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Identification of two new keratinolytic proteases from a Bacillus pumilus strain using protein analysis and gene sequencing
University of Borås, Faculty of Textiles, Engineering and Business. (Resource Recovery)ORCID iD: 0000-0003-4887-2433
2016 (English)In: AMB Express, ISSN 2191-0855, E-ISSN 2191-0855, Vol. 6, no 1Article in journal (Refereed) Published
Abstract [en]

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.[on SciFinder (R)]

Place, publisher, year, edition, pages
2016. Vol. 6, no 1
Keyword [en]
bacillus pumilus, dna sequencing, keratinase, nanohplc-esi-ms/ms, α-keratin, β-keratin
National Category
Industrial Biotechnology
Research subject
Resource Recovery
Identifiers
URN: urn:nbn:se:hb:diva-10774DOI: 10.1186/s13568-016-0213-0ISI: 000380004700001ScopusID: 2-s2.0-84977651162OAI: oai:DiVA.org:hb-10774DiVA: diva2:1033365
Note

MEDLINE AN 2017159096(Journal; Article; (JOURNAL ARTICLE))

Available from: 2016-10-06 Created: 2016-09-27 Last updated: 2016-12-07Bibliographically approved

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CiteExportLink to record
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