Improving Mobilization of Foreign DNA into Zymomonas mobilis Strain ZM4 by Removal of Multiple Restriction Systems
Abstract
Zymomonas mobilis has emerged as a promising candidate for production of high-value bioproducts from plant biomass. However, a major limitation in equipping Z. mobilis with novel pathways to achieve this goal is restriction of heterologous DNA. Here, we characterized the contribution of several defense systems of Z. mobilis strain ZM4 to impeding heterologous gene transfer from an Escherichia coli donor. Bioinformatic analysis revealed that Z. mobilis ZM4 encodes a previously described mrr-like type IV restriction modification (RM) system, a type I-F CRISPR system, a chromosomal type I RM system (hsdMSc), and a previously uncharacterized type I RM system, located on an endogenous plasmid (hsdRMSp). The DNA recognition motif of HsdRMSp was identified by comparing the methylated DNA sequence pattern of mutants lacking one or both of the hsdMSc and hsdRMSp systems to that of the parent strain. The conjugation efficiency of synthetic plasmids containing single or combinations of the HsdMSc and HsdRMSp recognition sites indicated that both systems are active and decrease uptake of foreign DNA. In contrast, deletions of mrr and cas3 led to no detectable improvement in conjugation efficiency for the exogenous DNA tested. Thus, the suite of markerless restriction-negative strains that we constructed and the knowledge ofmore »
- Authors:
-
[1];
[2];
[4];
[1];
- DOE Great Lakes Bioenergy Research Center, University of Wisconsin—Madison, Madison, Wisconsin, USA, Department of Biomolecular Chemistry, University of Wisconsin—Madison, Madison, Wisconsin, USA
- DOE Great Lakes Bioenergy Research Center, University of Wisconsin—Madison, Madison, Wisconsin, USA, Wisconsin Energy Institute, University of Wisconsin—Madison, Madison, Wisconsin, USA
- Department of Biomolecular Chemistry, University of Wisconsin—Madison, Madison, Wisconsin, USA
- Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA, Center for Bioenergy Innovation, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA
- Publication Date:
- Research Org.:
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1819468
- Alternate Identifier(s):
- OSTI ID: 1824970
- Grant/Contract Number:
- FC-07ER64494; AC05-00OR22725
- Resource Type:
- Published Article
- Journal Name:
- Applied and Environmental Microbiology
- Additional Journal Information:
- Journal Name: Applied and Environmental Microbiology Journal Volume: 87 Journal Issue: 19; Journal ID: ISSN 0099-2240
- Publisher:
- American Society for Microbiology
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Zymomonas mobilis ZM4; restriction modification system; genome defense; conjugation efficiency of foreign genes; type I restriction enzymes
Citation Formats
Lal, Piyush Behari, Wells, Fritz, Myers, Kevin S., Banerjee, Rajdeep, Guss, Adam M., Kiley, Patricia J., and Kelly, ed., Robert M. Improving Mobilization of Foreign DNA into Zymomonas mobilis Strain ZM4 by Removal of Multiple Restriction Systems. United States: N. p., 2021.
Web. doi:10.1128/AEM.00808-21.
Lal, Piyush Behari, Wells, Fritz, Myers, Kevin S., Banerjee, Rajdeep, Guss, Adam M., Kiley, Patricia J., & Kelly, ed., Robert M. Improving Mobilization of Foreign DNA into Zymomonas mobilis Strain ZM4 by Removal of Multiple Restriction Systems. United States. https://doi.org/10.1128/AEM.00808-21
Lal, Piyush Behari, Wells, Fritz, Myers, Kevin S., Banerjee, Rajdeep, Guss, Adam M., Kiley, Patricia J., and Kelly, ed., Robert M. Fri .
"Improving Mobilization of Foreign DNA into Zymomonas mobilis Strain ZM4 by Removal of Multiple Restriction Systems". United States. https://doi.org/10.1128/AEM.00808-21.
@article{osti_1819468,
title = {Improving Mobilization of Foreign DNA into Zymomonas mobilis Strain ZM4 by Removal of Multiple Restriction Systems},
author = {Lal, Piyush Behari and Wells, Fritz and Myers, Kevin S. and Banerjee, Rajdeep and Guss, Adam M. and Kiley, Patricia J. and Kelly, ed., Robert M.},
abstractNote = {Zymomonas mobilis has emerged as a promising candidate for production of high-value bioproducts from plant biomass. However, a major limitation in equipping Z. mobilis with novel pathways to achieve this goal is restriction of heterologous DNA. Here, we characterized the contribution of several defense systems of Z. mobilis strain ZM4 to impeding heterologous gene transfer from an Escherichia coli donor. Bioinformatic analysis revealed that Z. mobilis ZM4 encodes a previously described mrr-like type IV restriction modification (RM) system, a type I-F CRISPR system, a chromosomal type I RM system (hsdMSc), and a previously uncharacterized type I RM system, located on an endogenous plasmid (hsdRMSp). The DNA recognition motif of HsdRMSp was identified by comparing the methylated DNA sequence pattern of mutants lacking one or both of the hsdMSc and hsdRMSp systems to that of the parent strain. The conjugation efficiency of synthetic plasmids containing single or combinations of the HsdMSc and HsdRMSp recognition sites indicated that both systems are active and decrease uptake of foreign DNA. In contrast, deletions of mrr and cas3 led to no detectable improvement in conjugation efficiency for the exogenous DNA tested. Thus, the suite of markerless restriction-negative strains that we constructed and the knowledge of this new restriction system and its DNA recognition motif provide the necessary platform to flexibly engineer the next generation of Z. mobilis strains for synthesis of valuable products.},
doi = {10.1128/AEM.00808-21},
journal = {Applied and Environmental Microbiology},
number = 19,
volume = 87,
place = {United States},
year = {Fri Sep 10 00:00:00 EDT 2021},
month = {Fri Sep 10 00:00:00 EDT 2021}
}
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