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Genome and secretome analyses provide insights into keratin decomposition by novel proteases from the non-pathogenic fungus Onygena corvina.

Huang Y, Busk PK, Herbst FA, Lange L - Appl. Microbiol. Biotechnol. (2015)

Bottom Line: The culture broth was fractionated by ion exchange chromatography to isolate active fractions with five novel proteases belonging to three protease families (S8, M28, and M3).Enzyme blends composed of three of these five proteases, one from each family, showed high degree of degradation of keratin in vitro.A blend of novel proteases, such as those we discovered, could possibly find a use for degrading keratinaceous wastes and provide proteins, peptides, and amino acids as valuable ingredients for animal feed.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry and Bioscience, Aalborg University Copenhagen, 2450, Copenhagen, SV, Denmark.

ABSTRACT
Poultry processing plants and slaughterhouses produce huge quantities of feathers and hair/bristle waste annually. These keratinaceous wastes are highly resistant to degradation. Onygena corvina, a non-pathogenic fungus, grows specifically on feathers, hooves, horn, and hair in nature. Hence, the proteases secreted by O. corvina are interesting in view of their potential relevance for industrial decomposition of keratinaceous wastes. We sequenced and assembled the genome of O. corvina and used a method called peptide pattern recognition to identify 73 different proteases. Comparative genome analysis of proteases in keratin-degrading and non-keratin-degrading fungi indicated that 18 putative secreted proteases from four protease families (M36, M35, M43, and S8) may be responsible for keratin decomposition. Twelve of the 18 predicted protease genes could be amplified from O. corvina grown on keratinaceous materials and were transformed into Pichia pastoris. One of the recombinant proteases belonging to the S8 family showed high keratin-degrading activity. Furthermore, 29 different proteases were identified by mass spectrometry in the culture broth of O. corvina grown on feathers and bristle. The culture broth was fractionated by ion exchange chromatography to isolate active fractions with five novel proteases belonging to three protease families (S8, M28, and M3). Enzyme blends composed of three of these five proteases, one from each family, showed high degree of degradation of keratin in vitro. A blend of novel proteases, such as those we discovered, could possibly find a use for degrading keratinaceous wastes and provide proteins, peptides, and amino acids as valuable ingredients for animal feed.

No MeSH data available.


Related in: MedlinePlus

Spectrum of protease families in O. corvina genome (a), with number of protease genes from each family indicated; mass spectrometry data (b) identifying protease genes found in O. corvina secretome when grown on chicken feathers or pig bristle
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Fig2: Spectrum of protease families in O. corvina genome (a), with number of protease genes from each family indicated; mass spectrometry data (b) identifying protease genes found in O. corvina secretome when grown on chicken feathers or pig bristle

Mentions: Next, we applied PPR to find protease genes in the O. corvina genome. The hits were confirmed by blast, and full-length ORFs were predicted by Augustus. This led to identification of 73 putative proteases (Table S2). Clan information was obtained based on the Merops database, and the families were classified using the conserved domains. Most of the proteases are metalloproteases (M1, M3, M12, M14, M18, M19, M20, M28, M35, M36, M43, and M49), serine proteases (S8, S9, S10, S28, S33, S49, and S53), and cysteine proteases (C15 and C56). A majority of proteases belong to the families S8, S33, and M28 (Fig. 2a). A signal peptide was predicted in 12 of the 13 S8 proteases and in most of M28 proteases, which suggests that they are secreted proteins, whereas no signal peptide was predicted in any of S33 family proteases.Fig. 2


Genome and secretome analyses provide insights into keratin decomposition by novel proteases from the non-pathogenic fungus Onygena corvina.

Huang Y, Busk PK, Herbst FA, Lange L - Appl. Microbiol. Biotechnol. (2015)

Spectrum of protease families in O. corvina genome (a), with number of protease genes from each family indicated; mass spectrometry data (b) identifying protease genes found in O. corvina secretome when grown on chicken feathers or pig bristle
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4628079&req=5

Fig2: Spectrum of protease families in O. corvina genome (a), with number of protease genes from each family indicated; mass spectrometry data (b) identifying protease genes found in O. corvina secretome when grown on chicken feathers or pig bristle
Mentions: Next, we applied PPR to find protease genes in the O. corvina genome. The hits were confirmed by blast, and full-length ORFs were predicted by Augustus. This led to identification of 73 putative proteases (Table S2). Clan information was obtained based on the Merops database, and the families were classified using the conserved domains. Most of the proteases are metalloproteases (M1, M3, M12, M14, M18, M19, M20, M28, M35, M36, M43, and M49), serine proteases (S8, S9, S10, S28, S33, S49, and S53), and cysteine proteases (C15 and C56). A majority of proteases belong to the families S8, S33, and M28 (Fig. 2a). A signal peptide was predicted in 12 of the 13 S8 proteases and in most of M28 proteases, which suggests that they are secreted proteins, whereas no signal peptide was predicted in any of S33 family proteases.Fig. 2

Bottom Line: The culture broth was fractionated by ion exchange chromatography to isolate active fractions with five novel proteases belonging to three protease families (S8, M28, and M3).Enzyme blends composed of three of these five proteases, one from each family, showed high degree of degradation of keratin in vitro.A blend of novel proteases, such as those we discovered, could possibly find a use for degrading keratinaceous wastes and provide proteins, peptides, and amino acids as valuable ingredients for animal feed.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry and Bioscience, Aalborg University Copenhagen, 2450, Copenhagen, SV, Denmark.

ABSTRACT
Poultry processing plants and slaughterhouses produce huge quantities of feathers and hair/bristle waste annually. These keratinaceous wastes are highly resistant to degradation. Onygena corvina, a non-pathogenic fungus, grows specifically on feathers, hooves, horn, and hair in nature. Hence, the proteases secreted by O. corvina are interesting in view of their potential relevance for industrial decomposition of keratinaceous wastes. We sequenced and assembled the genome of O. corvina and used a method called peptide pattern recognition to identify 73 different proteases. Comparative genome analysis of proteases in keratin-degrading and non-keratin-degrading fungi indicated that 18 putative secreted proteases from four protease families (M36, M35, M43, and S8) may be responsible for keratin decomposition. Twelve of the 18 predicted protease genes could be amplified from O. corvina grown on keratinaceous materials and were transformed into Pichia pastoris. One of the recombinant proteases belonging to the S8 family showed high keratin-degrading activity. Furthermore, 29 different proteases were identified by mass spectrometry in the culture broth of O. corvina grown on feathers and bristle. The culture broth was fractionated by ion exchange chromatography to isolate active fractions with five novel proteases belonging to three protease families (S8, M28, and M3). Enzyme blends composed of three of these five proteases, one from each family, showed high degree of degradation of keratin in vitro. A blend of novel proteases, such as those we discovered, could possibly find a use for degrading keratinaceous wastes and provide proteins, peptides, and amino acids as valuable ingredients for animal feed.

No MeSH data available.


Related in: MedlinePlus