Processing of poultry feathers by alkaline keratin hydrolyzing enzyme from Serratia sp. HPC 1383.
The present study describes the production and characterization of a feather hydrolyzing enzyme by Serratia sp. HPC 1383 isolated from tannery sludge, which was identified by the ability to form clear zones around colonies on milk agar plates. The proteolytic activity was expressed in terms of the micromoles of tyrosine released from substrate casein per ml per min (U/mL min). Induction of the inoculum with protein was essential to stimulate higher activity of the enzyme, with 0.03% feathermeal in the inoculum resulting in increased enzyme activity (45U/mL) that further increased to 90U/mL when 3d old inoculum was used. The highest enzyme activity, 130U/mL, was observed in the presence of 0.2% yeast extract. The optimum assay temperature and pH for the enzyme were found to be 60 degrees C and 10.0, respectively. The enzyme had a half-life of 10min at 60 degrees C, which improved slightly to 18min in presence of 1mM Ca(2+). Inhibition of the enzyme by phenylmethyl sulfonyl fluoride (PMSF) indicated that the enzyme was a serine protease. The enzyme was also partially inhibited (39%) by the reducing agent beta-mercaptoethanol and by divalent metal ions such as Zn(2+) (41% inhibition). However, Ca(2+) and Fe(2+) resulted in increases in enzyme activity of 15% and 26%, respectively. The kinetic constants of the keratinase were found to be 3.84 microM (K(m)) and 108.7 microM/mLmin (V(max)). These results suggest that this extracellular keratinase may be a useful alternative and eco-friendly route for handling the abundant amount of waste feathers or for applications in other industrial processes.
Khardenavis
Anshuman A
AA
National Environmental Engineering Research Institute, Nagpur 440 020, Maharashtra, India.
Kapley
Atya
A
Purohit
Hemant J
HJ
D000818:Animals; D002645:Chickens; D003470:Culture Media; D005241:Feathers / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007220:Industrial Waste / Q000032:analysis; D007633:Keratins / Q000378:metabolism; D007700:Kinetics; D008670:Metals / Q000378:metabolism; D010447:Peptide Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D012705:Serratia / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D013633:Tanning; D013696:Temperature
D003470:Culture Media; D007220:Industrial Waste; D008670:Metals; D007633:Keratins; D010447:Peptide Hydrolases