Production of self-immobilised enzyme microspheres using microfluidics

Mbanjwa, MB; Land, Kevin; Windvoel, Thobile; Papala, Prince M.; Fourie, Louis; Korvink, Jan G.; Visser, Daniel F.; Brady, Dean

doi:10.1016/j.procbio.2018.03.002

Cited by 9 publications

(7 citation statements)

References 39 publications

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“…In this study, the total conversion proportion of potato starch was relative low because the residual activity of CGTase was low and the barrier to potato starch permeating as potato starch was macromolecular. When the enzyme exits in the form of CLEA, the internal and external structures had been changed . Some structural changes in the enzyme introduced by the immobilization procedure, the mass transfer and diffusion become rate‐limiting, causing interaction to change between the immobilized enzyme and substance, as well as to the lower accessibility of the substrate to the active site of immobilized enzyme .…”

Section: Resultsmentioning

confidence: 99%

“…When the enzyme exits in the form of CLEA, the internal and external structures had been changed. 24 Some structural changes in the enzyme introduced by the immobilization procedure, the mass transfer and diffusion become rate-limiting, causing interaction to change between the immobilized enzyme and substance, as well as to the lower accessibility of the substrate to the active site of immobilized enzyme. 25 Considering the potato starch conversion proportion of CLEA-CGTase was lower than that of the same amount of free CGTase, we deduced that this result was attributed to a starch permeation barrier, which is a common issue with immobilization of enzyme including CLEA-CGTase for macromolecular substrates.…”

Section: Production Of CD By Clea-cgtasementioning

confidence: 99%

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Cross‐linked enzyme aggregates of recombinant cyclodextrin glycosyltransferase for high‐purity β‐cyclodextrin production

Zhang

Mao

2019

J of Chemical Tech & Biotech

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BACKGROUND High‐purity α‐, β‐, or γ‐cyclodextrin (CD) production using cyclodextrin glycosyltransferase (CGTase) as biocatalyst to cyclize starch is advantageous owing to its low cost of purification and product specificity. Several approaches have been investigated to enhance product specificity of CGTase, such as new CGTase isolation, CGTase gene (cgt) mutation, and chemical addition. RESULTS Recombinant CGTase was cross‐linked by glutaraldehyde to obtain cross‐linked enzyme aggregates of recombinant CGTase (CLEA‐CGTase) at 85 °C because free CGTase showed high product specificity at 85 °C. The CLEA‐CGTase was prepared under the conditions 75 U mL−1 CGTase with 0.1% (v/v) glutaraldehyde for 10 min at 85 °C after optimization first, and produced a high proportion of β‐CD from soluble starch at 50 °C in the conversion system. The conversion process was carried out with 10 to 200 U mL−1 CLEA‐CGTase, resulting in 100% β‐CD from soluble starch after 420 min conversion of the potato starch solution at 50 °C. And the proportion of β‐CD was still higher than 90% when 8000 U mL−1 CLEA‐CGTase was added. CONCLUSIONS CLEA technology maintained CGTase conformation of 85 °C, and produced a high proportion of β‐CD at 50 °C. This research showed the CLEA technology kept the conformation of enzyme for specific product production. © 2018 Society of Chemical Industry

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Section: Resultsmentioning

confidence: 99%

Section: Production Of CD By Clea-cgtasementioning

confidence: 99%

Cross‐linked enzyme aggregates of recombinant cyclodextrin glycosyltransferase for high‐purity β‐cyclodextrin production

Zhang

Mao

2019

J of Chemical Tech & Biotech

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“…This affords uniform enzyme microspheres, aptly named 'spherezymes', with a particle diameter of 50 mm and a size distribution within 3% and more controlled structural features. 25 When using polymers in industrial processes, they should preferably be in the form of spherical beads as uniform as possible in terms of particle size distribution. Generally, industrial processes use particles larger than 100 micron that can fit either batch reactors or column reactors.…”

Section: Size Matters: Activity and The Particle Size Dilemmamentioning

confidence: 99%

New frontiers in enzyme immobilisation: robust biocatalysts for a circular bio-based economy

Basso²,

2021

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“…In a novel refinement on the cross-linking theme, Brady and coworkers [126,127] prepared Spherezymes by adding cross-linking agents to a water-in-oil emulsion of an aqueous solution of P. fluorescens lipase. The spherezyme particles exhibited good activity in organic solvents.…”

Section: Advantages Limitations and Optimization Of Cleasmentioning

confidence: 99%

CLEAs, Combi-CLEAs and ‘Smart’ Magnetic CLEAs: Biocatalysis in a Bio-Based Economy

2019

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Biocatalysis has emerged in the last decade as a pre-eminent technology for enabling the envisaged transition to a more sustainable bio-based economy. For industrial viability it is essential that enzymes can be readily recovered and recycled by immobilization as solid, recyclable catalysts. One method to achieve this is via carrier-free immobilization as cross-linked enzyme aggregates (CLEAs). This methodology proved to be very effective with a broad selection of enzymes, in particular carbohydrate-converting enzymes. Methods for optimizing CLEA preparations by, for example, adding proteic feeders to promote cross-linking, and strategies for making the pores accessible for macromolecular substrates are critically reviewed and compared. Co-immobilization of two or more enzymes in combi-CLEAs enables the cost-effective use of multiple enzymes in biocatalytic cascade processes and the use of "smart" magnetic CLEAs to separate the immobilized enzyme from other solids has raised the CLEA technology to a new level of industrial and environmental relevance. Magnetic-CLEAs of polysaccharide-converting enzymes, for example, are eminently suitable for use in the conversion of first and second generation biomass.Catalysts 2019, 9, 261 2 of 31 and deprotection steps. This affords synthetic routes that, compared with conventional organic syntheses, are more step economic, more energy efficient, generate less waste and provide products in exquisite stereochemical purities that are difficult to compete with.Consequently, in the last two decades biocatalysis has emerged as an important technology for meeting the growing demand for green and sustainable processing [2][3][4]. This embraces the whole gamut of chemicals manufacture, from the enantioselective synthesis of chiral drugs [5][6][7][8][9][10][11][12] to the conversion of renewable biomass to liquid fuels and commodity chemicals [13][14][15]. This raises the perennial question: if biocatalysis is so superior why has it not been extensively used in the past? The answer is simple: thanks to advances in molecular biology and biotechnology, biocatalysis has undergone a spectacular leap forward in the last two decades:-Many more enzymes have been identified through (meta)genome mining, that is the in silico analysis of publicly accessible genome sequence data bases that have been generated as a result of next generation genome sequencing [16]. -Advances in gene synthesis have enabled the synthesis of identified genes, ready for cloning into a host production organism, in a few weeks at relatively low cost. This has significantly reduced the cost of development and subsequent production of enzymes at industrial scale. -Advances in protein engineering, using directed evolution techniques [17,18], have enabled optimization of enzyme performance under the challenging conditions encountered in industrial-scale processes, namely, high (stereo)selectivities, activities and space-time yields with non-natural substrates at high substrate concentrations, in the presence of organic solve...

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Production of self-immobilised enzyme microspheres using microfluidics

Cited by 9 publications

References 39 publications

Cross‐linked enzyme aggregates of recombinant cyclodextrin glycosyltransferase for high‐purity β‐cyclodextrin production

Cross‐linked enzyme aggregates of recombinant cyclodextrin glycosyltransferase for high‐purity β‐cyclodextrin production

New frontiers in enzyme immobilisation: robust biocatalysts for a circular bio-based economy

CLEAs, Combi-CLEAs and ‘Smart’ Magnetic CLEAs: Biocatalysis in a Bio-Based Economy

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