Rapid Screening of Lanthipeptide Analogs via In-Colony Removal of Leader Peptides in Escherichia coli

Si, Tong; Tian, Qiqi; Min, Yuhao; Zhang, Linzixuan; Sweedler, Jonathan V.; van der Donk, Wilfred A.; Zhao, Huimin

doi:10.1021/jacs.8b05544

Title: Rapid Screening of Lanthipeptide Analogs via In-Colony Removal of Leader Peptides in Escherichia coli

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Abstract

Most native producers of ribosomally synthesized and post-translationally modified peptides (RiPPs) utilize N-terminal leader peptides to avoid potential cytotoxicity of mature products to the hosts. Unfortunately, the native machinery of leader peptide removal is often difficult to reconstitute in heterologous hosts. Here in this paper we devised a general method to produce bioactive lanthipeptides, a major class of RiPP molecules, in Escherichia coli colonies using synthetic biology principles, where leader peptide removal is programmed temporally by protease compartmentalization and inducible cell autolysis. We demonstrated the method for producing two lantibiotics, haloduracin and lacticin 481, and performed analog screening for haloduracin. This method enables facile, high throughput discovery, characterization, and engineering of RiPPs.

Authors:

^[1]; Tian, Qiqi ^[2]; Min, Yuhao ^[3]; Zhang, Linzixuan ^[3];

^[4];

^[5];

^[6]

Univ. of Illinois, Urbana-Champaign, IL (United States). Carl R. Woese Inst. for Genomic Biology
Univ. of Illinois, Urbana-Champaign, IL (United States). Dept. of Biochemistry
Univ. of Illinois, Urbana-Champaign, IL (United States). Dept. of Chemistry
Univ. of Illinois, Urbana-Champaign, IL (United States). Carl R. Woese Inst. for Genomic Biology, and Dept. of Chemistry, and Beckman Inst. for Advanced Science and Technology
Univ. of Illinois, Urbana-Champaign, IL (United States). Carl R. Woese Inst. for Genomic Biology, and Dept. of Chemistry, and Howard Hughes Medical Inst.
Univ. of Illinois, Urbana-Champaign, IL (United States). Carl R. Woese Inst. for Genomic Biology, and Dept. of Biochemistry, Dept. of Chemistry, Dept. of Chemical and Biomolecular Engineering, and Dept. of Bioengineering

Publication Date:: Wed Sep 12 00:00:00 EDT 2018

Research Org.:: Univ. of Illinois, Champaign, IL (United States). Center for Advanced Bioenergy and Bioproducts Innovation (CABBI); Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)

Sponsoring Org.:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

OSTI Identifier:: 1471403

Alternate Identifier(s):: OSTI ID: 1991824

Grant/Contract Number:: SC0018420

Resource Type:: Accepted Manuscript

Journal Name:: Journal of the American Chemical Society

Additional Journal Information:: Journal Volume: 140; Journal Issue: 38; Journal ID: ISSN 0002-7863

Publisher:: American Chemical Society (ACS)

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES

Citation Formats


                    Si, Tong, Tian, Qiqi, Min, Yuhao, Zhang, Linzixuan, Sweedler, Jonathan V., van der Donk, Wilfred A., and Zhao, Huimin. Rapid Screening of Lanthipeptide Analogs via In-Colony Removal of Leader Peptides in Escherichia coli.  United States: N. p., 2018. 
Web.  doi:10.1021/jacs.8b05544.

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                    Si, Tong, Tian, Qiqi, Min, Yuhao, Zhang, Linzixuan, Sweedler, Jonathan V., van der Donk, Wilfred A., & Zhao, Huimin. Rapid Screening of Lanthipeptide Analogs via In-Colony Removal of Leader Peptides in Escherichia coli.  United States.  https://doi.org/10.1021/jacs.8b05544

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                    Si, Tong, Tian, Qiqi, Min, Yuhao, Zhang, Linzixuan, Sweedler, Jonathan V., van der Donk, Wilfred A., and Zhao, Huimin. Wed .  
"Rapid Screening of Lanthipeptide Analogs via In-Colony Removal of Leader Peptides in Escherichia coli".  United States.  https://doi.org/10.1021/jacs.8b05544.  https://www.osti.gov/servlets/purl/1471403.

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@article{osti_1471403,

  title        = {Rapid Screening of Lanthipeptide Analogs via In-Colony Removal of Leader Peptides in Escherichia coli},

  author       = {Si, Tong and Tian, Qiqi and Min, Yuhao and Zhang, Linzixuan and Sweedler, Jonathan V. and van der Donk, Wilfred A. and Zhao, Huimin},

  abstractNote = {Most native producers of ribosomally synthesized and post-translationally modified peptides (RiPPs) utilize N-terminal leader peptides to avoid potential cytotoxicity of mature products to the hosts. Unfortunately, the native machinery of leader peptide removal is often difficult to reconstitute in heterologous hosts. Here in this paper we devised a general method to produce bioactive lanthipeptides, a major class of RiPP molecules, in Escherichia coli colonies using synthetic biology principles, where leader peptide removal is programmed temporally by protease compartmentalization and inducible cell autolysis. We demonstrated the method for producing two lantibiotics, haloduracin and lacticin 481, and performed analog screening for haloduracin. This method enables facile, high throughput discovery, characterization, and engineering of RiPPs.},

  doi          = {10.1021/jacs.8b05544},

  journal      = {Journal of the American Chemical Society},

  number       = 38,

  volume       = 140,

  place        = {United States},

  year         = {Wed Sep 12 00:00:00 EDT 2018},

  month        = {Wed Sep 12 00:00:00 EDT 2018}

}

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