Engineering the Microbial Cell Membrane To Improve Bioproduction

Jarboe, Laura R.; Klauda, Jeffery B.; Chen, Yingxi; Davis, Kirsten; Santoscoy, Miguel C.

doi:10.1021/bk-2018-1310.ch003

Cited by 5 publications

(6 citation statements)

References 118 publications

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“…The applicability of this assay is further demonstrated through the characterization of four membrane engineering strategies that aim to increase tolerance to exogenous octanoic acid (C8). The damaging effects of carboxylic acids on the integrity of microbial cell membrane have been previously and extensively described [9,[14][15][16]53]. The in situ SYTOX assay described here allows monitoring of changes in the membrane permeability in real time without a priori selection of sampling times (Fig 4).…”

Section: In Situ Assessment Of Membrane Engineering Strategiesmentioning

confidence: 99%

“…One strategy for addressing this problem is to increase the robustness of the production organism to the problematic inhibitor, such as through evolutionary or targeted strain development, [3,[5][6][7]. In many cases, damage of the microbial cell membrane is a substantial component of the microbial inhibition, and thus the membrane is an attractive engineering target [8][9][10][11][12][13][14][15][16][17]. Engineering of the microbial cell membrane to combat this damage has been demonstrated, for example, to improve production of fatty acids [18][19][20][21], succinate [22], styrene [21] and hyaluronic acid [23].…”

Section: Introductionmentioning

confidence: 99%

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Streamlined assessment of membrane permeability and its application to membrane engineering of Escherichia coli for octanoic acid tolerance

Santoscoy

Jarboe

2019

Journal of Industrial Microbiology and Biotechnology

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The economic viability of bio-production processes is often limited by damage to the microbial cell membrane and thus there is a demand for strategies to increase the robustness of the cell membrane. Damage to the microbial membrane is also a common mode of action by antibiotics. Membrane-impermeable DNA-binding dyes are often used to assess membrane integrity in conjunction with flow cytometry. We demonstrate that in situ assessment of the membrane permeability of E. coli to SYTOX Green is consistent with flow cytometry, with the benefit of lower experimental intensity, lower cost, and no need for a priori selection of sampling times. This method is demonstrated by the characterization of four membrane engineering strategies (deletion of aas, deletion of cfa, increased expression of cfa, and deletion of bhsA) for their effect on octanoic acid tolerance, with the finding that deletion of bhsA increased tolerance and substantially decreased membrane leakage. ABSTRACTThe economic viability of bio-production processes is often limited by damage to the microbial cell membrane and thus there is a demand for strategies to increase the robustness of the cell membrane. Damage to the microbial membrane is also a common mode of action by antibiotics. Membrane-impermeable DNA-binding dyes are often used to assess membrane integrity in conjunction with flow cytometry. We demonstrate that in situ assessment of the membrane permeability of E. coli to SYTOX Green is consistent with flow cytometry, with the benefit of lower experimental intensity, lower cost, and no need for a priori selection of sampling times. This method is demonstrated by the characterization of four membrane engineering strategies (deletion of aas, deletion of cfa, increased expression of cfa, and deletion of bhsA) for their effect on octanoic acid tolerance, with the finding that deletion of bhsA increased tolerance and substantially decreased membrane leakage.

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Section: In Situ Assessment Of Membrane Engineering Strategiesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Streamlined assessment of membrane permeability and its application to membrane engineering of Escherichia coli for octanoic acid tolerance

Santoscoy

Jarboe

2019

Journal of Industrial Microbiology and Biotechnology

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“…Microbial attachment to surfaces is known to be impacted by variation in cell surface properties, such as hydrophobicity and the presence and abundance of various proteins and sugars [ 1 ]. Some of these same cell surface properties have also been implicated in microbial tolerance to harsh growth conditions [ 2 – 4 ]. It is desirable to be able to predictably engineer the properties of the microbial cell membrane for biotechnology applications [ 2 , 5 – 7 ].…”

Section: Introductionmentioning

confidence: 99%

Allelic variation of Escherichia coli outer membrane protein A: Impact on cell surface properties, stress tolerance and allele distribution

Liao

Santoscoy²,

Craft³

et al. 2022

PLoS ONE

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Outer membrane protein A (OmpA) is one of the most abundant outer membrane proteins of Gram-negative bacteria and is known to have patterns of sequence variations at certain amino acids—allelic variation—in Escherichia coli. Here we subjected seven exemplar OmpA alleles expressed in a K-12 (MG1655) ΔompA background to further characterization. These alleles were observed to significantly impact cell surface charge (zeta potential), cell surface hydrophobicity, biofilm formation, sensitivity to killing by neutrophil elastase, and specific growth rate at 42°C and in the presence of acetate, demonstrating that OmpA is an attractive target for engineering cell surface properties and industrial phenotypes. It was also observed that cell surface charge and biofilm formation both significantly correlate with cell surface hydrophobicity, a cell property that is increasingly intriguing for bioproduction. While there was poor alignment between the observed experimental values relative to the known sequence variation, differences in hydrophobicity and biofilm formation did correspond to the identity of residue 203 (N vs T), located within the proposed dimerization domain. The relative abundance of the (I, δ) allele was increased in extraintestinal pathogenic E. coli (ExPEC) isolates relative to environmental isolates, with a corresponding decrease in (I, α) alleles in ExPEC relative to environmental isolates. The (I, α) and (I, δ) alleles differ at positions 203 and 251. Variations in distribution were also observed among ExPEC types and phylotypes. Thus, OmpA allelic variation and its influence on OmpA function warrant further investigation.

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“…Manipulation of the cell membrane composition is a common approach to increase microbial solvent tolerance, and can be achieved through genetic engineering, 7 for example through increasing the proportion of saturated hydrocarbon tails of phospholipids to reduce membrane fluidity. 8 Microbial membrane fluidity can also be influenced by the presence of molecules that intercalate the membrane.…”

Section: Introductionmentioning

confidence: 99%

Carotenoids improve bacterial tolerance towards biobutanol through membrane stabilization

Chia

Seviour

Kjelleberg

et al. 2021

Environ. Sci.: Nano

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Engineering the Microbial Cell Membrane To Improve Bioproduction

Cited by 5 publications

References 118 publications

Streamlined assessment of membrane permeability and its application to membrane engineering of Escherichia coli for octanoic acid tolerance

Streamlined assessment of membrane permeability and its application to membrane engineering of Escherichia coli for octanoic acid tolerance

Allelic variation of Escherichia coli outer membrane protein A: Impact on cell surface properties, stress tolerance and allele distribution

Carotenoids improve bacterial tolerance towards biobutanol through membrane stabilization

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