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Endoperoxide-containing natural products are a group of compounds with structurally unique cyclized peroxide moieties. Although numerous endoperoxide-containing compounds have been isolated, the biosynthesis of the endoperoxides remains unclear. NvfI from Aspergillus novofumigatus IBT 16806 is an endoperoxidase that catalyzes the formation of fumigatonoid A in the biosynthesis of novofumigatonin. Here, we describe our structural and functional analyses of NvfI. The structural elucidation and mutagenesis studies indicate that NvfI does not utilize a tyrosyl radical in the reaction, in contrast to other characterized endoperoxidases. Further, the crystallographic analysis reveals significant conformational changes of two loops upon substrate binding, which suggests a dynamic movement of active site during the catalytic cycle. As a result, NvfI installs three oxygen atoms onto a substrate in a single enzyme turnover. Based on these results, we propose a mechanism for the NvfI-catalyzed, unique endoperoxide formation reaction to produce fumigatonoid A. Many endoperoxide-containing natural products have been isolated, but the biosynthesis of the endoperoxides remains unclear. Here, the authors report the structural and functional analysis of the NvfI endoperoxidase that catalyzes the formation of fumigatonoid A in the biosynthesis of novofumigatonin, and show that it does not employ tyrosyl radical in the reaction.

This study alleviates the low operating temperature constraint of Si qubits. A qubit is a key element for quantum sensors, memories, and computers. Electron spin in Si is a promising qubit, as it allows both long coherence times and potential compatibility with current silicon technology. Si qubits have been implemented using gate-defined quantum dots or shallow impurities. However, operation of Si qubits has been restricted to milli-Kelvin temperatures, thus limiting the application of the quantum technology. In this study, we addressed a single deep impurity, having strong electron confinement of up to 0.3 eV, using single-electron tunnelling transport. We also achieved qubit operation at 5–10 K through a spin-blockade effect based on the tunnelling transport via two impurities. The deep impurity was implemented by tunnel field-effect transistors (TFETs) instead of conventional FETs. With further improvement in fabrication and controllability, this work presents the possibility of operating silicon spin qubits at elevated temperatures.

New enzyme catalysts are usually engineered by repurposing the active sites of natural proteins. Here we show that design and directed evolution can be used to transform a non-natural, functionally naive zinc-binding protein into a highly active catalyst for an abiological hetero-Diels–Alder reaction. The artificial metalloenzyme achieves >104 turnovers per active site, exerts absolute control over reaction pathway and product stereochemistry, and displays a catalytic proficiency (1/KTS = 2.9 × 1010 M−1) that exceeds all previously characterized Diels–Alderases. These properties capitalize on effective Lewis acid catalysis, a chemical strategy for accelerating Diels–Alder reactions common in the laboratory but so far unknown in nature. Extension of this approach to other metal ions and other de novo scaffolds may propel the design field in exciting new directions.A de novo designed zinc-binding protein has been converted into a highly active, stereoselective catalyst for a hetero-Diels–Alder reaction. Design and directed evolution were used to effectively harness Lewis acid catalysis and create an enzyme more proficient than other reported Diels–Alderases.

Non-heme iron and α-ketoglutarate-dependent (Fe/αKG) oxygenases catalyze various oxidative biotransformations. Due to their catalytic flexibility and high efficiency, Fe/αKG oxygenases have attracted keen attention for their application as biocatalysts. Here, we report the biochemical and structural characterizations of the unusually promiscuous and catalytically versatile Fe/αKG oxygenase SptF, involved in the biosynthesis of fungal meroterpenoid emervaridones. The in vitro analysis revealed that SptF catalyzes several continuous oxidation reactions, including hydroxylation, desaturation, epoxidation, and skeletal rearrangement. SptF exhibits extremely broad substrate specificity toward various meroterpenoids, and efficiently produced unique cyclopropane-ring-fused 5/3/5/5/6/6 and 5/3/6/6/6 scaffolds from terretonins. Moreover, SptF also hydroxylates steroids, including androsterone, testosterone, and progesterone, with different regiospecificities. Crystallographic and structure-based mutagenesis studies of SptF revealed the molecular basis of the enzyme reactions, and suggested that the malleability of the loop region contributes to the remarkable substrate promiscuity. SptF exhibits great potential as a promising biocatalyst for oxidation reactions. Non-heme iron and α-ketoglutarate-dependent (Fe/αKG) oxygenases have attracted attention for their application as biocatalysts due to their flexibility and high efficiency. Here, the authors show the biochemical and structural characterizations of the versatile Fe/αKG oxygenase SptF, involved in the biosynthesis of fungal meroterpenoid emervaridones.

A coronary bifurcation stenting is still a challenging issue due to frequent restenosis and stent thrombosis even with drug-eluting stents. The bifurcation angle (BA) between a main vessel and a side branch is one of the crucial determinants of coronary flow and shear stress that affect the plaque distribution. Previous bench and clinical studies have evaluated the impact of the BA between the proximal main vessel and the side branch (Angle A) and the BA between the distal main vessel and the side branch (Angle B) on the clinical outcomes of bifurcation stenting. However, the impact has not yet been fully elucidated due to a lack of statistical power or different manner of the assessment of BA. To analyze the published studies on coronary artery BA, the modalities used for assessment, and the impact of BA on interventions and attempt to define the pre-procedural protocols. A scoping review was performed using the Joanna Briggs Institute Methodology. A total of 52 relevant references were selected from PubMed, Cochrane Library, and CINAHL databases and categorized into three topic areas. A wider Angle A is associated with the increased likelihood of carina shift and a wider Angle B, with that of side branch occlusion. A wider Angle B promotes stent malapposition and deformation in the side branch ostium and has been reported as an independent predictor of major adverse cardiac events after bifurcation stenting; however, improvement of the drug-eluting stent, refinement of the stenting technique, and accurate 3-dimensional assessment may attenuate the adverse clinical impact of a wider BA. Assessment of the BA is necessary to predict the effect of bifurcation intervention procedure on the stent configuration and coronary flow at the bifurcated vessels. This will help to optimize stent selection and the stenting technique.

