Versatile bioelectronic interfaces based on heterotrifunctional linking molecules

Hassler, Brian L.; Worden, R. Mark

doi:10.1016/j.bios.2005.10.004

Cited by 24 publications

(12 citation statements)

References 39 publications

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“…Improved experimental tools are needed that maintain membrane proteins in an environment that allows their activities to be simultaneously expressed and measured in vitro [1]. Such tools could be used in functional proteomics research to characterize novel proteins, in high-throughput drug screening to investigate receptor-drug interactions, and in development of novel biosensor systems that exploit specific interactions between proteins and analytes [2,3].…”

Section: Introductionmentioning

confidence: 99%

Fabrication of highly insulating tethered bilayer lipid membrane using yeast cell membrane fractions for measuring ion channel activity

Jadhav

Sui

Garavito

et al. 2008

Journal of Colloid and Interface Science

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Section: Introductionmentioning

confidence: 99%

Fabrication of highly insulating tethered bilayer lipid membrane using yeast cell membrane fractions for measuring ion channel activity

Jadhav

Sui

Garavito

et al. 2008

Journal of Colloid and Interface Science

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“…At 400 mV, nicotinamide adenine dinucleotide phosphate (NADPH) is oxidized in the presence of TBO [29]. Liquid samples were analyzed for 2-propanol and acetone using a JEOL AX505H double-focusing mass spectrometer (MS) (Japan Electronic Optics Laboratories, Peabody, MA, USA) equipped with a Hewlett-Packard 6890J gas chromatograph (GC) (Palo Alto, CA, USA) and a 30 DB1 fused silica column.…”

Section: Measurement Of Catalytic Activitymentioning

confidence: 99%

“…We recently developed an electron-transfer scaffold using a heterotrifunctional linking molecule (e.g., cysteine, CYS) [29]. The electron mediator and cofactor were bound to opposite branches of the linking molecule, allowing greater flexibility in scaffold formation and permitting the use of electron mediators having a single reactive group.…”

mentioning

confidence: 99%

Mutation of Tyr-218 to Phe in Thermoanaerobacter ethanolicus Secondary Alcohol Dehydrogenase: Effects on Bioelectronic Interface Performance

Hassler

Dennis

Laivenieks

et al. 2007

Appl Biochem Biotechnol

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Bioelectronic interfaces that facilitate electron transfer between the electrode and a dehydrogenase enzyme have potential applications in biosensors, biocatalytic reactors, and biological fuel cells. The secondary alcohol dehydrogenase (2 degrees ADH) from Thermoanaerobacter ethanolicus is especially well suited for the development of such bioelectronic interfaces because of its thermostability and facile production and purification. However, the natural cofactor for the enzyme, beta-nicotinamide adenine dinucleotide phosphate (NADP+), is more expensive and less stable than beta-nicotinamide adenine dinucleotide (NAD+). PCR-based, site-directed mutagenesis was performed on 2 degrees ADH in an attempt to adjust the cofactor specificity toward NAD+ by mutating Tyr218 to Phe (Y218F 2 degrees ADH). This mutation increased the Km(app) for NADP+ 200-fold while decreasing the Km(app) for NAD+ 2.5-fold. The mutant enzyme was incorporated into a bioelectronic interface that established electrical communication between the enzyme, the NAD+, the electron mediator toluidine blue O (TBO), and a gold electrode. Cyclic voltammetry, impedance spectroscopy, gas chromatography, mass spectrometry, constant potential amperometry, and chronoamperometry were used to characterize the mutant and wild-type enzyme incorporated in the bioelectronic interface. The Y218F 2 degrees ADH exhibited a fourfold increase in the turnover ratio compared to the wild type in the presence of NAD+. The electrochemical and kinetic measurements support the prediction that the Rossmann fold of the enzyme binds to the phosphate moiety of the cofactor. During the 45 min of continuous operation, NAD+ was electrically recycled 6.7 x 10(4) times, suggesting that the Y218F 2 degrees ADH-modified bioelectronic interface is stable.

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“…(1)(2)(3)(4) This is because H 2 O 2 is the main component of reactive oxygen species (ROS) in living organisms, and increasing levels of H 2 O 2 in cells could indicate oxidative stress and cellular damage. (5) This damage could result in many diseases such as cancer, neurodegenerative disorders, diabetes, and atherosclerosis.…”

Section: Introductionmentioning

confidence: 99%

Improvement in Sensing Performance of H2O2 Biosensor Electrodes through Modification of Anatase TiO2 Nanorods and Pretreatment of Electrochemical Reduction

2017

Sensors and Materials

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Electrochemical biosensors are essential health monitors that aid in the detection and diagnosis of diseases. In this research, anatase titanium dioxide (TiO 2 ) nanorods (TNR-A) were synthesized on a titanium (Ti) substrate in three stages, namely, hydrothermal, alkali, and heat treatments, and utilized as a modified electrode (TNR-A/Ti). The pretreatment of electrochemical reduction was adopted to increase the Ti 3+ amount in TiO 2 nanorod surfaces of the electrodes, thus improving electron transfer. The electrodes were characterized by X-ray diffractometry (XRD), fieldemission scanning electron microscopy (FESEM), and X-ray photoelectron spectroscopy (XPS). The electrodes after pretreatment (Ti 3+ -TNR-A/Ti) showed a better electrochemical response to hydrogen peroxide (H 2 O 2 ), indicating that pretreatment improves the performance of electrodes. The assembled biosensor electrode (Nafion/HRP/Ti 3+ -TNR-A/Ti) exhibited a sensitivity of 6096.4 µA•mM −1 •cm −2 , a detection limit of 0.008 µM, a linearity of 0.04-700 µM, and an apparent Michaelis-Menten constant K app M of 0.0027 µM, which are higher than those in previous research studies on TiO 2 or similarly nanostructured modified biosensor electrodes. This research could provide a potential competent method that can be used to modify electrodes for high-performance amperometric biosensors.

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Versatile bioelectronic interfaces based on heterotrifunctional linking molecules

Cited by 24 publications

References 39 publications

Fabrication of highly insulating tethered bilayer lipid membrane using yeast cell membrane fractions for measuring ion channel activity

Fabrication of highly insulating tethered bilayer lipid membrane using yeast cell membrane fractions for measuring ion channel activity

Mutation of Tyr-218 to Phe in Thermoanaerobacter ethanolicus Secondary Alcohol Dehydrogenase: Effects on Bioelectronic Interface Performance

Improvement in Sensing Performance of H2O2 Biosensor Electrodes through Modification of Anatase TiO2 Nanorods and Pretreatment of Electrochemical Reduction

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