A patch coating method for preparing biocatalytic films ofEscherichia coli

Lyngberg, Olav; Thiagarajan, V.; Stemke, D. J.; Schottel, Janet L.; Scriven, L. E.; Flickinger, Michael C.

doi:10.1002/(sici)1097-0290(19990105)62:1<44::aid-bit6>3.0.co;2-w

Cited by 35 publications

(45 citation statements)

References 18 publications

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“…Such a situation may be akin to that of the polymer-encased bacteria that are used for biocatalytic engineering applications (25). Although these bacteria do not divide, they are viable and culturable once freed from the plastic encasement.…”

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confidence: 99%

Biofilm, City of Microbes

2000

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confidence: 99%

Biofilm, City of Microbes

2000

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“…Cells were harvested at 6k rpm at 4°C for 10 min and washed twice in phosphate-buVered saline (PBS), pH 7.4 [1]. To prepare the latex-immobilized cell patches, the procedure of Lyngberg et al [23] was used with modiWcations as follows. The cell pellet was weighed, and 0.6 g of cell paste was resuspended in a 150 l solution containing 75 l of 1.3 g/ml (wt/v) glycerol, 10 l of 1% (v/v) Tween 20 (to prevent aggregation of the microspheres), 50 l Flash Red Xuorescent carboxylate-modiWed microspheres (0.19 m diameter, 1% wt/v, 660/690 nm excitation/emission; Bangs Laboratories, Fishers, IN, USA), and 15 l sterile distilled water.…”

Section: Cell Immobilization In Latexmentioning

confidence: 99%

“…The Wnal cell-latex mixture contained approximately 50% (wt/v) of E. coli cells, 7.8% (wt/v) glycerol, 0.008% (v/v) Tween 20, and 0.04% (wt/v) Flash Company, Pompano Beach, FL, USA). The cell coat was dried at room temperature (21-24°C) at 50 § 5% relative humidity (RH) for 1 h. The dried cell coat was about 20-m thick, and each patch contained approximately 5 £ 10 8 cells [23].…”

Section: Cell Immobilization In Latexmentioning

confidence: 99%

“…As opposed to immobilized enzymes, living cells can generate the expensive but critical reaction components such as ATP and NADPH, but the immobilization process must maintain cell viability and reactivity for the desired period of time. Adhesive latex has been used as an immobilization support for the development of biosensors and biocatalysts containing whole cells [8][9][10] and is available in a variety of polymer particle sizes and formulations with diVerent coalescence characteristics [20][21][22][23]. These coatings are typically composed of one or more cell coat layers in which the cells are mixed with latex and spread as a 10-to 70-m thick sheet onto a polyester or metal substrate.…”

Section: Introductionmentioning

confidence: 99%

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Spatial expression of a mercury-inducible green fluorescent protein within a nanoporous latex-based biosensor coating

Schottel

Orwin

Anderson

et al. 2008

J Ind Microbiol Biotechnol

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Optimizing the reactivity of cell coatings developed as biosensors or biocatalysts requires measurements of gene expression in the immobilized cells. To quantify and localize gene expression within a latex-based mercury biosensor, a plasmid, pmerGFP, was constructed, which contains the green fluorescent protein (GFP) gene under transcriptional control of the mercury resistance operon regulatory sequences. When cells containing this plasmid were exposed to mercuric chloride, GFP synthesis was induced and could be quantified by fluorescence. E. coli strain JM109 (pmerGFP) was mixed with SF091 latex (Rohm & Haas), Tween 20, and glycerol, and coated as an approximate 20-microm thick nanoporous adhesive coating on a polyester substrate. The cell coat was overlaid with a nanoporous topcoat of latex, Tween 20, and glycerol. Different fluorescent microspheres were used to mark the topcoat and cell coat layers of the coating. Upon exposure to mercury(II), cells within the coating were induced to synthesize GFP, and laser scanning confocal microscopy was used to quantify expression spatially within the cell coat. GFP expression in the coatings increased with increasing mercury concentration (2-20 microM), temperature (21-37 degrees C), and time of incubation (0-39 h). There was a gradient of GFP expression through the cell coat with expression higher near the topcoat-cell coat interface relative to the bottom of the cell coat. The topcoat thickness did not significantly affect GFP expression indicating that diffusion of mercury(II) and oxygen through the topcoat was not limiting.

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“…Nanoporous, adhesive latex biocatalytic coatings offer the possibility of concentrating and entrapping viable microorganisms (such as bacteria, yeast or fungi) in very thin (five to <75 µm ), partially-coalesced polymer coatings [9][10][11][12][13][14][15][16][17][18][19][20]. Their thinness, engineered adhesion, nanoporous microstructure, mass-transfer properties [21][22][23], high-entrapped cell density and ability to be stored partially desiccated at ambient temperature differ significantly from the entrapment of viable microorganisms in crosslinked micro-porous carbohydrate gels, ceramic matrices, monoliths, soft synthetic polymer matrices (such as alginate or polyacrylamide) or behind membranes.…”

Section: Introductionmentioning

confidence: 99%

Ink-Jet Printing of Gluconobacter oxydans: Micropatterned Coatings As High Surface-to-Volume Ratio Bio-Reactive Coatings

et al. 2013

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Abstract:We formulated a latex ink for ink-jet deposition of viable Gram-negative bacterium Gluconobacter oxydans as a model adhesive, thin, highly bio-reactive microstructured microbial coating. Control of G. oxydans latex-based ink viscosity by dilution with water allowed ink-jet piezoelectric droplet deposition of 30 × 30 arrays of two or three droplets/dot microstructures on a polyester substrate. Profilometry analysis was used to study the resulting dry microstructures. Arrays of individual dots with base diameters of ~233-241 µm were obtained. Ring-shaped dots with dot edges higher than the center, 2.2 and 0.9 µm respectively, were obtained when a one-to-four diluted ink was used. With a less diluted ink (one-to-two diluted), the microstructure became more uniform with an average height of 3.0 µm , but the ink-jet printability was more difficult. Reactivity of the ink-jet deposited microstructures following drying and rehydration was studied in a non-growth medium by oxidation of 50 g/L D-sorbitol to L-sorbose, and a high dot volumetric reaction rate was measured (~435 g· L −1 · h −1 ). These results indicate that latex ink microstructures generated by ink-jet printing may hold considerable potential for 3D fabrication of high surface-to-volume ratio biocoatings for use as microbial biosensors with the aim of coating microbes as reactive biosensors on electronic devices and circuit chips. OPEN ACCESSCoatings 2014, 4 2

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A patch coating method for preparing biocatalytic films ofEscherichia coli

Cited by 35 publications

References 18 publications

Biofilm, City of Microbes

Biofilm, City of Microbes

Spatial expression of a mercury-inducible green fluorescent protein within a nanoporous latex-based biosensor coating

Ink-Jet Printing of Gluconobacter oxydans: Micropatterned Coatings As High Surface-to-Volume Ratio Bio-Reactive Coatings

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