microbial degradation of poultry feather

1. Topic :- Microbial degradation of poultry
feather biomass by KLEBSIELLA SP
. isolated
from poultry waste disposal site
Presented by :- Qurrat-ul-ain
Presented to :- Dr Pervez
Roll no :- 20204022-021

2. Introduction
 Enormous number of waste are generated through various food
industries among which meat industry produce a waste biomass
consisting noticeable quantities of organic residue such as feather
bones etc. . Chicken feather is the major by product- generated in
millions of tones from the commercial poultry processing which is
accumulating at higher rate (Zhao et al .2012).
 The disposal of chicken carcasses present significant environmental
,biological and financial problems for the poultry industry . This waste
having a large amount of protein such as keratin
 we foucused that some bacterial species like KLEBSIELA SP. As we
know that keratin is a insoluble protein . Through bacterial specie
keratinase enzyme produced to hydrolyze the keratin waste and utilize
nitrogen source for growth and development .

3. Hypothesis
 Keratinase enzyme can be used to hydrolyze poultry feather waste.

4. objectives
 using bacterial specie KLEBSIELLA SP. to hydrolyze the poultry feathers
 Analysis of enzyme activity and products of feather degradation.
 Using keratin hydrolysate as a bio fertilizer.

5. Material and methods :-
Chemicals and keratinous substrates:- Standard keratin, bovine serum albumin,
standard amino acids, di-sodium hydrogen orthophosphate (Na2HPO4), potassium di-
hydrogen orthophosphate (KH2PO4), ammonium chloride (NH4Cl), sodium chloride
(NaCl), magnesium sulphate (MgSO4), tri-chloro acetic acid (TCA), folin-phenol
reagent are collected through lab experiment .
 Chickens were first slaughter and the feathers were scalded from their body in an
automated machine at chicken slaughtering center.
 This obtained feather biomass was free of body parts, washed under tap water in
the mesh tray. Chicken feathers were then dried under sun light and stored for the
further experiments. Other keratinous substrates such as human hair (from barber
shop), sheep wool and silk cocoons were collected from local market of Sialkot.
Isolation of keratin utilizing microorganism
 The strains BTSUK was first screened for proteolytic
Potential of milk agar plate containing basal salt medium
and keratin powder (0.1%) as a sole source of carbon, nitrogen
and energy and cultures were maintained on the same medium
(Gurav and Jadhav, 2012).

6. Molecular identification of bacteria:- The identification of the strain was
performed by 16S rRNA gene sequence analysis .The obtained nucleotide
sequence was submitted to GenBank under accession number JX477370 This
bacterium was aligned with non redundant database present in NCBI using
BLASTn program and the homologous sequences of species obtained were
used for phylogenic analysis .
Phylogenetic position of sp. BTSUK within the genus Klebsiella and allied bacteria. The
branching pattern was generated by neighbor-joining method. The number of each branch
indicates the bootstrap values.

7. Feather degradation:- The feather degradation was carried out in sterilized liquid BSM
containing native
chicken feathers (1% w/v) inoculated with Klebsiella sp BTSUK and incubated on rotary (140
rpm) at 37 C.
Keratinase enzyme, soluble protein and amino acid analysis :-
The reaction mixture consisted of 1.0 ml crude extracellular enzyme diluted with tris HCL
buffer (pH 8.5) and 1.0 ml of 0.1% (w/v) standard keratin (dissolved in buffer) as a substrate
This mixture was incubated at 37 °C for 10 min and the reaction was stopped by adding 2.0ml
(0.4 mol l -1 ) trichloroacetic acid (TCA) and the supernatant after centrifugation was used to
Determine the enzyme activity at 280 nm (Gurav and Jadhav, 2012) .
Keratinase activity and percentage degradation on different
keratin substrates :-
The Klebsiella sp. BTSUK was tested to degrade keratin substrates like chicken feathers,
human hair, wool and silk. The BSM was supplemented with different keratinous substrates
(1.0% w/v) and incubated at 37°C on orbital shaker (140 rmp) and analyzed for keratinases
activity .

8. Possible
outcomes
Biodegradation of
feathers indicated
by pH meter due
to the presence of
simpler peptides.
Degradation of
wool, silk and hair
by keratinases.
Identification
bacterial strain
involved in
biodegradation.

9. Future Prospective
 Further research is needed on keratin waste that can serve as raw material in
biocompatible proteins, textile fibers, microcapsules, bioplastics, edible
materials, and production of rare amino acids like serine, serine and proline.
 Research on keratinases in order to use for further waste treatment.
 Use of keratinases in leather industries in order to treat animal skin before
turning into stuff is yet to be done.