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Insects 2016, 7(4), 54; doi:10.3390/insects7040054

Label-Free Differential Proteomics and Quantification of Exoenzymes from Isolates of the Entomopathogenic Fungus Beauveria bassiana

1
Department of Molecular Biology, Aarhus University, Flakkebjerg, Forsøgsvej 1, 4200 Slagelse, Denmark
2
Department of Agroecology—Entomology and Plant Pathology, Aarhus University, Flakkebjerg, Forsøgsvej 1, 4200 Slagelse, Denmark
*
Author to whom correspondence should be addressed.
Academic Editor: Brian T. Forschler
Received: 25 April 2016 / Revised: 21 September 2016 / Accepted: 1 October 2016 / Published: 14 October 2016
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Abstract

Beauveria bassiana is an entomopathogenic fungus that grows both in vivo and in vitro. In vivo it can colonize live insect hosts, and tissue digestion occurs by secreted hydrolytic exoenzymes. It can also colonize dead insect tissue provided this is free from competing microorganisms. Depending on whether the host is alive or dead the expression (quality/quantity) of the exoenzymes may vary. We have grown several isolates of B. bassiana in shaking flasks for 120 h at 25 °C in order to evaluate the maximal exoenzyme production using two diet regimes. As sole carbon, nitrogen, and phosphate sources we used 1% shrimp chitin and either 0.5% w/v of dead intact American cockroach (Periplaneta americana) or their isolated cuticles. This is the first report of a differential proteomics of B. bassiana exoenzymes performed by label-free nano-LC MS/MS. Total proteolytic enzyme activity was mainly due to Pr1A or Pr1B depending on the isolate and the diet regime. The most differentially secreted enzymes were: the cuticle-degrading subtilisin Pr1A, GH13 alpha-glycosidase, glucan endo-1,3-beta-glucosidase, subtilisin-like proteinase Spm1, lipase 1, beta-1,3 exoglucanase, and endo-1,3-beta-glucosidase. Among the B. bassiana isolates analyzed, Bb 678 and Bb BG were the most active in Pr1A secretion. View Full-Text
Keywords: Beauveria bassiana; differential proteomics; Progenesis QI for proteomics; exoenzymes; subtilisin protease; chitinases; cockroach; cuticle-degrading serine proteinase Beauveria bassiana; differential proteomics; Progenesis QI for proteomics; exoenzymes; subtilisin protease; chitinases; cockroach; cuticle-degrading serine proteinase
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Dionisio, G.; Kryger, P.; Steenberg, T. Label-Free Differential Proteomics and Quantification of Exoenzymes from Isolates of the Entomopathogenic Fungus Beauveria bassiana. Insects 2016, 7, 54.

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