Covalent interlocking of glucose oxidase and peroxidase in the voids of paper: enzyme–polymer “spider webs”

Riccardi, Caterina; Mistri, Dipali; Hart, Owen; Anuganti, Murali; Lin, Yao; Kasi, Rajeswari M.; Kumar, Challa V.

doi:10.1039/c6cc00037a

Cited by 32 publications

(22 citation statements)

References 27 publications

(24 reference statements)

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“…Color appeared after 5 min, and the darkness of the color was qualitatively proportional to the concentration of glucose (Figure C). The response could be quantitated using a scanner and image analysis software such as ImageJ per a previously reported method . In this manner, a simple method for visual detection of glucose without the need for fluorescence spectroscopy was also demonstrated using the BSA hydrogel.…”

Section: Resultsmentioning

confidence: 98%

Protein Biophosphors: Biodegradable, Multifunctional, Protein‐Based Hydrogel for White Emission, Sensing, and pH Detection

Benson

Ghimire

Pattammattel

et al. 2017

Adv Funct Materials

108

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A highly efficient, multifunctional, bioderived white-emitting hydrogel (biophosphor) consisting of crosslinked bovine serum albumin and three fluorescent dyes, Coumarin 460, fluorescein, and 5(6)-carboxy-x-rhodamine, is reported here. White emission is obtained upon excitation of the biophosphor at 365 nm with appropriate mole ratios of the above dyes. The CIE 1931 chromaticity coordinates of white emission with 365 nm excitation are (0.36, 0.37), and the correlated color temperature is 5300 K. Multifunctional nature of the biophosphor is also demonstrated. A UV-light-emitting-diode (361 nm) coated with this biophosphor, for example, indicates white emission (CIE 0.28, 0.31) with a half-life of 106 (±5) h. The white emission is also highly sensitive to pH over a broad range (pH 1-11). Incorporation of glucose oxidase and peroxidase in the biophosphor allows for the detection of glucose over a physiologically relevant range of 1.8-288 mg dL −1 . This is a unique, advanced biophosphor with LED and sensing applications, and it is the first example of a multifunctional, proteinaceous white emitter. molar absorptivity and quantum efficiency of emissive components is a significant challenge for systems that combine direct emission and sensitized emission via Förster resonance energy transfer (FRET) to produce white light. [6,7] Hydrogels are hydrophilic polymer networks [8] and are emerging as versatile new matrices for high-efficiency generation of white light using intermolecular energy transfer processes. The gel matrix improves energy transfer efficiency by rigidifying the orientation of the donor (D)-acceptor (A) pairs [9] and preventing their aggregation, which can lead to quenching. [10] Despite the advantageous properties of white-emitting hydrogels, implementation in a functional device and photostability were not systematically studied, and biodegradability has not been demonstrated. Additionally, white-emitting proteinbased hydrogels are not known other than a report of a gelatin hydrogel with chromaticity coordinates [0.26, 0.33], far from being coordinates of pure white emission [0.33, 0.33]. [2] To the best of our knowledge, there are no reports of a multifunctional, nontoxic, biodegradable, white-emitting protein hydrogel.BSA is inexpensive and readily available as a waste product of the meat industry. BSA has a large number of primary amines (59 lysine) and carboxylic acids (99 aspartic acid/glutamic acid), [11] which can be crosslinked under controlled conditions by carbodiimide chemistry to form a network of amide bonds without disrupting the intricate secondary structure of the protein. [12,13] The protein's secondary structure plays an important role for dye binding at the intended site and for enzyme activity retention, when enzymes are incorporated in the matrix for sensing or catalytic applications. We envisioned that this molecular network of BSA would result in a water-rich hydrogel with discrete sites for dye binding which would be suitable for the construction of a white-emitting gel.Previou...

