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Title: Acute Limonene Toxicity in Escherichia coli Is Caused by Limonene Hydroperoxide and Alleviated by a Point Mutation in Alkyl Hydroperoxidase AhpC

Abstract

ABSTRACT Limonene, a prime component of citrus peel oil, has a number of applications related to microbiology. The antimicrobial properties of limonene make it a popular disinfectant and food preservative, while its possibility as a biofuel component has made it the target of renewable production efforts through microbial metabolic engineering. For both applications, an understanding of microbial sensitivity or tolerance to limonene is crucial, but the mechanism of limonene toxicity remains enigmatic. In this study, we characterized a limonene-tolerant strain ofEscherichia coliand found a mutation inahpC, encoding alkyl hydroperoxidase, which alleviated limonene toxicity. We demonstrate that the acute toxicity previously attributed to limonene is largely due to the common oxidation product limonene hydroperoxide, which forms spontaneously in aerobic environments. The mutant AhpC protein with an L-to-Q change at position 177 (AhpC L177Q) was able to alleviate this toxicity by reducing the hydroperoxide to a more benign compound. We reveal that the degree of limonene toxicity is a function of its oxidation level and that nonoxidized limonene has relatively little toxicity to wild-typeE. colicells. Our results have implications for both the renewable production of limonene and the applications of limonene as an antimicrobial.

Authors:
 [1];  [2];  [3];  [1];  [1];  [4];  [1];  [4];  [1]
  1. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  2. Total New Energies USA, Inc., Emeryville, CA (United States)
  3. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
  4. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1512098
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Applied and Environmental Microbiology
Additional Journal Information:
Journal Volume: 81; Journal Issue: 14; Journal ID: ISSN 0099-2240
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Chubukov, Victor, Mingardon, Florence, Schackwitz, Wendy, Baidoo, Edward E. K., Alonso-Gutierrez, Jorge, Hu, Qijun, Lee, Taek Soon, Keasling, Jay D., and Mukhopadhyay, Aindrila. Acute Limonene Toxicity in Escherichia coli Is Caused by Limonene Hydroperoxide and Alleviated by a Point Mutation in Alkyl Hydroperoxidase AhpC. United States: N. p., 2015. Web. doi:10.1128/AEM.01102-15.
Chubukov, Victor, Mingardon, Florence, Schackwitz, Wendy, Baidoo, Edward E. K., Alonso-Gutierrez, Jorge, Hu, Qijun, Lee, Taek Soon, Keasling, Jay D., & Mukhopadhyay, Aindrila. Acute Limonene Toxicity in Escherichia coli Is Caused by Limonene Hydroperoxide and Alleviated by a Point Mutation in Alkyl Hydroperoxidase AhpC. United States. doi:10.1128/AEM.01102-15.
Chubukov, Victor, Mingardon, Florence, Schackwitz, Wendy, Baidoo, Edward E. K., Alonso-Gutierrez, Jorge, Hu, Qijun, Lee, Taek Soon, Keasling, Jay D., and Mukhopadhyay, Aindrila. Fri . "Acute Limonene Toxicity in Escherichia coli Is Caused by Limonene Hydroperoxide and Alleviated by a Point Mutation in Alkyl Hydroperoxidase AhpC". United States. doi:10.1128/AEM.01102-15. https://www.osti.gov/servlets/purl/1512098.
@article{osti_1512098,
title = {Acute Limonene Toxicity in Escherichia coli Is Caused by Limonene Hydroperoxide and Alleviated by a Point Mutation in Alkyl Hydroperoxidase AhpC},
author = {Chubukov, Victor and Mingardon, Florence and Schackwitz, Wendy and Baidoo, Edward E. K. and Alonso-Gutierrez, Jorge and Hu, Qijun and Lee, Taek Soon and Keasling, Jay D. and Mukhopadhyay, Aindrila},
abstractNote = {ABSTRACT Limonene, a prime component of citrus peel oil, has a number of applications related to microbiology. The antimicrobial properties of limonene make it a popular disinfectant and food preservative, while its possibility as a biofuel component has made it the target of renewable production efforts through microbial metabolic engineering. For both applications, an understanding of microbial sensitivity or tolerance to limonene is crucial, but the mechanism of limonene toxicity remains enigmatic. In this study, we characterized a limonene-tolerant strain ofEscherichia coliand found a mutation inahpC, encoding alkyl hydroperoxidase, which alleviated limonene toxicity. We demonstrate that the acute toxicity previously attributed to limonene is largely due to the common oxidation product limonene hydroperoxide, which forms spontaneously in aerobic environments. The mutant AhpC protein with an L-to-Q change at position 177 (AhpCL177Q) was able to alleviate this toxicity by reducing the hydroperoxide to a more benign compound. We reveal that the degree of limonene toxicity is a function of its oxidation level and that nonoxidized limonene has relatively little toxicity to wild-typeE. colicells. Our results have implications for both the renewable production of limonene and the applications of limonene as an antimicrobial.},
doi = {10.1128/AEM.01102-15},
journal = {Applied and Environmental Microbiology},
number = 14,
volume = 81,
place = {United States},
year = {2015},
month = {5}
}

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