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Open Life Sciences

formerly Central European Journal of Biology

Editor-in-Chief: Ratajczak, Mariusz

1 Issue per year


IMPACT FACTOR 2016 (Open Life Sciences): 0.448

CiteScore 2016: 1.02

SCImago Journal Rank (SJR) 2016: 0.329
Source Normalized Impact per Paper (SNIP) 2016: 0.621

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Online
ISSN
2391-5412
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Volume 9, Issue 7 (Jul 2014)

Issues

Identification of proteins involved in starch and polygalacturonic acid degradation using LC/MS

Raimonda Petkauskaite
  • Department of Microbiology and Biotechnology, Faculty of Natural Sciences, Vilnius University, LT-03101, Vilnius, Lithuania
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/ Dangiras Lukosius
  • Department of Microbiology and Biotechnology, Faculty of Natural Sciences, Vilnius University, LT-03101, Vilnius, Lithuania
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/ Janusz Dębski
  • Mass Spectrometry Laboratory, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 02-106, Warsaw, Poland
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/ Andrius Jasilionis
  • Department of Microbiology and Biotechnology, Faculty of Natural Sciences, Vilnius University, LT-03101, Vilnius, Lithuania
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/ Michał Dadlez
  • Mass Spectrometry Laboratory, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 02-106, Warsaw, Poland
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/ Ieva Kieraite
  • Department of Microbiology and Biotechnology, Faculty of Natural Sciences, Vilnius University, LT-03101, Vilnius, Lithuania
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/ Ana Timonina
  • Department of Microbiology and Biotechnology, Faculty of Natural Sciences, Vilnius University, LT-03101, Vilnius, Lithuania
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/ Nomeda Kuisiene
  • Department of Microbiology and Biotechnology, Faculty of Natural Sciences, Vilnius University, LT-03101, Vilnius, Lithuania
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Published Online: 2014-04-30 | DOI: https://doi.org/10.2478/s11535-014-0303-4

Abstract

Plant biomass in the form of cheap wastes, such as straw, corn stalks, wood chips, sawdust, bagasse, pomace, etc., is abundant throughout the world. To convert these wastes into the useful value-added compounds microbial enzymes are the preferred choice. In this paper, we identify enzymes involved in the degradation of starch and polygalacturonic acid using liquid chromatography/mass spectrometry based analysis. We analysed total protein from soil and compost samples. Extracellular proteins from enrichment cultures were analysed in parallel and used as controls in the sample preparation and identification of proteins. In general, both protein sequence coverage and the number of identified peptides were higher in the samples obtained from the enrichment cultures than from the total protein from soil and compost. The influence of the nature of gel (zymography vs. SDS/polyacrylamide) was negligible. Thus, starch and polygalacturonic acid degradation associated proteins can be directly excised from the zymograms without the need to align zymograms with the SDS/polyacrylamide gels. A range of starch and polygalacturonic acid degradation associated enzymes were identified in both total protein samples and extracellular proteins from the enrichment cultures. Our results show that proteins involved in starch and polygalacturonic acid degradation can be identified by liquid chromatography/mass spectrometry from the complex protein mixtures both with and without cultivation of microorganism

Keywords: Total protein; Liquid chromatography/mass spectrometry; Starch degradation; Polygalacturonic acid degradation; Polysaccharide degradation; Biomass wastes; Thermophilic

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About the article

Published Online: 2014-04-30

Published in Print: 2014-07-01


Citation Information: Open Life Sciences, ISSN (Online) 2391-5412, DOI: https://doi.org/10.2478/s11535-014-0303-4.

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