Exploring bioactive peptides from bacterial secretomes using PepSAVI-MS: identification and characterization of Bac-21 from Enterococcus faecalis pPD1

Christine L. Kirkpatrick; Nicole C. Parsley; Tessa E. Bartges; Casey E. Wing; Sushma Kommineni; Christopher J. Kristich; Nita H. Salzman; Steven M. Patrie; Leslie M. Hicks

doi:10.1111/1751-7915.13299

Exploring bioactive peptides from bacterial secretomes using PepSAVI-MS: identification and characterization of Bac-21 from Enterococcus faecalis pPD1

Christine L. Kirkpatrick, Nicole C. Parsley, Tessa E. Bartges, Casey E. Wing, Sushma Kommineni, Christopher J. Kristich, Nita H. Salzman, Steven M. Patrie, Leslie M. Hicks^*

^*Corresponding author for this work

Chemistry

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

As current methods for antibiotic drug discovery are being outpaced by the rise of antimicrobial resistance, new methods and innovative technologies are necessary to replenish our dwindling arsenal of antimicrobial agents. To this end, we developed the PepSAVI-MS pipeline to expedite the search for natural product bioactive peptides. Herein we demonstrate expansion of PepSAVI-MS for the discovery of bacterial-sourced bioactive peptides through identification of the bacteriocin Bac-21 from Enterococcus faecalis pPD1. Minor pipeline modifications including implementation of bacteria-infused agar diffusion assays and optional digestion of peptide libraries highlight the versatility and wide adaptability of the PepSAVI-MS pipeline. Additionally, we have experimentally validated the primary protein sequence of the active, mature Bac-21 peptide for the first time and have confirmed its identity with respect to primary sequence and post-translational processing. Successful application of PepSAVI-MS to bacterial secretomes as demonstrated herein establishes proof-of-principle for use in novel microbial bioactive peptide discovery.

Original language	English (US)
Pages (from-to)	943-951
Number of pages	9
Journal	Microbial Biotechnology
Volume	11
Issue number	5
DOIs	https://doi.org/10.1111/1751-7915.13299
State	Published - Sep 2018

Funding

Received 25 March, 2018; revised 30 May, 2018; accepted 21 June, 2018. *For correspondence. E-mail: [email protected]; Tel. +1-919-843-6903; Fax +1-919-962-2388. Microbial Biotechnology (2018) 11(5), 943–951 doi:10.1111/1751-7915.13299 Funding Information This work was supported by NIH-NIGMS under award number 1R01GM125814 to L.M.H. and 1R01GM115739 to S.M.P. Financial support for C.L.K. was provided, in part, through the Eastman Summer Fellowship in Analytical Chemistry. N.C.P. acknowledges support from the NIH Molecular and Cellular Biophysics training grant (T32 GM008570) and T.E.B. acknowledges support from the National Science Foundation Graduate Research Fellowship Program (DGE-1650016). NSF MRI (CHE-1726291) supported the purchase of the Q-Exactive HF-X mass spectrometer, and we thank Dr. Brandie Ehrmann for training on the HF-X instrument. Funding Information This work was supported by NIH-NIGMS under award number 1R01GM125814 to L.M.H. and 1R01GM115739 to S.M.P. Financial support for C.L.K. was provided, in part, through the Eastman Summer Fellowship in Analytical Chemistry. N.C.P. acknowledges support from the NIH Molecular and Cellular Biophysics training grant (T32 GM008570) and T.E.B. acknowledges support from the National Science Foundation Graduate Research Fellowship Program (DGE-1650016). NSF MRI (CHE-1726291) supported the purchase of the Q-Exactive HF-X mass spectrometer, and we thank Dr. Brandie Ehrmann for training on the HF-X instrument. This work was supported by NIH-NIGMS under award number 1R01GM125814 to L.M.H. and 1R01GM115739 to S.M.P. Financial support for C.L.K. was provided, in part, through the Eastman Summer Fellowship in Analytical Chemistry. N.C.P. acknowledges support from the NIH Molecular and Cellular Biophysics training grant (T32 GM008570) and T.E.B. acknowledges support from the National Science Foundation Graduate Research Fellowship Program (DGE-1650016). NSF MRI (CHE-1726291) supported the purchase of the Q-Exactive HF-X mass spectrometer, and we thank Dr. Brandie Ehrmann for training on the HF-X instrument. We dedicate this manuscript to the Department of Chemistry at the University of North Carolina at Chapel Hill, in acknowledgement of its 200th anniversary, 1818?2018. This work was supported by NIH-NIGMS under award number 1R01GM125814 to L.M.H. and 1R01GM115739 to S.M.P. Financial support for C.L.K. was provided, in part, through the Eastman Summer Fellowship in Analytical Chemistry. N.C.P. acknowledges support from the NIH Molecular and Cellular Biophysics training grant (T32 GM008570) and T.E.B. acknowledges support from the National Science Foundation Graduate Research Fellowship Program (DGE-1650016). NSF MRI (CHE-1726291) supported the purchase of the Q-Exactive HF-X mass spectrometer, and we thank Dr. Brandie Ehrmann for training on the HF-X instrument. We dedicate this manuscript to the Department of Chemistry at the University of

