Review of: Fred Höfler

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No planned obsolescence here. Hope, sweet hope. Furthermore, performing the reaction in H 2 18 O-enriched buffer resulted in the formation of 18 O-labeled R phenyl ethanol Figure 2 b,c.

This confirms that indeed the reaction medium serves as a source of H 2 O 2 and that reduction of O 2 from ambient air played a minor role in the H 2 O 2 formation.

R phenyl ethanol was the sole product observed, indicating that the selectivity of the biocatalyst was not impaired under the reaction conditions, particularly by the radioactivity.

A control reaction with R phenyl ethanol only under the irradiation showed that radiation-induced further oxidation of the primary enzyme product R phenyl ethanol to acetophenone can be ruled out.

Next, we further investigated some factors influencing the efficiency and robustness of the overall reaction Figure 3.

Increasing the biocatalyst concentration increased the product formation within the first hour Figure 3 a. This increase, however, was not linear and converged to approx.

Interestingly, this product formation rate was approx. This observation can be attributed to the irreversible peroxygenase step removing H 2 O 2 from the steady-state equilibrium.

These experiments, however, also revealed a poor long-term stability of the enzyme under the reaction conditions. Already after 1 h of reaction approx.

This assumption is supported by a considerable decrease in the characteristic Soret peak at nm indicative for an intact heme moiety Figure S3.

Analysis of an inactivated enzyme sample by native gel electrophoresis gave no indication for loss of the quaternary structure Figure S4. We reasoned that hydroxyl radicals formed during the physicochemical phase of the water radiolysis reaction, for example, the hydroxyl radicals, may oxidatively inactivate the catalytic heme functionality and thereby the biocatalyst.

Of course, also H 2 O 2 -dependent inactivation of the prosthetic group can also contribute to the observed inactivation. However, also the highly H 2 O 2 -stable, 35 V-dependent chloroperoxidase from Curvularia inaequalis Ci VCPO 36 was inactivated under these conditions vide infra.

Suspecting intermediate radical species Figure 1 a as major contributors to the observed biocatalyst inactivation, we tested a range of different radical scavengers Figure 3 b.

Among these radical scavengers especially methanol, acrylamide, and formate enabled significantly increased product formation Figure 3 b.

The effect depended on the concentration of the radical scavenger as exemplified with methanol and formate Figures S5 and S6. We therefore also compared the time courses of the radioenzymatic reactions in the absence and presence of the radical scavengers methanol and formate Figure 3 c.

Most strikingly, the conversion of ethyl benzene to R phenyl ethanol was increased from approx. In the latter case, a turnover number for the biocatalyst of The dose rate of the radiation source directly influenced the product formation of the radioenzymatic reaction system Table 1.

The final product concentration and directly related to this also the turnover number of the enzyme directly correlated with the dose rate of the radioactivity source applied.

This may be due to a decreased radiolytic H 2 O 2 decomposition at lower dose rates, whereas the biocatalyst concentration remained constant.

Further experiments will be necessary to fully rationalize this observation. Pleasantly, the reactions performed with spent fuel element U also showed good robustness.

Table 1. Radioenzymatic Hydroxylation of Ethyl Benzene Using Different Radiation Sources a. Finally, we initially explored the substrate scope of the proposed radioenzymatic reaction scheme Figure 4.

For this, some further oxyfunctionalization reactions reported for r Aae UPO, such as epoxidation 37 as well as aliphatic 38 and aromatic hydroxylation reactions, were chosen.

Possibly, the radicals present in the reaction mixture lead to racemization of the epoxide product, but further investigations will be necessary to confirm this.

Again, a beneficial effect of formate on the product formation was observed Figure 4. To address the question whether too high H 2 O 2 concentrations may contribute to the abovementioned inactivation of r Aae UPO, we extended the enzyme scope of the proposed radioenzymatic reaction to the vanadium-dependent chloroperoxidase from C.

As H 2 O 2 as a cause for this can be ruled out, we assign this observation to Ci VCPO inactivation by hydroxyl radicals Figure S9.

In conclusion, we have demonstrated that radiolytic water splitting can be used to promote biocatalytic oxyfunctionalization reactions. The dose-rate-dependent steady-state concentration appears ideal to provide heme-dependent peroxygenases with suitable concentrations of H 2 O 2 that enable the reaction while minimizing the oxidative inactivation.

This advantage, at least in the present setup, is compensated by the radical-induced inactivation of the biocatalyst, this is also reflected by the comparably poor performance of the present system compared to other in situ H 2 O 2 generation systems Table S1.

