Hydroxysteroid dehydrogenases (HSDHs) are valuable biocatalysts for the regio‐ and stereoselective modification of steroids, bile acids and other steroid derivatives. In this work, we investigated the substrate promiscuity of this highly selective class of enzymes. In order to reach this goal, a preliminary search of HSDH homologues in in‐house or public available (meta)genomes was carried out. Eight novel NAD(H)‐dependent HSDHs, showing either 7α‐, 7β‐, or 12α‐HSDH activity, and including, for the first time, enzymes from extremophilic microorganisms, were identified, recombinantly produced, and characterized. Among the novel HSDHs, four highly active (up to 92 U mg−1) NAD(H)‐dependent 7β‐HSDHs showing negligible similarity towards previously described 7β‐HSDHs, were discovered. These enzymes, along with previously characterized HSDHs, were tested as biocatalysts for the stereoselective reduction of a panel of substrates including two α‐ketoesters of pharmaceutical interest and selected ketones that partially resemble the structural features of steroids. All the reactions were coupled with a suitable cofactor regeneration system. Regarding the α‐ketoesters, nearly all of the tested HSDHs showed a good activity toward the selected substrates, yielding the reduced α‐hydroxyester with up to 99% conversions and enantiomeric excesses. On the other hand, only the 7β‐HSDHs from Collinsella aerofaciens and Clostridium absonum showed appreciable activity toward more complex ketones, i. e., (±)‐trans‐1‐decalone, but with interesting as well as different selectivity.
The aim of this study was to investigate whether volleyball-specific skills, physical performance, and general cognitive functions differ between players of different competitive levels. Twenty-six female volleyball players competing at 2 different levels (n 5 13, regional; n 5 13, provincial) were tested on volleyball-specific skills (accuracy and technique of setting, passing, spiking, and serving), change of direction speed (COD) by the modified T-test, countermovement jump (CMJ) and general cognitive functions (executive control by Flanker task and perceptual speed by visual search task). Four machine learning models were tested to detect the best one to predict players' level. Regional players presented higher passing, spiking, serving accuracy (p , 0.05) and setting, passing, spiking, and serving technique (p , 0.05) than provincial players. Regional players had also better performance in COD and CMJ than provincial players (p , 0.05). Regional players presented lower response time than provincial players in both congruent and incongruent conditions of the Flanker task, and in both 10 items and 15 items conditions of the visual search task (p , 0.05). Decision tree classifier was the machine learning model with the highest performance to discriminate regional and provincial players (93% precision and 73% recall) by considering passing technique, congruent and incongruent condition of the Flanker task, 15 items and 10 items condition of the visual search task, and spiking technique. These findings demonstrated the importance of assessing volleyball-specific skills and cognitive functions as playing a role to discriminate players of different competitive levels.
The performances of the unspecific peroxigenase from Agrocybe aegerita (AaeUPO) in the asymmetric sulfoxidation of substituted aryl alkyl sulfides were here investigated. A small library of differently substituted aryl alkyl sulfoxides was successfully synthesized from the corresponding sulfides in the presence of AaeUPO and H2O2. All the sulfoxides were obtained as (R)‐enantiomers, regardless the substitution pattern both on the aromatic ring and the alkyl chain, in up to > 99 % conversion and > 99 % ee. An overview about the biocatalytic entries to chiral sulfoxides is also presented here in form of a comparison between the results obtained with AaeUPO and performances of the chloroperoxidase from Caldariomyces fumago, and three different Baeyer–Villiger monooxygenases. To the best our knowledge, this is the first example of a systematic investigation of the AaeUPO synthetic potential in the asymmetric oxidation of hetero atoms, i.e., the pro‐stereogenic sulfur of sulfides.
We report on the covalent immobilization of the (S)‐selective amine transaminase from Vibrio fluvialis (Vf‐ATA) and its use in the synthesis of (S)‐1‐(5‐fluoropyrimidin‐2‐yl)‐ethanamine, a key intermediate of the JAK2 kinase inhibitor AZD1480. Immobilized Vf‐ATA on glyoxyl‐agarose (activity recovery: 30 %) was used in a packed‐bed reactor to set‐up a continuous flow biotransformation coupled with a straightforward in‐line purification to circumvent the 2‐step process described in literature for the batch reaction. The newly developed biotransformation was run in a homogeneous system including dimethyl carbonate as a green co‐solvent. Optically pure (S)‐1‐(5‐fluoropyrimidin‐2‐yl)‐ethanamine (ee >99 %) was isolated in 35 % yield.
Ginger is among the most widespread and widely consumed traditional medicinal plants around the world. Its beneficial effects, which comprise e. g. anticancer and anti‐inflammatory activities as well as gastrointestinal regulatory effects, are generally attributed to a family of non‐volatile compounds characterized by an arylalkyl long‐chained alcohol, diol, or ketone moiety. In this work, ginger active components have been successfully recovered from industrial waste biomass of fermented ginger. Moreover, their recovery has been combined with the first systematic study of the stereoselective reduction of gingerol‐like compounds by isolated alcohol dehydrogenases (ADHs), obtaining the enantioenriched sec‐alcohol derivatives via a sustainable biocatalytic path in up to >99 % conversions and >99 % enantiomeric/diastereomeric excesses.
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