Escherichia coli mar and acrAB Mutants Display No Tolerance to Simple Alcohols

Ankarloo, Jonas; Wikman, Susanne; Nicholls, Ian A.

doi:10.3390/ijms11041403

Cited by 29 publications

(21 citation statements)

References 43 publications

(32 reference statements)

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“…The efflux pump protein mutants did not show any significant changes in growth inhibition compared to the wild type. This is consistent with literature reports which demonstrate that efflux pumps do not confer alcohol tolerance in E. coli [47,48]. The desaturase mutants, 7002ΔdesE and 7002ΔdesF, also do not deviate from the growth inhibition observed for the wild type; however, the desB mutant shows increased growth inhibition at 0.5 M ethanol (Figure 7.2B).…”

Section: Identifying Genes Responsible For Biofuel Tolerancesupporting

confidence: 81%

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Genetic engineering of cyanobacteria as biodiesel feedstock.

Ruffing¹,

Trahan²,

Jones³

2013

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Algal biofuels are a renewable energy source with the potential to replace conventional petroleum-based fuels, while simultaneously reducing greenhouse gas emissions. The economic feasibility of commercial algal fuel production, however, is limited by low productivity of the natural algal strains. The project described in this SAND report addresses this low algal productivity by genetically engineering cyanobacteria (i.e. blue-green algae) to produce free fatty acids as fuel precursors. The engineered strains were characterized using Sandia's unique imaging capabilities along with cutting-edge RNA-seq technology. These tools are applied to identify additional genetic targets for improving fuel production in cyanobacteria. This proofof-concept study demonstrates successful fuel production from engineered cyanobacteria, identifies potential limitations, and investigates several strategies to overcome these limitations. This project was funded from FY10-FY13 through the President Harry S. Truman Fellowship in National Security Science and Engineering, a program sponsored by the LDRD office at Sandia National Laboratories. 4 AcknowledgmentsThe authors of this SAND report acknowledge the support of management, particularly Anthony Martino, sponsor of Anne Ruffing's Truman Fellowship application; Eric Ackerman, Truman Fellow mentor; James Carney, Project Manager; Blake Simmons; and Ben Wu. We thank Michelle Raymer and Omar Garcia for their assistance with MCR analysis and operation of the hyperspectral confocal fluorescence microscope. We also acknowledge

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Section: Identifying Genes Responsible For Biofuel Tolerancesupporting

confidence: 81%

“…PCC7002: SYNPCC7002_A1013, SYNPCC7002_A0585, and SYNPCC7002_A0719. The efflux pump protein A1013 is homolgous to the AcrB efflux pump, which is responsible for solvent tolerance in E. coli [47]. The other two efflux pump proteins A0585 and A0719 are unique to Synechococcus sp.…”

Section: Identifying Genes Responsible For Biofuel Tolerancementioning

confidence: 99%

Genetic engineering of cyanobacteria as biodiesel feedstock.

Ruffing¹,

Trahan²,

Jones³

2013

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“…In the case of 1-octanol, deletion of acrA led to a threefold reduction in total levels of the final product attributed specifically to reduced levels in the media fraction, indicating that AcrAB-TolC exports this compound and is required for optimal production [77]. However, despite its broad substrate range this RND pump may be specific to hydrophobic compounds, as it does not provide tolerance toward more polar compounds such as short-chain alcohols [65,78]. Consistent with this, two studies report that deletion or inactivation of AcrB results in higher tolerance toward isobutanol [17,18].…”

Section: Reviewmentioning

confidence: 95%

Tolerance engineering in bacteria for the production of advanced biofuels and chemicals

Mukhopadhyay

2015

Trends in Microbiology

197

112

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During microbial production of solvent-like compounds, such as advanced biofuels and bulk chemicals, accumulation of the final product can negatively impact the cultivation of the host microbe and limit the production levels. Consequently, improving solvent tolerance is becoming an essential aspect of engineering microbial production strains. Mechanisms ranging from chaperones to transcriptional factors have been used to obtain solvent-tolerant strains. However, alleviating growth inhibition does not invariably result in increased production. Transporters specifically have emerged as a powerful category of proteins that bestow tolerance and often improve production but are difficult targets for cellular expression. Here we review strain engineering, primarily as it pertains to bacterial solvent tolerance, and the benefits and challenges associated with the expression of membrane-localized transporters in improving solvent tolerance and production.

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“…Similarly, but although not as well characterized, E. coli has been suggested to utilize the MdfA efflux pump to maintain its intracellular pH during fermentation (Krulwich et al 2005), although at this time there has been no work to indicate that any known efflux pump can confer increased tolerance to ethanol itself (Ankarloo et al 2010).…”

Section: Direct Chemical Effluxmentioning

confidence: 99%

Toxicological challenges to microbial bioethanol production and strategies for improved tolerance

et al. 2015

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Bioethanol production output has increased steadily over the last two decades and is now beginning to become competitive with traditional liquid transportation fuels due to advances in engineering, the identification of new production host organisms, and the development of novel biodesign strategies. A significant portion of these efforts has been dedicated to mitigating the toxicological challenges encountered across the bioethanol production process. From the release of potentially cytotoxic or inhibitory compounds from input feedstocks, through the metabolic co-synthesis of ethanol and potentially detrimental byproducts, and to the potential cytotoxicity of ethanol itself, each stage of bioethanol production requires the application of genetic or engineering controls that ensure the host organisms remain healthy and productive to meet the necessary economies required for large scale production. In addition, as production levels continue to increase, there is an escalating focus on the detoxification of the resulting waste streams to minimize their environmental impact. This review will present the major toxicological challenges encountered throughout each stage of the bioethanol production process and the commonly employed strategies for reducing or eliminating potential toxic effects.

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Escherichia coli mar and acrAB Mutants Display No Tolerance to Simple Alcohols

Cited by 29 publications

References 43 publications

Genetic engineering of cyanobacteria as biodiesel feedstock.

Genetic engineering of cyanobacteria as biodiesel feedstock.

Tolerance engineering in bacteria for the production of advanced biofuels and chemicals

Toxicological challenges to microbial bioethanol production and strategies for improved tolerance

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