The change of the state of cell membrane can enhance the synthesis of menaquinone in Escherichia coli

Liu, Yan; Ding, Xiu-min; Xue, Zhenglian; Hu, Liu-xiu; Zhang, Ning-juan; Zhou, Wang; Yang, Jianwei; Cheng, Qian; Chen, Minghong; Zhang, Zhuang-zhuang; Zheng, Zhiming

doi:10.1007/s11274-017-2222-9

Cited by 13 publications

(10 citation statements)

References 27 publications

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“…Maintenance of appropriate membrane fluidity is vital to membrane function. In general, membrane fluidity trends with the relative abundance of unsaturated lipids (43), though other membrane components can also contribute to this property (16,44,45). Membrane fluidity was measured via cell membrane fluorescence polarization, where increased polarization corresponds to decreased membrane fluidity (9,46).…”

Section: Resultsmentioning

confidence: 99%

Lessons in Membrane Engineering for Octanoic Acid Production from Environmental Escherichia coli Isolates

Chen

Reinhardt

Neris

et al. 2018

Appl Environ Microbiol

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Fermentative production of many attractive biorenewable fuels and chemicals is limited by product toxicity in the form of damage to the microbial cell membrane. Metabolic engineering of the production organism can help mitigate this problem, but there is a need for identification and prioritization of the most effective engineering targets. Here, we use a set of previously characterized environmental isolates with high tolerance and production of octanoic acid, a model membrane-damaging biorenewable product, as a case study for identifying and prioritizing membrane engineering strategies. This characterization identified differences in the membrane lipid composition, fluidity, integrity, and cell surface hydrophobicity from those of the lab strain MG1655. Consistent with previous publications, decreased membrane fluidity was associated with increased fatty acid production ability. Maintenance of high membrane integrity or longer membrane lipids seemed to be of less importance than fluidity. Cell surface hydrophobicity was also directly associated with fatty acid production titers, with the strength of this association demonstrated by plasmid-based expression of the multiple stress resistance outer membrane protein BhsA. This expression of was effective in altering hydrophobicity, but the direction and magnitude of the change differed between strains. Thus, additional strategies are needed to reliably engineer cell surface hydrophobicity. This work demonstrates the ability of environmental microbiological studies to impact the metabolic engineering design-build-test-learn cycle and possibly increase the economic viability of fermentative bioprocesses. The production of bulk fuels and chemicals in a bio-based fermentation process requires high product titers. This is often difficult to achieve, because many of the target molecules damage the membrane of the microbial cell factory. Engineering the composition of the membrane in order to decrease its vulnerability to this damage has proven to be an effective strategy for improving bioproduction, but additional strategies and engineering targets are needed. Here, we studied a small set of environmental isolates that have higher production titers of octanoic acid, a model biorenewable chemical, than those of the lab strain MG1655. We found that membrane fluidity and cell surface hydrophobicity are strongly associated with improved octanoic acid production. Fewer genetic modification strategies have been demonstrated for tuning hydrophobicity relative to fluidity, leading to the conclusion that there is a need for expanding hydrophobicity engineering strategies in.

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Section: Resultsmentioning

confidence: 99%

Lessons in Membrane Engineering for Octanoic Acid Production from Environmental Escherichia coli Isolates

Chen

Reinhardt

Neris

et al. 2018

Appl Environ Microbiol

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“…It was, however, apparent that the increase in MK–7 concentration is not merely a reflection on cell density. Binding of IONs@APTES to B. subtilis might have changed the state or composition of cell membranes, resulting in enhanced secretion of MK–7 to the fermentation medium [ 7 , 20 ].…”

Section: Resultsmentioning

confidence: 99%

Impact of 3–Aminopropyltriethoxysilane-Coated Iron Oxide Nanoparticles on Menaquinone-7 Production Using B. subtilis

et al. 2017

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One of the major issues associated with industrial production of menaquinone-7 (MK–7) is the low fermentation yield. In this study, we investigated the effect of iron oxide nanoparticles coated with 3–aminopropyltriethoxysilane (IONs@APTES) on the production of MK–7 using B. subtilis (ATCC 6633). Decoration of B. subtilis cells with IONs@APTES significantly enhanced both MK–7 production and yield. An approximately two-fold increase in MK–7 production (41 mg/L) was observed in the presence of 500 µg/mL IONs@APTES, as compared to MK–7 production using untreated bacteria (22 mg/L). This paper, therefore, illustrates the immense biotechnological potential of IONs@APTES in increasing MK–7 concentration using B. subtilis, and its future role in bioprocess engineering.