ASJC Scopus subject areas

Biotechnology
Bioengineering
Biochemistry
Applied Microbiology and Biotechnology

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Kirkpatrick, C. L., Parsley, N. C., Bartges, T. E., Wing, C. E., Kommineni, S., Kristich, C. J., Salzman, N. H., Patrie, S. M., & Hicks, L. M. (2018). Exploring bioactive peptides from bacterial secretomes using PepSAVI-MS: identification and characterization of Bac-21 from Enterococcus faecalis pPD1. Microbial Biotechnology, 11(5), 943-951. https://doi.org/10.1111/1751-7915.13299

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title = "Exploring bioactive peptides from bacterial secretomes using PepSAVI-MS: identification and characterization of Bac-21 from Enterococcus faecalis pPD1",

abstract = "As current methods for antibiotic drug discovery are being outpaced by the rise of antimicrobial resistance, new methods and innovative technologies are necessary to replenish our dwindling arsenal of antimicrobial agents. To this end, we developed the PepSAVI-MS pipeline to expedite the search for natural product bioactive peptides. Herein we demonstrate expansion of PepSAVI-MS for the discovery of bacterial-sourced bioactive peptides through identification of the bacteriocin Bac-21 from Enterococcus faecalis pPD1. Minor pipeline modifications including implementation of bacteria-infused agar diffusion assays and optional digestion of peptide libraries highlight the versatility and wide adaptability of the PepSAVI-MS pipeline. Additionally, we have experimentally validated the primary protein sequence of the active, mature Bac-21 peptide for the first time and have confirmed its identity with respect to primary sequence and post-translational processing. Successful application of PepSAVI-MS to bacterial secretomes as demonstrated herein establishes proof-of-principle for use in novel microbial bioactive peptide discovery.",

author = "Kirkpatrick, \{Christine L.\} and Parsley, \{Nicole C.\} and Bartges, \{Tessa E.\} and Wing, \{Casey E.\} and Sushma Kommineni and Kristich, \{Christopher J.\} and Salzman, \{Nita H.\} and Patrie, \{Steven M.\} and Hicks, \{Leslie M.\}",

note = "Publisher Copyright: {\textcopyright} 2018 The Authors. Microbial Biotechnology published by John Wiley \& Sons Ltd and Society for Applied Microbiology.",

year = "2018",

month = sep,

doi = "10.1111/1751-7915.13299",

language = "English (US)",

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Kirkpatrick, CL, Parsley, NC, Bartges, TE, Wing, CE, Kommineni, S, Kristich, CJ, Salzman, NH, Patrie, SM & Hicks, LM 2018, 'Exploring bioactive peptides from bacterial secretomes using PepSAVI-MS: identification and characterization of Bac-21 from Enterococcus faecalis pPD1', Microbial Biotechnology, vol. 11, no. 5, pp. 943-951. https://doi.org/10.1111/1751-7915.13299

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AU - Parsley, Nicole C.

AU - Bartges, Tessa E.

AU - Wing, Casey E.

AU - Kommineni, Sushma

AU - Kristich, Christopher J.

AU - Salzman, Nita H.

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AU - Hicks, Leslie M.

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Exploring bioactive peptides from bacterial secretomes using PepSAVI-MS: identification and characterization of Bac-21 from Enterococcus faecalis pPD1

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