A generic platform for the immobilisation of engineered biocatalysts

Thompson, Matthew P.; Derrington, Sasha R.; Heath, Rachel S.; Porter, Joanne L.; Mangas‐Sánchez, Juan; Devine, Paul N.; Truppo, Matthew D.; Turner, Nicholas J.

doi:10.1016/j.tet.2018.12.004

Cited by 81 publications

(102 citation statements)

References 38 publications

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“…In the preceding decade the efficacy and cost-effectiveness of the cross-linked enzyme aggregates (CLEA) methodology in the immobilization of a wide variety of enzymes, thereby enabling many applications of biocatalysis, has been amply demonstrated. With regard to the eight requirements that Turner and coworkers [40] recently outlined for the commercial viability of an immobilized enzyme in the pharmaceutical industry, CLEAs exhibit (i) high enzyme loading (>10 wt%); (ii) high activity recovery (>50%); (iii) no leaching under reaction conditions; (iv) good tolerance to organic solvents; (v) are eminently recyclable (>20 cycles); and (vi) result in a lower cost of good compared with alternatives. Mass transport of substrates is sometimes an issue, especially with macromolecular substrates, but techniques have been developed to alleviate this problem.…”

Section: Discussionmentioning

confidence: 99%

“…Turner and coworkers [40] recently outlined the requirements that an immobilized enzyme should meet for commercial viability of a process in the pharmaceutical industry: (i) high enzyme loading (>10 wt%), (ii) high activity recovery (>50%), (iii) no leaching under reaction conditions, (iv) tolerance to organic solvents, (v) recyclable (>20 cycles), (vi) good substrate mass transport and (vii) mechanically stable in batch and flow reactors and (viii) last but not least, it must result in a lower cost of goods compared with alternatives. The authors also pointed out that a method should have broad applicability over numerous enzyme classes and, as an absolute minimum, generality within the same enzyme class.…”

Section: Enzyme Immobilizationmentioning

confidence: 99%

“…Interestingly, the production of both immobilized enzymes involves a cross-linking step with glutaraldehyde. The latter is generally the cross-linker of choice as it is inexpensive, readily available and its use in cross-linking of proteins has GRAS (generally regarded as safe) status for use in, among other things, food and beverages processing [39].Turner and coworkers [40] recently outlined the requirements that an immobilized enzyme should meet for commercial viability of a process in the pharmaceutical industry: (i) high enzyme loading (>10 wt%), (ii) high activity recovery (>50%), (iii) no leaching under reaction conditions, (iv) tolerance to organic solvents, (v) recyclable (>20 cycles), (vi) good substrate mass transport and (vii) mechanically stable in batch and flow reactors and (viii) last but not least, it must result in a lower cost of goods compared with alternatives. The authors also pointed out that a method should have broad applicability over numerous enzyme classes and, as an absolute minimum, generality within the same enzyme class.…”

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confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Enzyme Immobilizationmentioning

confidence: 99%

mentioning

confidence: 99%

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

2019

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“…The rapidly increasing use of enzymes in industrial organic synthesis has led to a growing demand for immobilized enzymes. Turner and co‐workers recently delineated the requirements for the viability of a process using an immobilized biocatalyst in the pharmaceutical industry:…”

Section: Expanding the Scope And Potential Of Biocatalysismentioning

confidence: 99%

“…The rapidlyi ncreasing use of enzymes in industrial organic synthesis has led to ag rowing demandf or immobilized enzymes. Turner and co-workers [66] recently delineated the requirements for the viabilityo faprocess using an immobilized biocatalyst in the pharmaceutical industry: Immobilization of isolatede nzymes usually involves binding to ap refabricated carrier (support), such as as ynthetic polymer,b iopolymer,o ri norganic solid. [67] Owing to the introduction of large amounts of noncatalytic material, the use of ac arrier inevitably leads to "dilution" of activity.C onsequently, there is increasing interesti nc arrier-free immobilized enzymes, [68] that couple high catalyst productivities and spacetime yields with high stabilities and low production costs, because there are no additional costs of an (expensive) carrier.…”

Section: Betterformulation and Recycling Of Enzymes Through Immobilizmentioning

confidence: 99%

Broadening the Scope of Biocatalysis in Sustainable Organic Synthesis

2019

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This Review is aimed at synthetic organic chemists who may be familiar with organometallic catalysis but have no experience with biocatalysis, and seeks to provide an answer to the perennial question: if it is so attractive, why wasn't it extensively used in the past? The development of biocatalysis in industrial organic synthesis is traced from the middle of the last century. Advances in molecular biology in the last two decades, in particular genome sequencing, gene synthesis and directed evolution of proteins, have enabled remarkable improvements in scope and substantially reduced biocatalyst development times and cost contributions. Additionally, improvements in biocatalyst recovery and reuse have been facilitated by developments in enzyme immobilization technologies. Biocatalysis has become eminently competitive with chemocatalysis and the biocatalytic production of important pharmaceutical intermediates, such as enantiopure alcohols and amines, has become mainstream organic synthesis. The synthetic space of biocatalysis has significantly expanded and is currently being extended even further to include new‐to‐nature biocatalytic reactions.

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