Enhancing promiscuous chemistries of a Schiff-base forming enzyme by divergent evolution

Macdonald, Duncan Stuart; Hilvert, Donald

doi:10.1016/bs.mie.2020.06.014

Cited by 2 publications

(2 citation statements)

References 38 publications

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“…Due to their mild reaction conditions and availability, they are often considered more valuable than classical catalysts. To justify the balance of enzymatic potential and the hazardous nature of hydrogen peroxide we decide to adopt the previously described technique [14] for the rapid determination of the perhydrolytic activity of an enzyme.…”

Section: Resultsmentioning

confidence: 99%

“…Uncovering the new role of enzyme which is different from their natural properties is highly active, [14] but the development of fast and simple screening assay for enzymatic divergent is still demanding. In particular, spectrophotometric methods are well suited to the enzymatic assay, because they are sensitive and allow to avoid the background signals.…”

Section: Introductionmentioning

confidence: 99%

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Efficient Assay for the Detection of Hydrogen Peroxide by Estimating Enzyme Promiscuous Activity in the Perhydrolysis Reaction

Wilk

Ostaszewski

2021

ChemBioChem

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Hydrogen peroxide is an ideal oxidant in view of its availability, atom economy, or green aspects. Furthermore, it is produced by the cell mitochondria and plays a meaningful role in controlling physiological processes, but its unregulated production leads to the destruction of organs. Due to its diverse roles, a fast and selective method for hydrogen peroxide detection is the major limitation to preventing the negative effects caused by its excess. Therefore, we aimed to develop an efficient assay for the detection of H 2 O 2 . For this purpose, we combined the enzymatic method for the detection of hydrogen peroxide with the estimation of the promiscuity of various enzymes. We estimated the activity of an enzyme in the reaction of pnitrophenyl esters with hydrogen peroxide resulting in the formation of peracid. To our knowledge, there is no example of a simple, multi-sensor demonstrating the promiscuous activity of an enzyme and detecting hydrogen peroxide in aqueous media.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Efficient Assay for the Detection of Hydrogen Peroxide by Estimating Enzyme Promiscuous Activity in the Perhydrolysis Reaction

Wilk

Ostaszewski

2021

ChemBioChem

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Witnessing the Birth of Directed Evolution of Stereoselective Enzymes as Catalysts in Organic Chemistry

Reetz

2022

Adv Synth Catal

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This invited essay outlines how the idea of directed evolution of stereoselective enzymes was born and implemented experimentally at the Max-Planck-Institut für Kohlenforschung in Mülheim/ Germany during the period 1994-1998, a time when Andreas Pfaltz was present in the Institute. As the new and sole director of the MPI, I initiated new research projects, and also started to restructure the Institute with the establishment of five departments, all dedicated to some form of catalysis and to be led by five independent directors. Andreas Pfaltz was the first director whom I hired, heading the Department of Homogeneous Catalysis. During his stay in Mülheim until 1998, the Pfaltz group invented effective chiral ligands for a number of particularly challenging enantioselective transformations. During this period, Andreas Pfaltz witnessed the birth and development of directed evolution of stereoselective enzymes as a new research direction in the Reetz group. Indeed, he was one of the very few organic chemists who realized at the time that a door to a novel research area had been opened. The widespread application of enzymes was hampered for decades due to limited enantio-, diastereo-, and regioselectivity, which was the reason why organic chemists were not interested in biocatalysis. This attitude slowly changed with the advent of directed evolution of stereoselective enzymes in 1997, in a publication from the Reetz group. Methodology development with emphasis on stereo-and regioselectivity as well as activity followed, the techniques and strategies becoming more and more rational. Today, semirational approaches and so-called rational enzyme design have merged, as evidenced, inter alia, by the development of focused rational iterative site-specific mutagenesis (FRISM). The toolbox of organic chemists now includes enzymes, primarily because the possibility of controlling stereoselectivity by protein engineering has ensured reliability when facing synthetic challenges. 1. Introductory Remarks 2. The Birth of Directed Evolution of Stereoselective Enzymes 3. The Crucial Role of Methodology Development in Directed Evolution of Selective Enzymes as Catalysts in Organic Chemistry 4. Final Message

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Enhancing promiscuous chemistries of a Schiff-base forming enzyme by divergent evolution

Cited by 2 publications

References 38 publications

Efficient Assay for the Detection of Hydrogen Peroxide by Estimating Enzyme Promiscuous Activity in the Perhydrolysis Reaction

Efficient Assay for the Detection of Hydrogen Peroxide by Estimating Enzyme Promiscuous Activity in the Perhydrolysis Reaction

Witnessing the Birth of Directed Evolution of Stereoselective Enzymes as Catalysts in Organic Chemistry

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