Display of membrane proteins on the heterologous caveolae carved by caveolin-1 in the Escherichia coli cytoplasm

Shin, Ji Yeon; Yoon, Jinyoung; Cho, Da Hyeong; Park, Myungseo; Lee, Jun Young; Yang, Yoosoo; Jeong, Cherlhyun; Sung, Bong Hyun; Sohn, Jae Hak; Park, Jin Byung; Kweon, Dae Hyuk

doi:10.1016/j.enzmictec.2015.06.018

Cited by 17 publications

(10 citation statements)

References 31 publications

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“…Another related question addresses the co-overexpression of two membrane proteins, with one of them being able to induce neo-formed intracellular membranes, as reported for b and c subunits of the F 1 F 0 -ATP synthase [ 50 ], and which is proposed for other membrane proteins in association with the b subunit [ 113 ] or with Cav-1 [ 114 ]. The questions about the fate of the second membrane protein and the ways to predict it are intriguing.…”

Section: Some Remaining Questionsmentioning

confidence: 99%

Ectopic Neo-Formed Intracellular Membranes in Escherichia coli: A Response to Membrane Protein-Induced Stress Involving Membrane Curvature and Domains

et al. 2018

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Bacterial cytoplasmic membrane stress induced by the overexpression of membrane proteins at high levels can lead to formation of ectopic intracellular membranes. In this review, we report the various observations of such membranes in Escherichia coli, compare their morphological and biochemical characterizations, and we analyze the underlying molecular processes leading to their formation. Actually, these membranes display either vesicular or tubular structures, are separated or connected to the cytoplasmic membrane, present mono- or polydispersed sizes and shapes, and possess ordered or disordered arrangements. Moreover, their composition differs from that of the cytoplasmic membrane, with high amounts of the overexpressed membrane protein and altered lipid-to-protein ratio and cardiolipin content. These data reveal the importance of membrane domains, based on local specific lipid–protein and protein–protein interactions, with both being crucial for local membrane curvature generation, and they highlight the strong influence of protein structure. Indeed, whether the cylindrically or spherically curvature-active proteins are actively curvogenic or passively curvophilic, the underlying molecular scenarios are different and can be correlated with the morphological features of the neo-formed internal membranes. Delineating these molecular mechanisms is highly desirable for a better understanding of protein–lipid interactions within membrane domains, and for optimization of high-level membrane protein production in E. coli.

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Section: Some Remaining Questionsmentioning

confidence: 99%

Ectopic Neo-Formed Intracellular Membranes in Escherichia coli: A Response to Membrane Protein-Induced Stress Involving Membrane Curvature and Domains

et al. 2018

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“…[ 43 ] Heterologous expression of cav3 has not yet been reported. Cav1 has also been employed to increase the accumulation of membrane proteins in E. coli [ 21 ] or to enhance the uptake and conversion of fatty acids into esters. [ 22 ] We thus examined whether the production of rainbow colorants can be enhanced by expanding membrane structures by generation of IMVs through employing caveolae.…”

Section: Resultsmentioning

confidence: 99%

“…Several recent studies attempted to resolve this issue by membrane engineering [ 17 ] such as increasing the membrane area [ 18,19 ] or by forming intracellular lipid droplets which can serve as an intracellular reservoir for hydrophobic compounds. [ 20 ] Although there have been preliminary reports on employing morphology engineering and formation of inner‐membrane vesicles (IMVs) or outer‐membrane vesicles (OMVs) for enhancing the production of hydrophobic compounds, [ 18–23 ] combinatorial applications of these membrane engineering strategies have not been reported. Despite these attempts, the titers of natural colorants need to be further increased for industrial applications.…”

