Direct electrical contact of NAD+/NADH-dependent dehydrogenase on electrode surface enabled by non-native solid-binding peptide as a molecular binder

Reginald, Stacy Simai; Kim, Min Ji; Lee, Hyeryeong; Fazil, Nabilah; Choi, Serah; Oh, So-Young; Seo, Junhyeok; Chang, Ilhan

doi:10.1016/j.electacta.2022.140480

Cited by 3 publications

(4 citation statements)

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“…Still, the minor decreases were observed in catalytic activity of fusion enzymes compared with native enzyme, especially when the number of GBP tags are increased. These results can be attributed to partial improper protein folding during DNA translation or structural instability of main catalytic subunit of GBP-fused enzyme . However, over 90% activity retentions of GBP-fused enzymes were achieved compared with those of wild-type enzymes, proving that GBP fusion at the end of enzymatic sequences can facilitate retention of intrinsic enzyme activity without involving conformational changes in active site residues or cofactors.…”

Section: Resultsmentioning

confidence: 99%

“…These results can be attributed to partial improper protein folding during DNA translation or structural instability of main catalytic subunit of GBP-fused enzyme. 38 However, over 90% activity retentions of GBP-fused enzymes were achieved compared with those of wild-type enzymes, proving that GBP fusion at the end of enzymatic sequences can facilitate retention of intrinsic enzyme activity without involving conformational changes in active site residues or cofactors. Moreover, cross-reaction of GDHγα toward sucrose and fructose was observed because they could coexist with GDHγα during cascade catalysis of coimmobilized bienzymes.…”

Section: Catalytic Activities Of Native and Fusionmentioning

confidence: 99%

“…The use of solid-binding peptides (SBPs) as molecular linkers is a plausible approach as an orienting mediator for enzyme-electrode or enzyme–enzyme orientation. , SBPs are short amino acid sequences that possess a high affinity for recognizing certain inorganic materials. , Thus, the genetic linkage of SBP fragment amino acid sequences at the terminus of an enzymatic structure can specify the anchoring orientation of the fusion enzyme depending on the linkage site. Moreover, the concurrent surface-orientation control of coupling enzymes on the electrode results in their varied relative interenzyme orientation.…”

Section: Introductionmentioning

confidence: 99%

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Orientation-Controllable Enzyme Cascade on Electrode for Bioelectrocatalytic Chain Reaction

Lee

Bang

Chang

2023

ACS Appl. Mater. Interfaces

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The accomplishment of concurrent interenzyme chain reaction and direct electric communication in a multienzyme-electrode is challenging since the required condition of multienzymatic binding conformation is quite complex. In this study, an enzyme cascade-induced bioelectrocatalytic system has been constructed using solid binding peptide (SBP) as a molecular binder that coimmobilizes the invertase (INV) and flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenase gamma-alpha complex (GDHγα) cascade system on a single electrode surface. The SBP-fused enzyme cascade was strategically designed to induce diverse relative orientations of coupling enzymes while enabling efficient direct electron transfer (DET) at the FAD cofactor of GDHγα and the electrode interface. The interenzyme relative orientation was found to determine the intermediate delivery route and affect overall chain reaction efficiency. Moreover, interfacial DET between the fusion GDHγα and the electrode was altered by the binding conformation of the coimmobilized enzyme and fusion INVs. Collectively, this work emphasizes the importance of interenzyme orientation when incorporating enzymatic cascade in an electrocatalytic system and demonstrates the efficacy of SBP fusion technology as a generic tool for developing cascade-induced direct bioelectrocatalytic systems. The proposed approach is applicable to enzyme cascade-based bioelectronics such as biofuel cells, biosensors, and bioeletrosynthetic systems utilizing or producing complex biomolecules.

