Directed evolution of a 6-phosphogluconate dehydrogenase for operating an enzymatic fuel cell at lowered anodic pHs

Ma, Chunling; Wu, Ranran; Huang, Rui; Jiang, Wenxia; You, Chun; Zhu, Leilei; Zhu, Zhiguang

doi:10.1016/j.jelechem.2019.113444

Cited by 10 publications

(13 citation statements)

References 42 publications

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“…So far, there have been tremendous efforts focusing on improving the performance of EBFCs (e.g., power output and lifetime) and broadening their applications, such as constructing novel 3-dimensional nanomaterials for bioelectrodes, utilizing highly stable enzymes and biomimetic cofactors, as well as optimizing electrolyte composition and concentration (Campbell et al 2012;Chen et al 2016;Sakai et al 2009). In our previous study, we demonstrated that lowering anodic pH may be a useful strategy for constructing high-performance in EBFCs, as the proton concentration at the lowered pH can be significantly increased at the anode and it therefore provided a high proton transport driving force (Ma et al 2019). However, the DI used was derived from G. stearothermophilus with an optimum pH at 7.3 and still suffered from the poor activity at low pHs, severely limiting the long-term operation of the EBFC.…”

Section: Introductionmentioning

confidence: 93%

“…The double-layer-based TNBT screening method was carried out as previously described with some modifications ( Fig. 1a) (Ma et al 2019). Transformed cells containing mutant plasmid libraries were firstly incubated on the LB kanamycin agar plate at 37 °C for 16 h. Then, the colonies were treated at 70 °C for 1 h to lyse the cell.…”

Section: Screening Of the Mutants With High Activity At Lowered Phsmentioning

confidence: 99%

“…Recently, a DI has been engineered to increase its reactivity against a specific mediator, 2-amino-1,4-naphthoquinone, aiming to improve the power output of the EBFC (Sugiyama et al 2010). A 6PGDH has been directed evolved toward increased acid-tolerance so that the operation conditions of the EBFC can be broadened (Ma et al 2019).…”

Section: Introductionmentioning

confidence: 99%

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Engineering a diaphorase via directed evolution for enzymatic biofuel cell application

Liu

You

et al. 2020

Bioresour. Bioprocess.

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Background: Diaphorase (DI) has received wide attention as the key anodic enzyme mediating the electron transfer and electric energy generation in enzymatic biofuel cells (EBFCs). Lowering the anodic pH may be a useful strategy for constructing high-performance in EBFCs. However, most DI suffered from the poor activity at low pHs. Therefore, it is necessary to modify the activity and its acidic tolerance to further improve the performance of the EBFC. Results: This paper attempts to improve the enzyme activity of DI originated from Geobacillus stearothermophilus under acidic conditions through directed evolution. Three rounds of random mutagenesis by error-prone PCR of the GsDI gene followed by high-throughput screening allowed the identification of the mutant 3-8 (H37Q, S73T, F105L, S68T, G61S, D74V) exhibiting a 4-or 7-fold increase in the catalytic activity at pH 5.4 or 4.5 compared to that of the wild type. And the pH stability of mutant 3-8 was significantly better than that of wild type and showed a 1.3 times higher in the stability at pH 5.4. The EBFC anode equipped with 0.5 mg of mutant 3-8 achieved a maximum current of 40 μA at pH 5.4, much higher than that with the same loading of the wild type enzyme. Conclusion: The GsDI has been improved in the specific activity and pH stability by directed evolution which leads to the improvement of the EBFC performance. Also, the enlarged catalytic channel of mutant and decreased B-factor may be beneficial for the activity and stability. These results suggest that this engineered DI will be a useful candidate for the construction of enhanced EBFCs.

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Section: Introductionmentioning

confidence: 93%

Section: Screening Of the Mutants With High Activity At Lowered Phsmentioning

confidence: 99%

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Engineering a diaphorase via directed evolution for enzymatic biofuel cell application

Liu

You

et al. 2020

Bioresour. Bioprocess.

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“…A convenient immobilization method recommended for interaction between enzyme and support [30] needs to increase the enzyme's storage, solvent, pH, and thermal stability [3,33,38]. However, some factors have been detected behind enzyme-support interactions that lead to low power and energy density.…”

Section: The Enzyme Support and Substrate In Ebfcmentioning

confidence: 99%

“…The high porosity and low cost of this material increase the active site for the enzyme and give 1.37 times higher power density than without CNTs. A convenient immobilization method recommended for interaction between e and support [30] needs to increase the enzyme's storage, solvent, pH, and thermal s [3,33,38]. However, some factors have been detected behind enzyme-support inter that lead to low power and energy density.…”

Section: The Enzyme Support and Substrate In Ebfcmentioning

confidence: 99%

Mini-Review: Recent Technologies of Electrode and System in the Enzymatic Biofuel Cell (EBFC)

Karim

Yang

2021

Applied Sciences

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Enzymatic biofuel cells (EBFCs) is one of the branches of fuel cells that can provide high potential for various applications. However, EBFC has challenges in improving the performance power output. Exploring electrode materials is one way to increase enzyme utilization and lead to a high conversion rate so that efficient enzyme loading on the electrode surface can function correctly. This paper briefly presents recent technologies developed to improve bio-catalytic properties, biocompatibility, biodegradability, implantability, and mechanical flexibility in EBFCs. Among the combinations of materials that can be studied and are interesting because of their properties, there are various nanoparticles, carbon-based materials, and conductive polymers; all three have the advantages of chemical stability and enhanced electron transfer. The methods to immobilize enzymes, and support and substrate issues are also covered in this paper. In addition, the EBFC system is also explored and developed as suitable for applications such as self-pumping and microfluidic EBFC.

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Enzyme engineering and its industrial applications

Amatto

Rosa-Garzon

Simões

et al. 2021

Biotech and App Biochem

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Recently, there has been an increase in the demand for enzymes with modified activity, specificity, and stability. Enzyme engineering is an important tool to meet the demand for enzymes adjusted to different industrial processes. Knowledge of the structure and function of enzymes guides the choice of the best strategy for engineering enzymes. Each enzyme engineering strategy, such as rational design, directed evolution, and semi-rational design, has specific applications, as well as limitations, which must be considered when choosing a suitable strategy. Engineered enzymes can be optimized for different industrial applications by choosing the appropriate strategy. This review features engineered enzymes that have been applied in food, animal feed, pharmaceuticals, medical applications, bioremediation, biofuels, and detergents.

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Directed evolution of a 6-phosphogluconate dehydrogenase for operating an enzymatic fuel cell at lowered anodic pHs

Cited by 10 publications

References 42 publications

Engineering a diaphorase via directed evolution for enzymatic biofuel cell application

Engineering a diaphorase via directed evolution for enzymatic biofuel cell application

Mini-Review: Recent Technologies of Electrode and System in the Enzymatic Biofuel Cell (EBFC)

Enzyme engineering and its industrial applications

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