Kytococcus sedentarius, the organism associated with pitted keratolysis, produces two keratin-degrading enzymes
- 1 November 2002
- journal article
- Published by Oxford University Press (OUP) in Journal of Applied Microbiology
- Vol. 93 (5), 810-816
- https://doi.org/10.1046/j.1365-2672.2002.01742.x
Abstract
To determine characteristics of the extracellular enzyme activity of Kytococcus sedentarius on human callus. A concentrate of a continuous culture supernatant fluid of K. sedentarius, which had callus-degrading activity, was subjected to a series of chromatographic purification procedures. The enzyme activity was found to be attributable to two proteases. These were capable of degrading both native callus and extracted keratin polypeptides and were purified to homogeneity, as shown by SDS-PAGE with silver staining. The enzymes P1 and P2 were 30 kDa and 50 kDa in size with isoelectric points of 4.6 and 2.7, respectively. The optimum conditions for callus-degrading activity were 40 degrees C, pH 7.1 for P1 and 50 degrees C, pH 7.5 for P2. P2 displayed increased activity in the presence of 800 mmol l(-1) NaCl and both enzymes were inhibited by PMSF (1 mmol(-1) Phenylmethylsulphoryl fluoride) and 1 mmol l(-1) EDTA. The main enzyme cleavage sites were Lys-Trp, Val-Lys, Gly-Asp and Asp-Arg, as determined after incubation of P1 and P2 with the beta-chain of insulin. K. sedentarius produces two extracellular enzymes that independently degrade natural, insoluble human callus. Both enzymes are serine proteases and have cleavage preference sites that are present in a range of human keratins. The identification, in K. sedentarius cultures, of two enzymes which can degrade human callus strengthens the hypothesis that this organism is responsible for the pitting in human epidermis observed in pitted keratolysis. These enzymes may be of commercial use in the biodegradation of a range of keratin polymers, biological washing powders and in the treatment of unwanted callus on human skin.Keywords
This publication has 11 references indexed in Scilit:
- A review: Potentials for biotechnological applications of keratin-degrading microorganisms and their enzymes for nutritional improvement of feathers and other keratins as livestock feed resourcesBioresource Technology, 1998
- Keratins and the SkinAnnual Review of Cell and Developmental Biology, 1995
- Recent Development in Poultry Waste Dizgestion and Feather Utilization—A Review ,Poultry Science, 1993
- The effect of dilution rate and pH on biomass and proteinase production by Micrococcus sedentarius grown in continuous cultureJournal of Applied Bacteriology, 1992
- Qualitative and quantitiative assay for detection of callus degrading activity by bacteriaLetters in Applied Microbiology, 1990
- A comparative study of the cutaneous microflora of normal feet with low and high levels of odourJournal of Applied Bacteriology, 1988
- Pitted keratolysis. The role of Micrococcus sedentariusArchives of Dermatology, 1987
- Modification of human prekeratin during epidermal differentiationBiochemical Journal, 1981
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- Characterization of Proteinases Excreted by Bacillus subtilis Marburg Strain during SporulationJournal of Applied Bacteriology, 1970