Immobilization of Enzymes on Hetero-Functional Supports: Physical Adsorption Plus Additional Covalent Immobilization

Trobo-Maseda, Lara; Orrego, Alejandro H.; Romero‐Fernández, María; Guisán, José M.; Rocha-Martín, Javier

doi:10.1007/978-1-0716-0215-7_10

Cited by 7 publications

(4 citation statements)

References 19 publications

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“…However, heterofunctional supports have significant advantages, and in a study by Trobo-Maseda et al, heterofunctional amino-epoxy and amino-glyoxyl groups were used. In first step, the enzyme is attached to the support, and in the second step, the intramolecular attachment between the enzyme and the support is enhanced [25]. Heterofunctional support generally elevates enzyme specificity and stability to demonstrate high-performance activity.…”

Section: Crosslinked Enzyme Aggregatesmentioning

confidence: 99%

Bio-Enzyme Hybrid with Nanomaterials: A Potential Cargo as Sustainable Biocatalyst

et al. 2023

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With advancements in bionanotechnology, the field of nanobiocatalysts has undergone rapid growth and revolutionized various nanomaterials as novel and fascinating nanocarriers for enzyme immobilization. Nanotubes, nanofibers, nanopores, nanoparticles, and nanocomposites have been successfully developed and used as nanocarriers. The construction of robust nanobiocatalysts by combining enzymes and nanocarriers using various enzyme immobilization techniques is gaining incredible attention because of their extraordinary catalytic performance, high stability, and ease of reusability under different physical and chemical conditions. Creating appropriate surface chemistry for nanomaterials promotes their downstream applications. This review discusses enzyme immobilization on nanocarriers and highlights the techniques, properties, preparations, and applications of nanoimmobilized enzymes.

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Section: Crosslinked Enzyme Aggregatesmentioning

confidence: 99%

Bio-Enzyme Hybrid with Nanomaterials: A Potential Cargo as Sustainable Biocatalyst

et al. 2023

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“…The protein solution was prepared in 5 mM phosphate buffer pH 7. After being washed, the immobilize enzyme was incubated with 1 ml of 5 mM NaHCO 3 buffer pH 9 for at least 2 h. Afterwards, the support was washed and left in 4 ml blocking buffer (prepared as previously described) for an overnight (Trobo-Maseda et al, 2020).…”

Section: Enzyme Immobilizationmentioning

confidence: 99%

Fusion of Glutamate Dehydrogenase and Formate Dehydrogenase Yields a Bifunctional Efficient Biocatalyst for the Continuous Removal of Ammonia

et al. 2021

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A novel fusion protein has been rationally designed, combining the hexameric glutamate dehydrogenase from Clostridium symbiosum with the dimeric formate dehydrogenase from Candida boidinii. The former enzyme consumes ammonia for the reductive amination of α-ketoglutarate using NADH, while the latter biocatalyst regenerates continuously the cofactor. This enzymes fusion opens new perspectives for the detection and the removal of ammonia. The bifunctional biocatalyst has been successfully created, expressed, and then characterized. The two fused protein domains retained identical properties and catalytic activity of the individual enzymes. Additionally, the immobilization on a methacrylate resin optimized the assembly providing a reusable and stable biocatalyst. This is an example of immobilization of a fusion protein, so that efficiency and sustainability of the process are enhanced. The immobilized biocatalyst could be recycled 10 times retaining still half of the initial activity. Such preparation outperforms the co-immobilized wild-type enzymes in the conversion of 300 mM of ammonia, which could be carried out also in continuous mode.

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“…These heterofunctional supports emerge as an excellent solution to coimmobilize multienzyme systems on the same surface where each enzyme is attached to the support through its optimal immobilization chemistry. The vast majority of heterofunctional supports offer the combination of only two reactive groups: one (i.e., ionic, hydrophobic, metal chelate groups) to drive the enzyme adsorption and the other (i.e., epoxy, aldehyde, glyoxyl, and vinyl groups) to react with the exposed nucleophilic residues on the enzyme surface to form covalent and irreversible bonds. , The combination of these two groups allows a two-step enzyme immobilization, in which the enzyme is first absorbed very quickly to the support (close contact) and then irreversible covalent attachment between the enzyme and the support is formed . Although heterofunctional supports have been mainly harnessed to accomplish the multivalent covalent immobilization of single enzymes at mild conditions, recent trends are more focused on their use as a chassis for the coimmobilization of multienzyme systems controlling their spatial organization. ,, …”

Section: Introductionmentioning

confidence: 99%

Multienzyme Coimmobilization on Triheterofunctional Supports

2023

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Immobilized multienzyme systems are gaining momentum in applied biocatalysis; however, the coimmobilization of several enzymes on one carrier is still challenging. In this work, we exploited a heterofunctional support activated with three different chemical functionalities to immobilize a wide variety of different enzymes. This support is based on agarose microbeads activated with aldehyde, amino, and cobalt chelate moieties that allow a fast and irreversible immobilization of enzymes, enhancing the thermostability of most of the heterogeneous biocatalysts (up to 21-fold higher than the soluble one). Furthermore, this trifunctional support serves to efficiently coimmobilize a multienzyme system composed of an alcohol dehydrogenase, a reduced nicotinamide adenine dinucleotide (NADH) oxidase, and a catalase. The confined multienzymatic system demonstrates higher performance than its free counterpart, achieving a total turnover number (TTN) of 1 × 105 during five batch consecutive cycles. We envision this solid material as a platform for coimmobilizing multienzyme systems with enhanced properties to catalyze stepwise biotransformations.

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Immobilization of Enzymes on Hetero-Functional Supports: Physical Adsorption Plus Additional Covalent Immobilization

Cited by 7 publications

References 19 publications

Bio-Enzyme Hybrid with Nanomaterials: A Potential Cargo as Sustainable Biocatalyst

Bio-Enzyme Hybrid with Nanomaterials: A Potential Cargo as Sustainable Biocatalyst

Fusion of Glutamate Dehydrogenase and Formate Dehydrogenase Yields a Bifunctional Efficient Biocatalyst for the Continuous Removal of Ammonia

Multienzyme Coimmobilization on Triheterofunctional Supports

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