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Section: Resultsmentioning

confidence: 99%

Section: Catalytic Activities Of Native and Fusionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Orientation-Controllable Enzyme Cascade on Electrode for Bioelectrocatalytic Chain Reaction

Lee

Bang

Chang

2023

ACS Appl. Mater. Interfaces

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“…Effects of design factors on performance parameters References SBP fusion site Catalytic activity ✓ Catalytic activity of enzyme tends to be highly preserved after genetic SBP fusion. [29,34,44] Inorganic-binding activity ✓ SBP fusion to multiple sites can increase overa ll inorganic-binding affinities. ✓ The external exposure degree of fusion site (i.e., C-or N-terminus of protein structure) can determine contact probability of SBP domain with target surfaces.…”

Section: Design Factors Performance Parametersmentioning

confidence: 99%

Versatile Biosynthetic Approach to Regioselective Enzyme Patterning for Direct Bioelectrocatalysis Application

Lee

Reginald

Chang

2023

Adv Materials Technologies

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accomplish enzymatic catalysis with high catalytic efficiency. [6,7] Specifically, such a spatial organization of enzymes enables the efficient delivery of metabolites among enzyme active sites while preventing the steric hindrance of a reaction site by adjacent proteins. [8] Therefore, it is essential to consider the effect of enzyme spatial organization on overall metabolic efficiency for translating our knowledge about natural biochemical systems into noncellular artificial environments, such as those involving electrodes. [9] Inspired by nature, a generation of proper multior single enzymatic positioning on electrodes can bring about enhanced performance of enzyme-based bioelectrocatalysis systems. [10] Regarding in vitro spatial organization, there have been considerable efforts to achieve technical advances in position-specific enzyme immobilization by performing "protein patterning". [11,12] Protein molecules have been patterned using direct or indirect assembly (i.e., self-assembled monolayers alkanethiol, silane, Ni-NTA, biotin, etc.) by integration with lithographic approaches including scanning probe microscopy, [13,14] dip-pen nanolithography (DPN), [15] atomic force microscopy (AFM), [16] electron beam (e-beam) lithography (EBL), [17] and nanoimprint lithography (NIL). [18] DNA templates have also been widely used to create periodic arrays of protein molecules. [19] Yan et al. (2003) reported that DNA tiles with sticky ends self-assembled into nanoribbons or nanogrids that provided a predetermined surface for target-molecule binding. [20] Since the enzyme-based electrocatalysis system operates cooperatively by single-or multienzyme-based catalysis and electron transfer (ET) between the enzymatic cofactor and electrode, electrical communication of the enzyme-electrode should also be considered and controlled together with enzymatic surface-positioning. [21,22] However, most of the previous studies involving the development of protein micro/nanopatterning techniques were not primarily concerned with the enzymeelectrode interface. The approaches for protein patterning have not proven to be amenable in controlling the orientations of enzymes on surfaces. Hence, extensive chemical/biochemical surface modification or random attachments of the protein may electrically block the cofactor-surface interfaces, limiting the interfacial efficiency of direct ET (DET). [23] Due to the outstanding attributes of oxidoreductases, they have been utilized as biomaterials for bioelectrocatalytic systems. Herein, a simple and versatile biosynthetic approach that can designate binding position of enzymes on electrode with their surface-orientation is suggested. In this regard, materialselective properties of gold-binding peptide (GBP) are exploited and genetically fused GBP to enzyme. To optimize the design of synthetic enzyme, a variable repeat number of GBP are fused to flavin adenine dinucleotidedependent glucose dehydrogenase gamma-alpha complex (GDHγα) and their catalytic and gold-binding activities are determined. T...

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Advanced manufacture of biofuel cells

Zhao,

Hao,

Zhang

et al. 2024

Biofuel Cells

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Direct electrical contact of NAD+/NADH-dependent dehydrogenase on electrode surface enabled by non-native solid-binding peptide as a molecular binder

Cited by 3 publications

References 54 publications

Orientation-Controllable Enzyme Cascade on Electrode for Bioelectrocatalytic Chain Reaction

Orientation-Controllable Enzyme Cascade on Electrode for Bioelectrocatalytic Chain Reaction

Versatile Biosynthetic Approach to Regioselective Enzyme Patterning for Direct Bioelectrocatalysis Application

Advanced manufacture of biofuel cells

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