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Section: Resultsmentioning

confidence: 98%

Protein Biophosphors: Biodegradable, Multifunctional, Protein‐Based Hydrogel for White Emission, Sensing, and pH Detection

Benson

Ghimire

Pattammattel

et al. 2017

Adv Funct Materials

108

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“…[4] This method however, is not suitable for enzymes with insufficient lysine residues to attach to the carboxylic acid groups of PA A, or for enzymes inactivated by PA Ao r cellulose. [4] This method however, is not suitable for enzymes with insufficient lysine residues to attach to the carboxylic acid groups of PA A, or for enzymes inactivated by PA Ao r cellulose.…”

mentioning

confidence: 99%

“…Ther eaction proceeds at 25 8 8Cu ntil dry and the BSA-Paper is ready for attachment of BSA and laccase in the second layer, which is then dried at 25 8 8Ca nd washed. [4] SEM (Scheme 1) shows the unchanged morphology of the coated fibers when compared to unmodified filter paper (Supporting Information, Figure S1). TheB SA in the second layer also functions as as pacer, so that Lacc-Lacc crosslinking is minimized, and forms ah ighly crosslinked network facilitating enzyme stabilization by retaining enzyme conformation under undesirable conditions.…”

mentioning

confidence: 99%

“…We previously reported that glucose oxidase (GOx) and peroxidase (HRP) can be interlocked in filter paper by covalent conjugation with poly(acrylic acid) (PAA) via carbodiimide chemistry without any cellulose functionalization. [4] This method however, is not suitable for enzymes with insufficient lysine residues to attach to the carboxylic acid groups of PA A, or for enzymes inactivated by PA Ao r cellulose.…”

mentioning

confidence: 99%

“…Theporosity of BSA-Paper was not significantly altered from that of uncoated paper,a st he total protein loaded is too small to influence the pore diameter. [4] SEM (Scheme 1) shows the unchanged morphology of the coated fibers when compared to unmodified filter paper (Supporting Information, Figure S1). Liquid media can therefore quickly and freely access the internal matrix networks.…”

mentioning

confidence: 99%

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A Modular Approach for Interlocking Enzymes in Whatman Paper

et al. 2018

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A potentially universal approach is presented for enzyme attachment to cellulose that significantly enhances enzyme stability while retaining high activity, and involves no chemical functionalization of cellulose. Bovine serum albumin (BSA) was interlocked in cellulose to form a protein-friendly surface (named BSA-Paper), while also providing COOH and NH groups for subsequent attachment of enzymes. The desired enzyme is then mixed with additional BSA and interlocked on BSA-Paper. The second BSA layer dilutes and crosslinks the enzyme for improved stability. Laccase was tested as a model enzyme for interlocking on BSA-Paper, and was found to retain over 100 % activity and was 240 times more stable at 25 °C (half life=180 d) than laccase. This new approach was also tested with a few other enzymes with encouraging results, thus providing a potentially universal method for stabilization of enzymes on cellulose with retention of high activities.

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Leveraging the Power of Enzymes in Engineered Dead and Living Materials

Shannon,

Zhou,

Perriman

2024

Adv Funct Materials

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Significant advances are being made in the incorporation of enzymes into functional materials, which leverage their inherent substrate specificity, efficient catalytic activity, and sustainable origin. These engineered “dead” materials, however, lack the incredible systems‐level control of enzyme activity that living organisms have evolved over millennia. This gap is now being bridged by the rapidly emerging field of engineered living materials (ELMs), which couples the tools of advanced synthetic biology with modern materials science. In this review, the impressive array of methodologies used to fabricate the extensive library of functional enzyme‐based dead materials is discussed, and the design strategies that facilitate their creation unpacked. The spectacular suite of natural and synthetic genetic and post‐translational control systems for enzymes in living organisms is then described. Finally, key recent examples of ELMs that utilize enzyme activity are reviewed, highlighting the central role of the living component in providing responsivity and adaptability to this new class of materials.

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Covalent interlocking of glucose oxidase and peroxidase in the voids of paper: enzyme–polymer “spider webs”

Cited by 32 publications

References 27 publications

Protein Biophosphors: Biodegradable, Multifunctional, Protein‐Based Hydrogel for White Emission, Sensing, and pH Detection

Protein Biophosphors: Biodegradable, Multifunctional, Protein‐Based Hydrogel for White Emission, Sensing, and pH Detection

A Modular Approach for Interlocking Enzymes in Whatman Paper

Leveraging the Power of Enzymes in Engineered Dead and Living Materials

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