Hydrogenase-based oxidative biocatalysis without oxygen.

Biocatalysis-based synthesis can provide a sustainable and clean platform for producing chemicals. Many oxidative biocatalytic routes require the cofactor NAD+ as an electron acceptor. To date, NADH oxidase (NOX) remains the most widely applied system for NAD+ regeneration. However, its dependence on O2 implies various technical challenges in terms of O2 supply, solubility, and mass transfer. Here, we present the suitability of a NAD+ regeneration system in vitro based on H2 evolution. The efficiency of the hydrogenase-based system is demonstrated by integrating it into a multi-enzymatic cascade to produce ketoacids from sugars. The total NAD+ recycled using the hydrogenase system outperforms NOX in all different setups reaching up to 44,000 mol per mol enzyme. This system proves to be scalable and superior to NOX in terms of technical simplicity, flexibility, and total output. Furthermore, the system produces only green H2 as a by-product even in the presence of O2.

Modeling-Assisted Design of Thermostable Benzaldehyde Lyases from Rhodococcus erythropolis for Continuous Production of α-Hydroxy Ketones.

Enantiopure α-hydroxy ketones are important building blocks of active pharmaceutical ingredients (APIs), which can be produced by thiamine-diphosphate-dependent lyases, such as benzaldehyde lyase. Here we report the discovery of a novel thermostable benzaldehyde lyase from Rhodococcus erythropolis R138 (ReBAL). While the overall sequence identity to the only experimentally confirmed benzaldehyde lyase from Pseudomonas fluorescens Biovar I (PfBAL) was only 65 %, comparison of a structural model of ReBAL with the crystal structure of PfBAL revealed only four divergent amino acids in the substrate binding cavity. Based on rational design, we generated two ReBAL variants, which were characterized along with the wild-type enzyme in terms of their substrate spectrum, thermostability and biocatalytic performance in the presence of different co-solvents. We found that the new enzyme variants have a significantly higher thermostability (up to 22 °C increase in T50 ) and a different co-solvent-dependent activity. Using the most stable variant immobilized in packed-bed reactors via the SpyCatcher/SpyTag system, (R)-benzoin was synthesized from benzaldehyde over a period of seven days with a stable space-time-yield of 9.3 mmol ⋅ L-1 ⋅ d-1 . Our work expands the important class of benzaldehyde lyases and therefore contributes to the development of continuous biocatalytic processes for the production of α-hydroxy ketones and APIs.

Erratum: Ten Bosch et al. Cold Atmospheric Pressure Plasma Comb-A Physical Approach for Pediculosis Treatment.

Due to an error during production and a corrupted data set, subsection 3.3.1 in the result section of the published paper [1] was displaying incorrect data [...].

Cold Atmospheric Pressure Plasma Comb-A Physical Approach for Pediculosis Treatment.

Pediculosis, that is the infestation of humans with Pediculus humanus capitis (head lice), poses a worldwide problem that is as old as mankind itself. Over the centuries, man has developed a variety of remedies, all of which have ultimately culminated in the use of chemical agents. Some of these remedies are known to produce successful results. A large portion of the effective remedies used to kill lice and their eggs contain insecticides, but there is an increasing number of reports of head lice populations revealing an increased resistance. This study presents an alternative treatment approach, the efficacy of which is based on physical effects. Cold atmospheric pressure plasmas have successfully shown their formidably wide application range within the field of plasma medicine. This study presents a plasma device in its current stage of development that is engineered as a consumer product to enable an alternative physical and insecticide-free option for the treatment of pediculosis. An efficacy study concerning different developmental stages of P. humanus humanus is presented. P. humanus humanus was chosen as a substitute test organism for P. humanus capitis due to possible laboratory rearing and high anatomic similarity. The study shows how a single stroke of the plasma device over a hair strand (approximately 22 cm in length with a weight of 1.5 g) led to mortality rates of 68.3% (50.0; 79.7) (95% CI) in the juvenile test group, a mortality rate of approx. 67.7% (54.9; 78.8) (95% CI) in the female test group, and approx. 46.7% (28.3; 65.7) (95% CI) in the male test group. When single eggs were introduced directly into the plasma for approx. 1 s, younger eggs (0⁻2 d) showed a higher mortality of 66.7% (42.7; 82.7) than the older (4⁻6 d) eggs, with 16.7% (5.6; 34.7) (CI). Furthermore, the results of a risk assessment of the device are described. The article concludes with necessary handling instructions as well as further developmental steps, derived from the results of the efficacy and the risk assessment study.