title: Coupled action of γ-glutamyl transpeptidase-glutathione and keratinase effectively degrades feather keratin and surrogate prion protein, Sup 35NM.
abstract: Recombinant Escherichia coli HB101 harboring keratinase rKP2 from Pseudomonas aeruginosa KS-1 degraded 2% chicken feather in LB-Amp medium in 24h. SEM analysis and detailed studies revealed that bacterial colonization of feather was a pre-requisite for degradation of feather by keratinase. The mechanism of sulfitolysis revealed involvement of free cystinyl group as a source of redox during colonization as DTNB inhibited feather degradation by rKP2. Involvement of GGT-GSH system in contribution of free cystinyl group for redox was established by using GGT knockout recombinant E. coli strain that failed to degrade feather inspite of successful colonization and keratinase production. Short term experiments further confirmed enhanced protein release from feather keratin in presence of GGT-GSH redox. In the presence of similar redox, rKP2 also degraded surrogate prion protein, Sup 35NM in 15 min at 37°C, pH 7.0.
authors: Department of Microbiology, University of Delhi, South Campus, New Delhi 110021, India.
authors: Richa
authors: R
authors: Sharma
authors: Rani
authors: R
authors: Gupta
mesh_terms: D000818:Animals; D002645:Chickens; D015169:Colony Count, Microbial; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D005241:Feathers / Q000378:metabolism* / Q000382:microbiology / Q000648:ultrastructure; D005978:Glutathione / Q000494:pharmacology*; D007633:Keratins / Q000378:metabolism*; D010084:Oxidation-Reduction / Q000187:drug effects; D010447:Peptide Hydrolases / Q000494:pharmacology*; D010454:Peptide Termination Factors / Q000378:metabolism*; D011328:Prions / Q000378:metabolism; D059748:Proteolysis / Q000187:drug effects*; D011550:Pseudomonas aeruginosa / Q000201:enzymology; D011994:Recombinant Proteins / Q000494:pharmacology; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D013447:Sulfites / Q000378:metabolism; D005723:gamma-Glutamyltransferase / Q000494:pharmacology*
chemical_list: D010454:Peptide Termination Factors; D011328:Prions; D011994:Recombinant Proteins; C068068:SUP35 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D013447:Sulfites; D007633:Keratins; D005723:gamma-Glutamyltransferase; D010447:Peptide Hydrolases; C035855:keratinase; D005978:Glutathione