Synergistic catalysis in an artificial enzyme by simultaneous action of two abiological catalytic sites

Zhou, Z. Hong; Roelfes, Gérard

doi:10.1038/s41929-019-0420-6

Cited by 146 publications

(141 citation statements)

References 38 publications

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“…Another emerging future trend is the development of enzymes with genetically encoded unnatural amino acids (reviewed by Burgener et al, 2020;Drienovská and Roelfes, 2020 and also see Yu et al, 2018) and put into practice by Zhou and Roelfes (2020) who inserted a non-natural amino acid at the catalytic site of the transcription factor LmrR and with a complex containing copper, was able to activate reaction partners in the Michael addition reaction which creates carbon-carbon bonds. These experiments are single enzyme reactions at present but there is discussion on their incorporation into enzyme cascades which will probably include CFS.…”

Section: Discussionmentioning

confidence: 99%

Cell-Free Biocatalysis for the Production of Platform Chemicals

2020

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Genetically engineered host bacteria have an extensive history for the production of specific proteins including the synthesis of single enzymes for the modification of compounds produced for industrial purposes by biological or chemical processes. Such processes have been developed largely through the process of discovery. The ability to assemble multiple enzymes into synthetic pathways is a new development aided by the synthetic biology approach of constructing and assembling suitable enzymes into pathways that may or may not occur in Nature to provide a highimpact platform for bio-manufacturing of chemicals, biofuels and pharmaceuticals. Industry has depended on chemical catalysts because of the known constraints experienced frequently with biological catalysts but when high stereochemistry, mild synthesis conditions and environmentally friendly processes are significant, application of enzymes is preferred, especially in the case of drug synthesis where enantioselectivity is important. However, many whole cell production processes are beset by toxicity problems, metabolite competition, the production of side products, sub-optimal enzyme ratios and varying temperature optima. As a result, a cell-free biocatalysis allows the manipulation of substrate ratios, provision of regenerated cofactors and adjustment of high energy flux ratios that are difficult or impossible to control in whole cell synthesis. We discuss here the construction of cell free biocatalytic pathways as added free enzymes or multi-enzyme modules that may contain heterologous catalysts. We examine the status of applications leading to commercialisation, emphasizing the importance of economical cofactor regeneration. Nevertheless, problems remain, particularly protein post-translational modifications including glycosylation, phosphorylation, ubiquitination, acetylation, proteolysis, etc. The National Renewable Energy Laboratory and Pacific North West Laboratory have published lists of top value-added chemicals from biomass that include glucaric acid. There have been few reports on the combination of synthetic biology and cell-free protein synthesis to set up pathways for these valuable intermediate compounds. This observation is despite the existence of at least one large scale but specialized cell-free production of antibody conjugates. This review will provide a description of one successful attempt at the cell-free production of glucaric acid and will evaluate progress for other key intermediate and platform chemicals.

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Section: Discussionmentioning

confidence: 99%

Cell-Free Biocatalysis for the Production of Platform Chemicals

2020

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“…This can be exemplified by the recent development of LmrR modified with dual catalytic groups. 122 Genetic code expansion can also be used to introduce novel catalytic functionalities into a de novo designed enzyme active site. This has been demonstrated in a recently reported artificial metalloenzyme, in which the designed active site includes the unnatural amino acid bipyridylalanine for metal binding.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, the p -aminophenylalanine/LmrR system has been further modified for a novel dual substrate activation strategy. 122 Through combination with a supramolecularly bound Lewis acidic Cu( ii ) complex, the resulting artificial enzyme was able to mediate a Michael reaction that involves both formation of a Cu-enolate and an organocatalytic iminium intermediate. Yields of this novel reaction mode were up to 90%, with d.r.…”

Section: Genetic Code Expansionmentioning

confidence: 99%

Enabling protein-hosted organocatalytic transformations

et al. 2020

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“…Combination of the beneficial mutations finally achieved the LmrR_pAF_RMHL variant, in which four additional mutations were incorporated into LmrR(V15pAF), displaying a k cat value 91-times higher than that of its parent. Afterwards, an ArM based on LmrR(V15pAF) was developed, as described later in this review [ 94 ].…”

Section: Recent Progress On the Various Reactions Catalyzed By Armmentioning

confidence: 99%

Artificial Metalloenzymes: From Selective Chemical Transformations to Biochemical Applications

Himiyama

Okamoto

2020

Molecules

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Artificial metalloenzymes (ArMs) comprise a synthetic metal complex in a protein scaffold. ArMs display performances combining those of both homogeneous catalysts and biocatalysts. Specifically, ArMs selectively catalyze non-natural reactions and reactions inspired by nature in water under mild conditions. In the past few years, the construction of ArMs that possess a genetically incorporated unnatural amino acid and the directed evolution of ArMs have become of great interest in the field. Additionally, biochemical applications of ArMs have steadily increased, owing to the fact that compartmentalization within a protein scaffold allows the synthetic metal complex to remain functional in a sea of inactivating biomolecules. In this review, we present updates on: (1) the newly reported ArMs, according to their type of reaction, and (2) the unique biochemical applications of ArMs, including chemoenzymatic cascades and intracellular/in vivo catalysis. We believe that ArMs have great potential as catalysts for organic synthesis and as chemical biology tools for pharmaceutical applications.

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Synergistic catalysis in an artificial enzyme by simultaneous action of two abiological catalytic sites

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Cited by 146 publications

References 38 publications

Cell-Free Biocatalysis for the Production of Platform Chemicals

Cell-Free Biocatalysis for the Production of Platform Chemicals

Enabling protein-hosted organocatalytic transformations

Artificial Metalloenzymes: From Selective Chemical Transformations to Biochemical Applications

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