Bioluminescence of Pseudomonas fluorescens HK44 in the course of encapsulation into silica gel. Effect of methanol

Trögl, Josef; Kuncová, Gabriela; Kuráň, Pavel

doi:10.1007/s12223-010-0091-9

Cited by 10 publications

(12 citation statements)

References 40 publications

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“…The main disadvantages of this biosensor configuration were the length of time needed for preparation of the cell layer and continual changes of analytical responses. In this work, we applied an optimized route toward [19,29,30,34] silica-gel-encapsulation of living bioreporters on to optical fibers. In contrast to physical adsorption, this technique enables reproducible fixation of a desired amount of cells on the top of the optical fiber within an hour [35].…”

Section: Resultsmentioning

confidence: 99%

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The Repetitive Detection of Toluene with Bioluminescence Bioreporter Pseudomonas putida TVA8 Encapsulated in Silica Hydrogel on an Optical Fiber

et al. 2016

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Living cells of the lux-based bioluminescent bioreporter Pseudomonas putida TVA8 were encapsulated in a silica hydrogel attached to the distal wider end of a tapered quartz fiber. Bioluminescence of immobilized cells was induced with toluene at high (26.5 mg/L) and low (5.3 mg/L) concentrations. Initial bioluminescence maxima were achieved after >12 h. One week after immobilization, a biofilm-like layer of cells had formed on the surface of the silica gel. This resulted in shorter response times and more intensive bioluminescence maxima that appeared as rapidly as 2 h after toluene induction. Considerable second bioluminescence maxima were observed after inductions with 26.5 mg toluene/L. The second and third week after immobilization the biosensor repetitively and semiquantitatively detected toluene in buffered medium. Due to silica gel dissolution and biofilm detachment, the bioluminescent signal was decreasing 20–32 days after immobilization and completely extinguished after 32 days. The reproducible formation of a surface cell layer on the wider end of the tapered optical fiber can be translated to various whole cell bioluminescent biosensor devices and may serve as a platform for in-situ sensors.

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Section: Resultsmentioning

confidence: 99%

“…Cell encapsulation was performed according to a procedure used in previous studies [19,29,30,31]. Briefly, tetramethoxysilane (TMOS, 4.1 g) was stirred with distilled water (2 mL), cooled for 5 min at 4 °C, and 0.1 M HCl (0.5 mL) was then slowly added.…”

Section: Methodsmentioning

confidence: 99%

The Repetitive Detection of Toluene with Bioluminescence Bioreporter Pseudomonas putida TVA8 Encapsulated in Silica Hydrogel on an Optical Fiber

et al. 2016

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“…An exponentially growing HK44 culture emits higher non-induced (background) bioluminescence and hence the relative increase upon induction is lower as compared to stationary or starving cultures [48,54]. Bioluminescence also decreases upon interaction with toxic chemicals, including inducers of nah-lux genes in strain HK44 [54–57]. It was also demonstrated that HK44 bioluminescence increases upon exposure to organic solvents [54,58].…”

Section: Biosensing With P Fluorescens Hk44mentioning

confidence: 99%

“…This effect is known from other bioluminescent assays [59] and is ascribed to membrane perturbations which, through production of increased levels of free fatty acids, leads to an increased supply of aldehyde substrates for luciferase that are normally limited, although this remains hypothetical. At high concentrations of perturbing chemicals, the negative effect prevails leading to a decrease in bioluminescence (Figure 5) [54–57]. …”

Section: Biosensing With P Fluorescens Hk44mentioning

confidence: 99%

“…However, the fundamental knowledge regarding immobilized cell physiology is somewhat lagging behind the applied research [38,84]. Bioluminescent microorganisms provide a simple method for monitoring the viability and physiological stressors that impact cells under immobilized states, with strain HK44 being applied in several such studies [52,57,70]. …”

Section: Biosensing With P Fluorescens Hk44mentioning

confidence: 99%

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Pseudomonas fluorescens HK44: Lessons Learned from a Model Whole-Cell Bioreporter with a Broad Application History

Trögl

Chauhan

Ripp

et al. 2012

Sensors

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Initially described in 1990, Pseudomonas fluorescens HK44 served as the first whole-cell bioreporter genetically endowed with a bioluminescent (luxCDABE) phenotype directly linked to a catabolic (naphthalene degradative) pathway. HK44 was the first genetically engineered microorganism to be released in the field to monitor bioremediation potential. Subsequent to that release, strain HK44 had been introduced into other solids (soils, sands), liquid (water, wastewater), and volatile environments. In these matrices, it has functioned as one of the best characterized chemically-responsive environmental bioreporters and as a model organism for understanding bacterial colonization and transport, cell immobilization strategies, and the kinetics of cellular bioluminescent emission. This review summarizes the characteristics of P. fluorescens HK44 and the extensive range of its applications with special focus on the monitoring of bioremediation processes and biosensing of environmental pollution.

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Engineering of a silica encapsulation platform for hydrocarbon degradation using Pseudomonas sp. NCIB 9816‐4

Sakkos

Kieffer

Mutlu

et al. 2015

Biotech & Bioengineering

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Industrial application of encapsulated bacteria for biodegradation of hydrocarbons in water requires mechanically stable materials. A silica gel encapsulation method was optimized for Pseudomonas sp. NCIB 9816-4, a bacterium that degrades more than 100 aromatic hydrocarbons. The design process focused on three aspects: (i) mechanical property enhancement; (ii) gel cytocompatibility; and (iii) reduction of the diffusion barrier in the gel. Mechanical testing indicated that the compressive strength at failure (σf ) and elastic modulus (E) changed linearly with the amount of silicon alkoxide used in the gel composition. Measurement of naphthalene biodegradation by encapsulated cells indicated that the gel maintained cytocompatibility at lower levels of alkoxide. However, significant loss in activity was observed due to methanol formation during hydrolysis at high alkoxide concentrations, as measured by FTIR spectroscopy. The silica gel with the highest amount of alkoxide (without toxicity from methanol) had a biodegradation rate of 285 ± 42 nmol/L-s, σf = 652 ± 88 kPa, and E = 15.8 ± 2.0 MPa. Biodegradation was sustained for 1 month before it dropped below 20% of the initial rate. In order to improve the diffusion through the gel, polyvinyl alcohol (PVA) was used as a porogen and resulted in a 48 ± 19% enhancement in biodegradation, but it impacted the mechanical properties negatively. This is the first report studying how the silica composition affects biodegradation of naphthalene by Pseudomonas sp. NCIB 9816-4 and establishes a foundation for future studies of aromatic hydrocarbon biodegradation for industrial application.

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Bioluminescence of Pseudomonas fluorescens HK44 in the course of encapsulation into silica gel. Effect of methanol

Cited by 10 publications

References 40 publications

The Repetitive Detection of Toluene with Bioluminescence Bioreporter Pseudomonas putida TVA8 Encapsulated in Silica Hydrogel on an Optical Fiber

The Repetitive Detection of Toluene with Bioluminescence Bioreporter Pseudomonas putida TVA8 Encapsulated in Silica Hydrogel on an Optical Fiber

Pseudomonas fluorescens HK44: Lessons Learned from a Model Whole-Cell Bioreporter with a Broad Application History

Engineering of a silica encapsulation platform for hydrocarbon degradation using Pseudomonas sp. NCIB 9816‐4

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