Biodegradation of feather wastes

1. Isolation, Screening and Enzyme
Production of Potent Keratinolytic
Bacteria from Feather Wastes and
its Application
Bandana Subedi, Prabin Shakya And Amrit Acharya
Microbiological Research Organization Nepal
(MiRON)

2. Introduction
Solid waste management
–Discarded in open places
–Burning/incineration
–Compost
–Landfill site
–Not well managed

3. Keratin
• Fibrous structural proteins
• Recalcitrant
• Insoluble
• Alpha helix and beta sheets
• Sole carbon, nitrogen and sulphur source
• Feathers, hair, hoofs etc
• Mostly consists of valine, leucine, serine,
proline, aspartic and glutamic acid

4. Protofilament

5. Keratinase enzyme
• Proteolytic endopeptidase enzymes
• Produced by some insects and microbes
• Mostly serine protease and metalloprotease
• Attack on the disulfide (-S-S-) bond
• Monomer
• Molecular mass range from 20 kDa - 60 kDa
• 410-480 residues
• Encoded by kerA gene

6. Keratinase enzyme

7. Keratinolytic organisms
• Bacteria
– Bacillus spp.
– Arthrobacter spp.
• Actinomycetes
– Streptomyces spp.
– Nocardia spp.
• Fungi
– Microsporum spp.
– Trychophyton spp.
– Doratomyces microsporum
– Aspergillus fumigatus
– Aspergillus flavus

8. Objectives
• Isolation and screening of Keratinolytic
Bacteria
• Keratinase enzyme production
• Partial Purification of produced enzyme
• Activity of Keratinase enzyme in different
substrates

9. Methodology
• Sample: Soil, feather, Hair and other keratin rich
wastes
• Screening and Isolation of Potent isolates
– Heat treatment
– Serial dilution
– Plating in selective media
• Identification
– Morphological Characteristics
– Colonial Characteristics
– biochemical tests

10. Methodology….contd.
• Enzyme production
– batch fermentation process
– Feather/Keratin culture broth in
shaking in controlled environment
shaker
• Enzyme Purification
– Centrifugation
– Ammonium Sulphate Precipitation

11. Methodology….contd.
• Enzyme activity
– Hydrolysis of Keratin or other Keratin rich substrates
• Determination of Amino acids Released
– Thin Layer chromatography / Paper Chromatography
• Lab Trial
– Enzyme or potent bacterial isolates
– Substrates: Keratin, Hair, Feather
– Batch Fermentation
• Statistical Analysis
• Report writing
• Report submission
• Presentation of findings
• Publication

12. Application
• Solid waste management
• Biofertilizers
• Biofeed
• Single cell protien
• Amino acids
• Biogas
• Treatment of prion disease, BSE (Bovine
spongiform encephalitis)

13. Probable findings
• Potent Keratinolytic bacteria will be
isolated.
• Keratinase enzyme will be produced
and will be partially purified.
• The activity of keratinolytic bacteria and
its enzyme on feather will be assessed.
• Amino acid released will be evaluated.
• Laboratory Trial of the keratin rich solid
wastes

15. Thank you

16. Questions???