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Title: Development of Chemical and Metabolite Sensors for Rhodococcus opacus PD630

Rhodococcus opacus PD630 is a non-model, gram positive bacterium that possesses desirable traits for biomass conversion, including consumption capabilities for lignocellulose-based sugars and toxic lignin-derived aromatic compounds, significant triacylglycerol accumulation, relatively rapid growth rate, and genetic tractability. However, few genetic elements have been directly characterized in R. opacus, limiting its application for lignocellulose bioconversion. Here, we report the characterization and development of genetic tools for tunable gene expression in R. opacus, including: 1) six fluorescent reporters for quantifying promoter output, 2) three chemically inducible promoters for variable gene expression, and 3) two classes of metabolite sensors derived from native R. opacus promoters that detect nitrogen levels or aromatic compounds. Using these tools, we also provide insights into native aromatic consumption pathways in R. opacus. Overall, this work expands the ability to control and characterize gene expression in R. opacus for future lignocellulose-based fuel and chemical production.
Authors:
 [1] ;  [1] ; ORCiD logo [1]
  1. Washington Univ., St. Louis, MO (United States)
Publication Date:
Grant/Contract Number:
SC0012705
Type:
Accepted Manuscript
Journal Name:
ACS Synthetic Biology
Additional Journal Information:
Journal Volume: 6; Journal Issue: 10; Journal ID: ISSN 2161-5063
Publisher:
American Chemical Society (ACS)
Research Org:
Washington Univ., St. Louis, MO (United States)
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; Rhodococcus opacus; lignin; promoter; sensor; nitrogen; aromatic
OSTI Identifier:
1417982

DeLorenzo, Drew M., Henson, William R., and Moon, Tae Seok. Development of Chemical and Metabolite Sensors for Rhodococcus opacus PD630. United States: N. p., Web. doi:10.1021/acssynbio.7b00192.
DeLorenzo, Drew M., Henson, William R., & Moon, Tae Seok. Development of Chemical and Metabolite Sensors for Rhodococcus opacus PD630. United States. doi:10.1021/acssynbio.7b00192.
DeLorenzo, Drew M., Henson, William R., and Moon, Tae Seok. 2017. "Development of Chemical and Metabolite Sensors for Rhodococcus opacus PD630". United States. doi:10.1021/acssynbio.7b00192. https://www.osti.gov/servlets/purl/1417982.
@article{osti_1417982,
title = {Development of Chemical and Metabolite Sensors for Rhodococcus opacus PD630},
author = {DeLorenzo, Drew M. and Henson, William R. and Moon, Tae Seok},
abstractNote = {Rhodococcus opacus PD630 is a non-model, gram positive bacterium that possesses desirable traits for biomass conversion, including consumption capabilities for lignocellulose-based sugars and toxic lignin-derived aromatic compounds, significant triacylglycerol accumulation, relatively rapid growth rate, and genetic tractability. However, few genetic elements have been directly characterized in R. opacus, limiting its application for lignocellulose bioconversion. Here, we report the characterization and development of genetic tools for tunable gene expression in R. opacus, including: 1) six fluorescent reporters for quantifying promoter output, 2) three chemically inducible promoters for variable gene expression, and 3) two classes of metabolite sensors derived from native R. opacus promoters that detect nitrogen levels or aromatic compounds. Using these tools, we also provide insights into native aromatic consumption pathways in R. opacus. Overall, this work expands the ability to control and characterize gene expression in R. opacus for future lignocellulose-based fuel and chemical production.},
doi = {10.1021/acssynbio.7b00192},
journal = {ACS Synthetic Biology},
number = 10,
volume = 6,
place = {United States},
year = {2017},
month = {7}
}