Multi-enzyme systems: bringing enzymes together in vitro

Schoffelen, Sanne; Hest, Jan C. M. van

doi:10.1039/c1sm06452e

Cited by 239 publications

(202 citation statements)

References 112 publications

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“…The technical performance includes enzyme stability, substrate specificity, enantioselectivity, and reactivity (Mateo et al 2007;Schoffelen and van Hest 2012). The target of most immobilization practices is mainly to achieve fewer side reactions, high tolerance of structural variation of the substrates, high productivity and space-time yield, and high durability of the biocatalyst (Cao et al 2003).…”

Section: Immobilized and Co-immobilized Systemsmentioning

confidence: 99%

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Lignocellulases: a review of emerging and developing enzymes, systems, and practices

Obeng

Adam

Budiman

et al. 2017

Bioresour. Bioprocess.

132

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The highly acclaimed prospect of renewable lignocellulosic biocommodities as obvious replacement of their fossilbased counterparts is burgeoning within the last few years. However, the use of the abundant lignocellulosic biomass provided by nature to produce value-added products, especially bioethanol, still faces significant challenges. One of the crucial challenging factors is in association with the expression levels, stability, and cost-effectiveness of the cellulose-degrading enzymes (cellulases). Interestingly, several recommendable endeavors in the bid to curb these challenges are in pursuance. However, the existing body of literature has not well provided the updated roadmap of the advancement and key players spearheading the current success. Moreover, the description of enzyme systems and emerging paradigms with high prospects, for example, the cell-surface display system has been ill-captured in the literature. This review focuses on the lignocellulosic biocommodity pathway, with emphasis on cellulase and hemicellulase systems. The paradigm shift towards cell-surface display system and its emerging recommendable developments have also been discussed. The attempts in supplementing cellulase with other enzymes, accessory proteins, and chemical additives have also been discussed. Moreover, some of the prominent and influential discoveries in the cellulase fraternity have been discussed.

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Section: Immobilized and Co-immobilized Systemsmentioning

confidence: 99%

“…There are also reviews on cellulase engineering and other in vitro strategies towards improving the functionality of cellulases (Bayer et al 2008;Himmel et al 2010;Schoffelen and van Hest 2012). However, the fraternity still faces challenges in terms of robustness, hydrolysis efficiency, and cost of these crucial enzymes.…”

Section: Introductionmentioning

confidence: 99%

Lignocellulases: a review of emerging and developing enzymes, systems, and practices

Obeng

Adam

Budiman

et al. 2017

Bioresour. Bioprocess.

132

View full text Add to dashboard Cite

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“…With the latter, this disadvantage is obviated through the covalent linkage of the enzyme to the support but may result in significantly reduced activity, particularly if the covalent linkages are at a site adjacent to or at the active site of 35 the enzyme. Spatial resolution 4 of enzyme immobilization, where the enzyme is immobilised at specific locations on a surface, is in general limited to defining small areas such as the surfaces of nanoparticles which are then utilized as enzyme carriers. 2 Typically the entire surface area of the particle is utilized and 40 often multilayer adsorption can occur.…”

Section: Introductionmentioning

confidence: 99%

“…15 The encapsulation of enzymes in sol gels has been widely utilised 19 . Sol-gels are formed via hydrolysis and condensation of a precursor species such as Si(OC 2 H 5 ) 4 . Hydrolysis of Si-OC 2 H 5 in the presence of acid results in the formation of Si-O-Si species which can then undergo further reaction to from a three 20 dimensional silicate network.…”

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confidence: 99%

Immobilisation of enzymes on mesoporous silicate materials

2013

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Mesoproous silicates (MPS) are attractive materials for the immobilisation of enzymes. They possess 5 ordered pore structures, narrow pore size distributions, large surface areas, high stability and can be chemically modified with various functional groups. The properties of MPS materials are reviewed in terms of their ability to act as supports for enzymes for use in biocatalysis with a particular focus on the ability to tailor the surface functionalization of the MPS to suit a specific enzyme. While many reports of the immobilisation of enzymes on MPS have been described, their use as biocatalytic supports is limited. 10Large scale reactors based on MPS will require continuous flow systems where the properties of the support can be tailored while allowing fluid flow at reasonable low pressure.

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“…5,6 Such highly concerted mechanisms possess advantages including maintaining high local concentration of the intermediates and reducing diffusion losses during the long transportation, which are especially critical for highly unstable reactive intermediates. 4,7 To mimic this process in vitro, researchers have developed various strategies and approaches to spatially colocalize multiple enzymes on carriers to achieve enhancement in reaction kinetics along with the ability to direct the reaction pathway. 8−13 In one example, a multiple-enzyme extract adsorbed on chitin exhibited higher stability over a range of temperature and pH than did the soluble multienzyme extract.…”

Section: ■ Introductionmentioning

confidence: 99%

Block Copolymer-Quantum Dot Micelles for Multienzyme Colocalization

et al. 2012

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To mimic the structure and functionality of multienzyme complexes, which are widely present in Nature, Pluronic-based micelles were designed to colocalize multiple enzymes. To stabilize the micelles as well as to enable characterization of single enzyme immobilization and multienzyme colocalization by FÃ ¶rster resonance energy transfer (FRET), quantum dots (QDs) were incorporated into the micelles to form Pluronic-QD micelles using a novel microreactor. Model enzymes glucose oxidase (GOX) and horseradish peroxidase (HRP) were respectively labeled with fluorescent dyes. The results indicated that FRET occurred between the QDs and dyes that labeled each type of enzyme in single enzyme immobilization studies as well as between the dyes in colocalization studies. These observations were consistent with increases in micelle size after adsorption of dye-enzymes as verified by dynamic light scattering. In addition, the activity of single enzymes was retained after immobilization. An optimized colocalization process improved the overall conversion rate by approximately 100% compared to equivalent concentrations of free enzymes in solution. This study demonstrates a versatile platform for multienzyme colocalization and an effective strategy to characterize multienzyme immobilization and colocalization, which can be applicable to many other multienzyme systems. ABSTRACT: To mimic the structure and functionality of multienzyme complexes, which are widely present in Nature, Pluronic-based micelles were designed to colocalize multiple enzymes. To stabilize the micelles as well as to enable characterization of single enzyme immobilization and multienzyme colocalization by Forster resonance energy transfer (FRET), quantum dots (QDs) were incorporated into the micelles to form Pluronic-QD micelles using a novel microreactor. Model enzymes glucose oxidase (GOX) and horseradish peroxidase (HRP) were respectively labeled with fluorescent dyes. The results indicated that FRET occurred between the QDs and dyes that labeled each type of enzyme in single enzyme immobilization studies as well as between the dyes in colocalization studies. These observations were consistent with increases in micelle size after adsorption of dye-enzymes as verified by dynamic light scattering. In addition, the activity of single enzymes was retained after immobilization. An optimized colocalization process improved the overall conversion rate by approximately 100% compared to equivalent concentrations of free enzymes in solution. This study demonstrates a versatile platform for multienzyme colocalization and an effective strategy to characterize multienzyme immobilization and colocalization, which can be applicable to many other multienzyme systems.

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Multi-enzyme systems: bringing enzymes together in vitro

Cited by 239 publications

References 112 publications

Lignocellulases: a review of emerging and developing enzymes, systems, and practices

Lignocellulases: a review of emerging and developing enzymes, systems, and practices

Immobilisation of enzymes on mesoporous silicate materials

Block Copolymer-Quantum Dot Micelles for Multienzyme Colocalization

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