Section: Introductionmentioning

confidence: 99%

Production of Rainbow Colorants by Metabolically Engineered Escherichia coli

2021

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There has been much interest in producing natural colorants to replace synthetic colorants of health concerns. Escherichia coli has been employed to produce natural colorants including carotenoids, indigo, anthocyanins, and violacein. However, production of natural green and navy colorants has not been reported. Many natural products are hydrophobic, which are accumulated inside or on the cell membrane. This causes cell growth limitation and consequently reduces production of target chemicals. Here, integrated membrane engineering strategies are reported for the enhanced production of rainbow colorants-three carotenoids and four violacein derivatives-as representative hydrophobic natural products in E. coli. By integration of systems metabolic engineering, cell morphology engineering, inner-and outer-membrane vesicle formation, and fermentation optimization, production of rainbow colorants are significantly enhanced to 322 mg L -1 of astaxanthin (red), 343 mg L -1 of -carotene (orange), 218 mg L -1 of zeaxanthin (yellow), 1.42 g L -1 of proviolacein (green), 0.844 g L -1 of prodeoxyviolacein (blue), 6.19 g L -1 of violacein (navy), and 11.26 g L -1 of deoxyviolacein (purple). The membrane engineering strategies reported here are generally applicable to microbial production of a broader range of hydrophobic natural products, contributing to food, cosmetic, chemical, and pharmaceutical industries.

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“…Expression of human CAV1 in E. coli induces the formation of heterologous caveolae in the cytoplasmic membrane . The recombinant E. coli can incorporate the periplasmic medium and provide additional membranes for transmembrane protein incorporation. , We hypothesized that endocytosis in E. coli may constitute a general uptake system for biotransformation of substrates, as eukaryotic endocytosis is a form of active transport for incorporating substances from the surrounding medium.…”

Section: Resultsmentioning

confidence: 99%

“…The dye transport assay was performed according to published methods. , CAV1 -expressing cells were cultured in Terrific broth at 37 °C overnight. Precultured cells (1%) were inoculated in Terrific broth and cultured at 37 °C until the optical density (OD) reached 0.3.…”

Section: Methodsmentioning

confidence: 99%

Endocytosing Escherichia coli as a Whole-Cell Biocatalyst of Fatty Acids

Shin

Park

et al. 2019

ACS Synth. Biol.

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Whole cell biocatalysts can be used to convert fatty acids into various value-added products. However, fatty acid transport across cellular membranes into the cytosol of microbial cells limits substrate availability and impairs membrane integrity, which in turn decreases cell viability and bioconversion activity. Because these problems are associated with the mechanism of fatty acid transport through membranes, a wholecell biocatalyst that can form caveolae-like structures was generated to promote substrate endocytosis. Caveolin-1 (CAV1) expression in Escherichia coli increased both the fatty acid transport rate and intracellular fatty acid concentrations via endocytosis of the supplemented substrate. Furthermore, fatty-acid endocytosis alleviated substrate cytotoxicity in E. coli. These traits attributed to bacterial endocytosis resulted in dramatically elevated biotransformation efficiencies in fed-batch and cellrecycle reaction systems when caveolae-forming E. coli was used for the bioconversion of ricinoleic acid (12-hydroxyoctadec-9-enoic acid) to (Z)-11-(heptanoyloxy) undec-9-enoic acid. We propose that CAV1-mediated endocytosing E. coli represents a versatile tool for the biotransformation of hydrophobic substrates.

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Display of membrane proteins on the heterologous caveolae carved by caveolin-1 in the Escherichia coli cytoplasm

Cited by 17 publications

References 31 publications

Ectopic Neo-Formed Intracellular Membranes in Escherichia coli: A Response to Membrane Protein-Induced Stress Involving Membrane Curvature and Domains

Ectopic Neo-Formed Intracellular Membranes in Escherichia coli: A Response to Membrane Protein-Induced Stress Involving Membrane Curvature and Domains

Production of Rainbow Colorants by Metabolically Engineered Escherichia coli

Endocytosing Escherichia coli as a Whole-Cell Biocatalyst of Fatty Acids

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