General Overview on Immobilization Techniques of Enzymes for Biocatalysis

Romero‐Fernández, María; Paradisi, Francesca

doi:10.1002/9783527817290.ch12

Cited by 22 publications

(26 citation statements)

References 92 publications

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“…To this purpose, enzyme immobilization is an extremely interesting and promising technique recently employed in biotechnological industries, where enzymes are used for different purposes. It consists of the binding of free enzymes on different types of inert supports, the aim being to improve their stability and recyclability [42][43][44][45]. The use of immobilized enzymes creates practical, economic, and ecological advantages.…”

Section: Introductionmentioning

confidence: 99%

Enhanced Stability of Long-Living Immobilized Recombinant β-d-N-Acetyl-Hexosaminidase A on Polylactic Acid (PLA) Films for Potential Biomedical Applications

Calzoni

Cesaretti

Montegiove

et al. 2021

JFB

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β-d-N-acetyl-hexosaminidase (Hex, EC 3.2.1.52) is an acid hydrolase that catalyzes the cleavage of the β-1,4 bond in N-acetyl-d-galactosamine (Gal-NAc) and N-acetyl-d-glucosamine (Glc-NAc) from the non-reducing end of oligosaccharides and glycoconjugates. It is widely expressed in both the prokaryotic and eukaryotic world, where it performs multiple and important functions. Hex has antifungal activity in plants, is capable of degrading many biological substrates, and can play an important role in the biomedical field for the treatment of Tay-Sachs and Sandhoff diseases. With the aim being able to obtain a device with a stable enzyme, a method of covalent immobilization on polylactic acid (PLA) films was developed for the A isoform of the β-d-N-acetyl-hexosaminidase enzyme (HexA), produced in a recombinant way from Human Embryonic Kidney-293 (HEK-293) cells and suitably purified. An in-depth biochemical characterization of the immobilized enzyme was carried out, evaluating the optimal temperature, thermal stability, pH parameters, and Km value. Moreover, the stability of the enzymatic activity over time was assessed. The results obtained showed an improvement in terms of kinetic parameters and stability to heat for the enzyme following immobilization and the presence of HexA in two distinct immobilized forms, with an unexpected ability for one of them to maintain its functionality for a long period of time (over a year). The stability and functionality of the enzyme in its immobilized form are therefore extremely promising for potential biotechnological and biomedical applications.

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Section: Introductionmentioning

confidence: 99%

Enhanced Stability of Long-Living Immobilized Recombinant β-d-N-Acetyl-Hexosaminidase A on Polylactic Acid (PLA) Films for Potential Biomedical Applications

Calzoni

Cesaretti

Montegiove

et al. 2021

JFB

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“…However, mass transfer to and from the enzyme is often impaired. [131] Furthermore, leaching of the enzyme can occur. [132] Adsorption onto a solid support is another simple immobilization strategy.…”

Section: Enzyme Immobilization and Flow Chemistrymentioning

confidence: 99%

“…In those cases, coating the biocatalysts either before or after immobilization with other polymers, such as polyethylenimine (PEI) or activated dextran can help maintain the quaternary structure. [131] However, as with any immobilization, excessive rigidification of the enzyme can result in a loss of catalytic efficiency if structural rearrangements necessary for catalysis are impeded. In addition, the increased complexity of more sophisticated immobilization techniques often outweighs any benefits bestowed on the catalyst.…”

Section: Accepted Manuscriptmentioning

confidence: 99%

Looking Back: A Short History of the Discovery of Enzymes and How They Became Powerful Chemical Tools

Heckmann

Paradisi

2020

ChemCatChem

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Enzymatic approaches to challenges in chemical synthesis are increasingly popular and very attractive to industry given their green nature and high efficiency compared to traditional methods. In this historical review we highlight the developments across several fields that were necessary to create the modern field of biocatalysis, with enzyme engineering and directed evolution at its core. We exemplify the modular, incremental, and highly unpredictable nature of scientific discovery, driven by curiosity, and showcase the resulting examples of cutting-edge enzymatic applications in industry.

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“…Enzyme immobilization can be divided into three main categories: entrapment/encapsulation, cross-linking and binding to a support (Figure 4). Entrapment and encapsulation are low-cost techniques (mainly due to the use of polymers) in which enzymes are either trapped or retained inside a capsule, separated from the reaction medium [65]. Cross-linking enzyme (CLE) technology is a simple carrier-free technique performed with the addition of a cross-linking agent to an enzyme preparation, usually producing a rigid three-dimensional complex of enzymes [66].…”

Section: Immobilization Fundamentalsmentioning

confidence: 99%

“…Cross-linking enzyme (CLE) technology is a simple carrier-free technique performed with the addition of a cross-linking agent to an enzyme preparation, usually producing a rigid three-dimensional complex of enzymes [66]. Finally, binding to a support is based on the simple attachment of the enzyme to a solid support mediated by different possible chemical interactions (either covalent or non-covalent) [65]. In the particular case of cellulases, immobilization onto a support is one of the most interesting techniques since some strategic advantages can be gained after immobilization, which will be featured below.…”

Section: Immobilization Fundamentalsmentioning

confidence: 99%

Strategies towards Reduction of Cellulases Consumption: Debottlenecking the Economics of Lignocellulosics Valorization Processes

et al. 2021

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Lignocellulosic residues have been receiving growing interest as a promising source of polysaccharides, which can be converted into a variety of compounds, ranging from biofuels to bioplastics. Most of these can replace equivalent products traditionally originated from petroleum, hence representing an important environmental advantage. Lignocellulosic materials are theoretically unlimited, cheaper and may not compete with food crops. However, the conversion of these materials to simpler sugars usually requires cellulolytic enzymes. Being still associated with a high cost of production, cellulases are commonly considered as one of the main obstacles in the economic valorization of lignocellulosics. This work provides a brief overview of some of the most studied strategies that can allow an important reduction of cellulases consumption, hence improving the economy of lignocellulosics conversion. Cellulases recycling is initially discussed regarding the main processes to recover active enzymes and the most important factors that may affect enzyme recyclability. Similarly, the potential of enzyme immobilization is analyzed with a special focus on the contributions that some elements of the process can offer for prolonged times of operation and improved enzyme stability and robustness. Finally, the emergent concept of consolidated bioprocessing (CBP) is also described in the particular context of a potential reduction of cellulases consumption.

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General Overview on Immobilization Techniques of Enzymes for Biocatalysis

Cited by 22 publications

References 92 publications

Enhanced Stability of Long-Living Immobilized Recombinant β-d-N-Acetyl-Hexosaminidase A on Polylactic Acid (PLA) Films for Potential Biomedical Applications

Enhanced Stability of Long-Living Immobilized Recombinant β-d-N-Acetyl-Hexosaminidase A on Polylactic Acid (PLA) Films for Potential Biomedical Applications

Looking Back: A Short History of the Discovery of Enzymes and How They Became Powerful Chemical Tools

Strategies towards Reduction of Cellulases Consumption: Debottlenecking the Economics of Lignocellulosics Valorization Processes

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