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Title: Enhanced fatty acid production in engineered chemolithoautotrophic bacteria using reduced sulfur compounds as energy sources

Abstract

Chemolithoautotrophic bacteria that oxidize reduced sulfur compounds, such as H 2 S, while fixing CO 2 are an untapped source of renewable bioproducts from sulfide-laden waste, such as municipal wastewater. In this study, we report engineering of the chemolithoautotrophic bacterium Thiobacillus denitrificans to produce up to 52-fold more fatty acids than the wild-type strain when grown with thiosulfate and CO 2 . A modified thioesterase gene from E. coli ('tesA) was integrated into the T. denitrificans chromosome under the control of P kan or one of two native T. denitrificans promoters. The relative strength of the two native promoters as assessed by fatty acid production in engineered strains was very similar to that assessed by expression of the cognate genes in the wild-type strain. This proof-of-principle study suggests that engineering sulfide-oxidizing chemolithoautotrophic bacteria to overproduce fatty acid-derived products merits consideration as a technology that could simultaneously produce renewable fuels/chemicals as well as cost-effectively remediate sulfide-contaminated wastewater.

Authors:
 [1];  [2];  [2];  [3];  [4]
  1. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Earth and Environmental Sciences
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Earth and Environmental Sciences
  3. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering
  4. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering; Univ. of California, Berkeley, CA (United States). Dept. of Chemical and Biomolecular Engineering and of Bioengineering; Technical Univ. of Denmark, Horsholm (Denmark). Novo Nordisk Foundation Center for Biosustainability
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:
1394664
Alternate Identifier(s):
OSTI ID: 1393092
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article: Published Article
Journal Name:
Metabolic Engineering Communications
Additional Journal Information:
Journal Volume: 3; Journal Issue: C; Journal ID: ISSN 2214-0301
Publisher:
Elsevier
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; chemolithoautotrophic; sulfide; fatty acids; tesA; thiobacillus denitrificans

Citation Formats

Beller, Harry R., Zhou, Peng, Jewell, Talia N. M., Goh, Ee-Been, and Keasling, Jay D.. Enhanced fatty acid production in engineered chemolithoautotrophic bacteria using reduced sulfur compounds as energy sources. United States: N. p., 2016. Web. doi:10.1016/j.meteno.2016.07.001.
Beller, Harry R., Zhou, Peng, Jewell, Talia N. M., Goh, Ee-Been, & Keasling, Jay D.. Enhanced fatty acid production in engineered chemolithoautotrophic bacteria using reduced sulfur compounds as energy sources. United States. doi:10.1016/j.meteno.2016.07.001.
Beller, Harry R., Zhou, Peng, Jewell, Talia N. M., Goh, Ee-Been, and Keasling, Jay D.. Tue . "Enhanced fatty acid production in engineered chemolithoautotrophic bacteria using reduced sulfur compounds as energy sources". United States. doi:10.1016/j.meteno.2016.07.001.
@article{osti_1394664,
title = {Enhanced fatty acid production in engineered chemolithoautotrophic bacteria using reduced sulfur compounds as energy sources},
author = {Beller, Harry R. and Zhou, Peng and Jewell, Talia N. M. and Goh, Ee-Been and Keasling, Jay D.},
abstractNote = {Chemolithoautotrophic bacteria that oxidize reduced sulfur compounds, such as H 2 S, while fixing CO 2 are an untapped source of renewable bioproducts from sulfide-laden waste, such as municipal wastewater. In this study, we report engineering of the chemolithoautotrophic bacterium Thiobacillus denitrificans to produce up to 52-fold more fatty acids than the wild-type strain when grown with thiosulfate and CO 2 . A modified thioesterase gene from E. coli ('tesA) was integrated into the T. denitrificans chromosome under the control of P kan or one of two native T. denitrificans promoters. The relative strength of the two native promoters as assessed by fatty acid production in engineered strains was very similar to that assessed by expression of the cognate genes in the wild-type strain. This proof-of-principle study suggests that engineering sulfide-oxidizing chemolithoautotrophic bacteria to overproduce fatty acid-derived products merits consideration as a technology that could simultaneously produce renewable fuels/chemicals as well as cost-effectively remediate sulfide-contaminated wastewater.},
doi = {10.1016/j.meteno.2016.07.001},
journal = {Metabolic Engineering Communications},
number = C,
volume = 3,
place = {United States},
year = {Tue Jul 05 00:00:00 EDT 2016},
month = {Tue Jul 05 00:00:00 EDT 2016}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.1016/j.meteno.2016.07.001

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