Biological Science 5(2): 193-200, ISSN 1727-3048.

2. Lin X., Lee CG, Casale SE and Shih JC.1992. Purification and characterization of a keratinase from a feather degrading Bacillus Licheniformis strain. J. Appl. Environ. AIlcrobiol. 58 : 3271-3275.

3.Molyneux GS.1959. The digestion of wool by a keratinolytic Bacillus. Aus. J.Biol. Sci, 12: 274-281.

4. Williams CM, Richtcr CS, Mackenzie JR and Shih JCI.1990. Isolation, identification and characterization of a feather-degrading bacterium. J. Appl. Environ. Microbiol. 56: 1509-1515.