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Published October 2015 | Supplemental Material
Journal Article Open

Metaproteomic evidence of changes in protein expression following a change in electrode potential in a robust biocathode microbiome

Abstract

Microorganisms that respire electrodes may be exploited for biotechnology applications if key pathways for extracellular electron transfer (EET) can be identified and manipulated through bioengineering. To determine whether expression of proposed Biocathode-MCL EET proteins are changed by modulating electrode potential without disrupting the relative distribution of microbial constituents, metaproteomic and 16S rRNA gene expression analyses were performed after switching from an optimal to suboptimal potential based on an expected decrease in electrode respiration. Five hundred and seventy-nine unique proteins were identified across both potentials, the majority of which were assigned to three previously defined Biocathode-MCL metagenomic clusters: a Marinobacter sp., a member of the family Chromatiaceae, and a Labrenzia sp. Statistical analysis of spectral counts using the Fisher's exact test identified 16 proteins associated with the optimal potential, five of which are predicted electron transfer proteins. The majority of proteins associated with the suboptimal potential were involved in protein turnover/turnover, motility, and membrane transport. Unipept and 16S rRNA gene expression analyses indicated that the taxonomic profile of the microbiome did not change after 52 hours at the suboptimal potential. These findings show that protein expression is sensitive to the electrode potential without inducing shifts in community composition, a feature that may be exploited for engineering Biocathode-MCL.

Additional Information

© 2015 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. Received: 09-Dec-2014; Revised: 23-Jun-2015; Accepted: 05-Aug-2015. We thank the DoD High Performance Computing Modernization Program's (HPCMP) PETTT staff at the Naval Research Laboratory for assistance with software configuration. We thank Dr. Masaru Miyagi from Case Western Reserve University, Cleveland, OH for kindly sharing purified SET. We thank Dr. Robert W. Li, USDA, for initial sequencing and annotation of the Biocathode-MCL metagenome. We thank the PRIDE Team for assistance with processing submission of mass spectrometry data to the ProteomeXchange Consortium. This work was funded by the Office of Naval Research via U.S. NRL core funds, as well as under the following award numbers to S.S.-G.: N0001413WX20995, N0001414WX20485, N0001414WX20518, and N0001415WX00195. The opinions and assertions contained herein are those of the authors and are not to be construed as those of the U.S. Navy, military service at large, or U.S. Government.

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