Biochemical and proteomic characterization of the extracellular enzymatic preparate of Exiguobacterium undae, suitable for efficient animal glue removal
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In this work, we describe the preparation and characterization of a biopreparate for efficient and rapid animal glue removal. The biopreparate is based on the extracellular proteolytic enzymes of an Exiguobacterium undae environmental isolate. Liquid chromatography-mass spectrometry analysis showed that the biopreparate is predominantly composed of hydrolytic enzymes—proteases and peptidases, nucleases, peptide ABC transporter substrate-binding proteins, and a phosphatase. The two main proteins present are bacillolysin and a peptide ABC transporter substrate-binding protein. Inhibition and proteomic analyses of the biopreparate revealed that bacillolysin, a neutral metalloendopeptidase, is mainly responsible for its proteolytic activity. This biopreparate was able to satisfactorily remove two types of animal glue from different kinds of material surfaces. These results suggest that this biopreparate could serve as a potential new tool for the restoration of historical objects rather than living microorganisms.
KeywordsExiguobacterium undae Animal glue Biodegradation Bacillolysin Extracellular proteome Liquid chromatography-mass spectrometry
The authors thank Dr. Zdenek Voburka (Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences, Prague, Czech Republic) for N-terminal sequencing and Dr. Jacob A. Bauer for discussion and manuscript revision.
The work was financially supported by the grant APVV-15-0528 “Modified polymers from renewable resources and their degradation.” This contribution is also the result of the project ITMS-26240220010 in the frame of the support program Research and Development of the European Regional Development Fund. A Slovak patent application form, No. PP50012-2018, has been applied for the E. undae biopreparate.
Compliance with ethical standards
This article does not contain any studies with human participants or animals performed by any of the authors.
Conflict of interest
The authors declare that they have no conflict of interest.
- Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, Bairoch A (2005) Protein identification and analysis tools on the ExPASy server. (In) J M. Walker (ed): The proteomics protocols handbook, Humana Press, pp. 571–607Google Scholar
- Harth L, Krah U, Linke D, Dunkel A, Hofmann T, Berger RG (2016) Salt taste enhancing l-arginyl dipeptides from casein and lysozyme released by peptidases of Basidiomycota. J Agric Food Chem https://doi.org/10.1021/acs.jafc.6b02716, in press
- Michalski A, Damoc E, Lange O, Denisov E, Nolting D, Müller M, Viner R, Schwartz J, Remes P, Belford M, Dunyach JJ, Cox J, Horning S, Mann M, Makarov A (2012) Ultra high resolution linear ion trap Orbitrap mass spectrometer (Orbitrap Elite) facilitates top down LC MS/MS and versatile peptide fragmentation modes. Mol Cell Proteomics 11:O111.013698CrossRefPubMedGoogle Scholar
- Sarmiento A, Pérez-Alonso M, Olivares M, Castro K, Martínez-Arkarazo I, Fernández LA, Madariaga JM (2011) Classification and identification of organic binding media in artworks by means of Fourier-transform infrared spectroscopy and principal component analysis. Anal Bioanal Chem 399:3601–3611CrossRefPubMedGoogle Scholar
- Selim S, Hassan S, Hagagy N, Kraková L, Grivalský T, Pangallo D (2017) Assessment of microbial diversity in Saudi springs by culture-dependent and culture-independent methods. Geomicrobiol J 34:443–453Google Scholar
- Ward GWR (2008) Materials and techniques in art. Oxford University Press, Editor: Ward GWRGoogle Scholar
- Wei S, Schreiner M, Rosenberg E, Guo H, Ma Q (2011) Identification of the binding media in Tang Dynasty Chinese wall paintings by using Py-GC/MS and GC/MS techniques. Int J Conserv Sci 2:77–88Google Scholar