title: Identification of two new keratinolytic proteases from a Bacillus pumilus strain using protein analysis and gene sequencing.
abstract: The Bacillus strain (CCUG 66887) has a high capacity to excrete keratinase with the ability to degrade both alpha- and beta keratin. In this study we aimed to show the characteristics of the keratinolytic protease and to identify its gene by using liquid chromatography-electrospray ionization tandem mass spectrometry methods (nanoHPLC-ESI-MS/MS) followed by Mascot data base search. The results showed that the enzyme in fact consists of two different keratinases, both with a molecular mass of 38 kDa. Further, DNA sequencing generated the open reading frame (ORF) of one of the genes (Ker1), and de novo genome sequencing identified the ORF of the second gene (Ker2). The two keratinase genes contain 1153 base pairs each and have a gene similarity of 67 %. In addition, the Bacillus strain was classified as Bacillus pumilus and its genes were annotated in the GeneBank at NCBI (accession: CP011109.1). Amino acid sequences alignment with known B. pumilus proteases indicated that the two keratinases of B. pumilus strain C4 are subtilisin-like serine proteases belonging to the Protease S8 family. Taken together, these result suggest the two keratinases as promising candidates for enzymatic processing of keratinous wastes in waste refinery.
authors.affiliation: Department of Biology, Faculty of Sciences of Nature and Life, Mostaganem University, Mostaganem, Algeria.|Swedish Centre for Resource Recovery, University of Borås, Borås, Sweden.
authors.forename: Soltana
authors.initials: S
authors.lastname: Fellahi
authors.affiliation: Department of Biology, Faculty of Sciences of Nature and Life, Mostaganem University, Mostaganem, Algeria.
authors.forename: Abdelwaheb
authors.initials: A
authors.lastname: Chibani
authors.affiliation: Swedish Centre for Resource Recovery, University of Borås, Borås, Sweden. elisabeth.feuk@hb.se.
authors.forename: Elisabeth
authors.identifier: http://orcid.org/0000-0003-2029-7211
authors.initials: E
authors.lastname: Feuk-Lagerstedt
authors.affiliation: Swedish Centre for Resource Recovery, University of Borås, Borås, Sweden.
authors.forename: Mohammad J
authors.initials: MJ
authors.lastname: Taherzadeh
keywords: Bacillus pumilus; DNA sequencing; Keratinase; NanoHPLC–ESI–MS/MS; α-Keratin; β-Keratin