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“…Transcriptomic analysis showed that the pathways such as ABC transporters, bacterial chemotaxis, and oxidative phosphorylation changed significantly, indicating that the respiratory system and state of the cell membrane were changed obviously in SinR quad . A previous study discovered that changing the state of the membrane in Escherichia coli could increase the synthesis of MK-8 [ 62 ]. Our study verified this prediction by quantitatively determining the membrane potential and respiration of SinR quad and BS168.…”

Section: Discussionmentioning

confidence: 99%

Site-directed mutagenesis of the quorum-sensing transcriptional regulator SinR affects the biosynthesis of menaquinone in Bacillus subtilis

Zhao

et al. 2021

Microb Cell Fact

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Background Menaquinone (MK-7) is a highly valuable vitamin K2 produced by Bacillus subtilis. Common static metabolic engineering approaches for promoting the production of MK-7 have been studied previously. However, these approaches caused an accumulation of toxic substances and reduced product yield. Hence, dynamic regulation by the quorum sensing (QS) system is a promising method for achieving a balance between product synthesis and cell growth. Results In this study, the QS transcriptional regulator SinR, which plays a significant role in biofilm formation and MK production simultaneously, was selected, and its site-directed mutants were constructed. Among these mutants, sinR knock out strain (KO-SinR) increased the biofilm biomass by 2.8-fold compared to the wild-type. SinRquad maximized the yield of MK-7 (102.56 ± 2.84 mg/L). To decipher the mechanism of how this mutant regulates MK-7 synthesis and to find additional potential regulators that enhance MK-7 synthesis, RNA-seq was used to analyze expression changes in the QS system, biofilm formation, and MK-7 synthesis pathway. The results showed that the expressions of tapA, tasA and epsE were up-regulated 9.79-, 0.95-, and 4.42-fold, respectively. Therefore, SinRquad formed more wrinkly and smoother biofilms than BS168. The upregulated expressions of glpF, glpk, and glpD in this biofilm morphology facilitated the flow of glycerol through the biofilm. In addition, NADH dehydrogenases especially sdhA, sdhB, sdhC and glpD, increased 1.01-, 3.93-, 1.87-, and 1.11-fold, respectively. The increased expression levels of NADH dehydrogenases indicated that more electrons were produced for the electron transport system. Electrical hyperpolarization stimulated the synthesis of the electron transport chain components, such as cytochrome c and MK, to ensure the efficiency of electron transfer. Wrinkly and smooth biofilms formed a network of interconnected channels with a low resistance to liquid flow, which was beneficial for the uptake of glycerol, and facilitated the metabolic flux of four modules of the MK-7 synthesis pathway. Conclusions In this study, we report for the first time that SinRquad has significant effects on MK-7 synthesis by forming wrinkly and smooth biofilms, upregulating the expression level of most NADH dehydrogenases, and providing higher membrane potential to stimulate the accumulation of the components in the electron transport system.

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The change of the state of cell membrane can enhance the synthesis of menaquinone in Escherichia coli

Cited by 13 publications

References 27 publications

Lessons in Membrane Engineering for Octanoic Acid Production from Environmental Escherichia coli Isolates

Lessons in Membrane Engineering for Octanoic Acid Production from Environmental Escherichia coli Isolates

Impact of 3–Aminopropyltriethoxysilane-Coated Iron Oxide Nanoparticles on Menaquinone-7 Production Using B. subtilis

Site-directed mutagenesis of the quorum-sensing transcriptional regulator SinR affects the biosynthesis of menaquinone in Bacillus subtilis

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