Compared to other radical-generating H 2 O 2 generation systems, the turnover numbers observed here compare very well.

The radical inactivation of the biocatalysts represents an apparent shortcoming of the current setup. In future experiments, we will address this by physical separation of the biocatalyst from the radiation source.

Flow chemistry appears a particularly attractive technical solution. Although this approach at first sight may appear as a lab curiosity, we believe that it may actually bear some practical relevance.

In this study, we have demonstrated that spent fuel elements can drive peroxygenase-catalyzed reactions. Considering the annually increasing amounts of radioactive waste and its persistence, the proposed radioenzymatic approach may represent a possibility to productively utilize nuclear waste.

Furthermore, it should be kept in mind that globally a variety of different radiation sources are used commercially. For instance, 60 Co units are used for sterilization and electron beams for various applications and research nuclear reactors more than worldwide.

Experimental Section. ARTICLE SECTIONS Jump To. The evolved, unspecific peroxygenase from Agrocybe aegrita r Aae UPO was obtained from fermentation of recombinant Pichia pastoris as previously described.

The vanadium-dependent chloroperoxidase from C. All radiochemical experiments were performed by placing 2 mL GC vials filled with 1 mL of the reaction mixture next to the radioactivity source Figure S At intervals, samples were removed from the radiation source and analyzed.

Supporting Information. Author Information. Huanhuan Liu - Radiation Science and Technology, Delft University of Technology, Mekelweg 15, JB Delft, The Netherlands.

Morten M. Antonia G. Denkova - Radiation Science and Technology, Delft University of Technology, Mekelweg 15, JB Delft, The Netherlands.

Water Radiolysis: Influence of Oxide Surfaces on H 2 Production under Ionizing Radiation. Water Basel, Switzerland , 3 , CODEN: WATEGH ; ISSN: MDPI AG.

A review. The radiolysis of water due to ionizing radiation results in the prodn. A brief history of the development of the understanding of water radiolysis is presented, with a focus on the H2 prodn.

This H2 prodn. Different parameters accounting for this behavior are presented. Polymer-gel formation and reformation on irradiation of tertiary-butyl acrylate.

Radiation Physics and Chemistry , 97 , CODEN: RPCHDM ; ISSN: X. Elsevier B. The purpose of the present research was to provide a radiation-chem.

The radiation-induced polymn. The fractional monomer-to-polymer conversion, CM, detd. After removal of monomer by evacuation, the gel can be reformed by adding a vol.

The dose and dose rate dependence of radiation-induced monomer conversion in the reformed gel show no evidence of a discontinuity caused by the intervening evacuation and reformation procedures.

Polymerization induced by ionizing radiation at low temperatures. Evidence for the simultaneous existence of ionic and free-radical mechanisms in the polymerization of styrene and 2,4-dimethylstyrene.

Journal of Physical Chemistry C , 34 , CODEN: JPCCCK ; ISSN: American Chemical Society. Dihydrogen H2 prodn. Four types of zeolites were comparatively examd.

High yields of H2 were obsd. The difference in the H2 yields was considerable at wt. The exptl. The H2 yields were compared with chem.

On the other hand, under aerated conditions, the H2 yields from the zeolites were lower than under anoxic conditions probably due to the H2O2 that was produced by water radiolysis.

Moreover, the difference between the zeolites in H2 yields was suppressed at water fractions above 50 wt. The comparable H2 yields suggested yet another reaction pathway for H2 prodn.

Green oxidation with aqueous hydrogen peroxide. Chemical Communications Cambridge, United Kingdom , 16 , CODEN: CHCOFS ; ISSN: Royal Society of Chemistry.

H2O2 is an ideal oxidant, when coupled with a tungstate complex and a quaternary ammonium hydrogensulfate as an acidic phase-transfer catalyst. It oxidizes alcs.

Peroxygenases en route to becoming dream catalysts. What are the opportunities and challenges? Current Opinion in Chemical Biology , 37 , CODEN: COCBF4 ; ISSN: Peroxygenases are promising catalysts for preparative oxyfunctionalization chem.

Though many interesting applications have been reported, today 'we have only scratched the surface' and significant efforts are necessary to solve issues related to selectivity of the wild type enzymes and low product titers.

For this, further elucidation of the vast natural diversity as well as protein and reaction engineering approaches are discussed. Oxidations catalyzed by fungal peroxygenases.

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