Two novel S1 peptidases from Amycolatopsis keratinophila subsp. keratinophila D2T degrading keratinous slaughterhouse by-products

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  • Roall Espersen
  • Francesco C. Falco
  • Per Hägglund
  • Krist V. Gernaey
  • Anna E. Lantz
  • Birte Svensson

Two proteases, named C- and T-like proteases, respectively, were purified from the culture supernatant of Amycolatopsis keratinophila subsp. keratinophila D2T grown on a keratinous slaughterhouse by-product of pig bristles and nails as sole nitrogen and carbon source. The two proteases belong to peptidase family S1 as identified by mass spectrometric peptide mapping, have low mutual sequence identity (25.8%) and differ in substrate specificity. T-like protease showed maximum activity at 40 °C and pH 8–9, and C-like protease at 60 °C and pH 8–10. Peptides released from the keratinous by-product were identified by mass spectrometry and indicated P1 specificity for arginine and lysine of T-like and alanine, valine and isoleucine of C-like protease as also supported by the activity of the two proteases towards synthetic peptide and amino acid substrates. The specific activities of the C- and T-like proteases and proteinase K on keratin azure and azokeratin were comparable. However, C- and T-like proteases showed 5–10-fold higher keratin/casein (K/C) activity ratios than that of another S1 and two keratin-degrading S8 peptidases used for comparison. The findings support that the range of peptidase families considered to contain keratinases should be expanded to include S1 peptidases. Furthermore, the results indicated the quite thermostable C-like protease to be a promising candidate for use in industrial degradation of keratinous slaughterhouse by-products.

Original languageEnglish
JournalApplied Microbiology and Biotechnology
Issue number6
Pages (from-to)2513-2522
Number of pages10
Publication statusPublished - 2020
Externally publishedYes

    Research areas

  • Amycolatopsis keratinophila, Keratin:casein activity ratio, Keratinase, S1 peptidase, Slaughterhouse by-product, Thermostability

ID: 240156966