Increasing the catalytic activity of Bilirubin oxidase from Bacillus pumilus: Importance of host strain and chaperones proteins

Gounel, Sébastien; Rouhana, Jad; Stinès-Chaumeil, Claire; Cadet, Marine; Mano, Nicolas

doi:10.1016/j.jbiotec.2016.04.035

Cited by 17 publications

(21 citation statements)

References 34 publications

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“…BOD from Bacillus pumilus (Bp BOD) was a gift from Dr Nicolas Mano (CRPP, Bordeaux, France). 29 (Table S1). The plasmid pet21b (Addgene, Cambridge, USA) was linearized with primers PRC3 and PRC4 (Table S1).…”

Section: Methodsmentioning

confidence: 99%

Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O₂ Reduction

et al. 2019

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Multicopper oxidases (MCOs) catalyze the oxidation of a variety of substrates while reducing oxygen into water through four copper atoms. As an additional feature, some MCOs display an enhanced activity in solution in the presence of Cu 2+. This is the case of the hyperthermophilic laccase HB27 from Thermus thermophilus, the physiologic role of which is unknown. As a particular feature, this enzyme presents a methionine rich domain proposed to be involved in copper interaction. In this work, laccase from T. thermophilus was produced in E. coli, and the effect of Cu 2+ on its electroactivity at carbon nanotube modified electrodes was investigated. Direct O2 electroreduction is strongly dictated by carbon nanotube surface chemistry in accordance with the enzyme dipole moment. In the presence of Cu 2+ , an additional low potential cathodic wave occurs, which was never described earlier. Analysis of this wave as a function 2 of Cu 2+ availability allows us to attribute this wave to a cuprous oxidase activity displayed by the laccase and induced by copper binding close to the Cu T1 center. A mutant lacking the methionine-rich hairpin domain characteristic of this laccase conserves its copper activity suggesting a different site of copper binding. This study provides new insight into the copper effect in methionine rich MCOs, and highlights the utility of the electrochemical method to investigate cuprous oxidase activity and to understand the physiological role of these MCOs.

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Section: Methodsmentioning

confidence: 99%

Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O₂ Reduction

et al. 2019

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“…To construct the biocathode, two variants of BOD were examined. Thermophilic Bacillus pumilus bilirubin oxidase (BpBOD) was cloned in E. coli by a method described previously with some modifications. We also tested the commonly used BOD that originated from Myrothecium verrucaria (MvBOD).…”

Section: Resultsmentioning

confidence: 99%

“…BOD, a Cu‐based metalloenzyme, can reduce oxygen to water as it oxidizes bilirubin at a neutral pH. This protein is ideal for biotechnological applications because its Cu active site accepts an electron from aromatic moieties, which allows the continuous recycling of atmospheric O 2 . In recent years, photoanodes based on earth‐abundant materials have been developed.…”

Section: Introductionmentioning

confidence: 99%

Bismuth Vanadate/Bilirubin Oxidase Photo(bio)electrochemical Cells for Unbiased, Light‐Triggered Electrical Power Generation

2020

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The construction of bias‐ and donor‐free photobioelectrochemical cells for the generation of light‐triggered electrical power is presented. The developed oxygen reduction biocathodes are based on bilirubin oxidase (BOD) that originates from Myrothecium verrucaria (MvBOD) and a thermophilic Bacillus pumilus (BpBOD). Methods to entrap the BOD with 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) redox molecules in a polydopamine layer are presented. A pH‐independent, positively charged pyrenebetaine linker was synthesized, utilized, and led to a threefold improvement to the bioelectrocatalytic current. Both the developed polydopamine/ABTS/MvBOD and the pyrenebetaine/BpBOD biocathodes were further coupled with BiVO4/cobalt phosphate water‐oxidation photoanodes to construct biotic/abiotic photobioelectrochemical cells, which generated power outputs of 0.74 and 0.85 mW cm−2, respectively. The presented methods are versatile, show the strength of biotic/abiotic hybrids, and can be further used to couple different redox enzymes with electrodes.

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“…The enzyme solution was prepared in a concentration of 20 mg mL −1 in phosphate buffer (pH 7.3). Bilirubin oxidase (BOx) from Bacillus pumilus in 50 mM borate buffer (pH 9) with a protein concentration of 54.75 mg mL −1 (determined in 1 mM 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulphonic acid (ABTS) in 50 mM citrate/phosphate buffer, pH 4) and an activity of 713 units mg −1 was obtained as recently described . The apo‐enzyme of pyrroloquinoline quinone‐dependent glucose dehydrogenase (PQQ‐GDH) form Acinetobacter calcoaceticus (Mut‐Q‐GDH 2 mutant of the native GDH) was a gift from Roche Diagnostics, Penzberg/Germany.…”

Section: Methodsmentioning

confidence: 99%

Introducing Pseudocapacitive Bioelectrodes into a Biofuel Cell/Biosupercapacitor Hybrid Device for Optimized Open Circuit Voltage

et al. 2019

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We report the fabrication of a polymer/enzyme-based biosupercapacitor (BSC)/biofuel cell (BFC) hybrid device with an optimized cell voltage that can be switched on demand from energy conversion to energy storage mode. The redox polymer matrices used for the immobilization of the biocatalyst at the bioanode and biocathode act simultaneously as electron relays between the integrated redox enzymes and the electrode surface (BFC) and as pseudocapacitive charge storing elements (BSC). Moreover, owing to the self-charging effect based on the continuously proceeding enzymatic reaction, a Nernstian shift in the pseudocapacitive elements, that is, in the redox polymers, at the individual bioelectrodes leads to a maximized open circuit voltage of the device in both operating modes. Comparison with a conventional fuel cell design, that is, using redox mediators with redox potentials that are close to the potentials of the used redox proteins, indicates that the novel hybrid device shows a similar voltage output. Moreover, our results demonstrate that the conventional design criteria commonly used for the development of redox polymers for the use in biofuel cells have to be extended by considering the effect of a Nernstian shift towards the potentials of the used biocatalysts in those pseudocapacitive elements.

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Increasing the catalytic activity of Bilirubin oxidase from Bacillus pumilus: Importance of host strain and chaperones proteins

Cited by 17 publications

References 34 publications

Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O₂ Reduction

Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O₂ Reduction

Bismuth Vanadate/Bilirubin Oxidase Photo(bio)electrochemical Cells for Unbiased, Light‐Triggered Electrical Power Generation

Introducing Pseudocapacitive Bioelectrodes into a Biofuel Cell/Biosupercapacitor Hybrid Device for Optimized Open Circuit Voltage

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Increasing the catalytic activity of Bilirubin oxidase from Bacillus pumilus: Importance of host strain and chaperones proteins

Cited by 17 publications

References 34 publications

Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O2 Reduction

Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O2 Reduction

Bismuth Vanadate/Bilirubin Oxidase Photo(bio)electrochemical Cells for Unbiased, Light‐Triggered Electrical Power Generation

Introducing Pseudocapacitive Bioelectrodes into a Biofuel Cell/Biosupercapacitor Hybrid Device for Optimized Open Circuit Voltage

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Product

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Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O₂ Reduction

Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O₂ Reduction