C -Glycosides, in which a sugar moiety is linked via a carbon-carbon (C-C) bond to a non-sugar moiety (aglycone), are found in our food and medicine. The C-C bond is cleaved by intestinal microbes and the resulting aglycones exert various bioactivities. Although the enzymes responsible for the reactions have been identified, their catalytic mechanisms and the generality of the reactions in nature remain to be explored. Here, we present the identification and structural basis for the activation of xenobiotic C -glycosides by heterocomplex C -deglycosylation enzymes from intestinal and soil bacteria. They are found to be metal-dependent enzymes exhibiting broad substrate specificity toward C -glycosides. X-ray crystallographic and cryo-electron microscopic analyses, as well as structure-based mutagenesis, reveal the structural details of these enzymes and the detailed catalytic mechanisms of their remarkable C-C bond cleavage reactions. Furthermore, bioinformatic and biochemical analyses suggest that the C -deglycosylation enzymes are widely distributed in the gut, soil, and marine bacteria. In C-glycosides the sugar moiety is linked through a carbon-carbon bond to the non-sugar moiety, which can be cleaved by intestinal microbes. Here, the authors use bioinformatics analysis to identify C-glycoside deglycosidase enzymes in intestinal and soil bacteria, biochemically characterise them and determine their structures and probe catalytic important residues in mutagenesis experiments.

2-(2-Phenylethyl)chromones (PECs) are the principal constituents contributing to the distinctive fragrance of agarwood. How PECs are biosynthesized is currently unknown. In this work, we describe a diarylpentanoid-producing polyketide synthase (PECPS) identified from Aquilaria sinensis . Through biotransformation experiments using fluorine-labeled substrate, transient expression of PECPS in Nicotiana benthamiana , and knockdown of PECPS expression in A. sinensis calli, we demonstrate that the C 6 –C 5 –C 6 scaffold of diarylpentanoid is the common precursor of PECs, and PECPS plays a crucial role in PECs biosynthesis. Crystal structure (1.98 Å) analyses and site-directed mutagenesis reveal that, due to its small active site cavity (247 Å 3 ), PECPS employs a one-pot formation mechanism including a “diketide-CoA intermediate-released” step for the formation of the C 6 –C 5 –C 6 scaffold. The identification of PECPS, the pivotal enzyme of PECs biosynthesis, provides insight into not only the feasibility of overproduction of pharmaceutically important PECs using metabolic engineering approaches, but also further exploration of how agarwood is formed. 2-(2-Phenylethyl)chromones (PECs) contribute to the distinctive fragrance of agarwood. Here the authors identify a diarylpentanoid-producing polyketide synthase from Aquilaria sinensis and show how it catalyzes PEC formation.

Endoperoxide natural products are widely distributed in nature and exhibit various biological activities. Due to their chemical features, endoperoxide and endoperoxide-derived secondary metabolites have attracted keen attention in the field of natural products and organic synthesis. In this review, we summarize the structural analyses, mechanistic investigations, and proposed reaction mechanisms of endoperoxide-forming oxygenases, including cyclooxygenase, fumitremorgin B endoperoxidase (FtmOx1), and the asnovolin A endoperoxygenase NvfI.

All known triterpenes are generated by triterpene synthases (TrTSs) from squalene or oxidosqualene 1 . This approach is fundamentally different from the biosynthesis of short-chain (C 10 –C 25 ) terpenes that are formed from polyisoprenyl diphosphates 2 – 4 . In this study, two fungal chimeric class I TrTSs, Talaromyces verruculosus talaropentaene synthase (TvTS) and Macrophomina phaseolina macrophomene synthase (MpMS), were characterized. Both enzymes use dimethylallyl diphosphate and isopentenyl diphosphate or hexaprenyl diphosphate as substrates, representing the first examples, to our knowledge, of non-squalene-dependent triterpene biosynthesis. The cyclization mechanisms of TvTS and MpMS and the absolute configurations of their products were investigated in isotopic labelling experiments. Structural analyses of the terpene cyclase domain of TvTS and full-length MpMS provide detailed insights into their catalytic mechanisms. An AlphaFold2-based screening platform was developed to mine a third TrTS, Colletotrichum gloeosporioides colleterpenol synthase (CgCS). Our findings identify a new enzymatic mechanism for the biosynthesis of triterpenes and enhance understanding of terpene biosynthesis in nature. Chimeric triterpene synthases are identified that catalyse non-squalene-dependent triterpene biosynthesis.

Background: This study aimed to determine whether there are disparities in healthcare services utilization according to household income among people aged 75 years or older in Japan. Methods: We used data on medical and long-term care (LTC) insurance claims and on LTC insurance premiums and needs levels for people aged 75 years or older in a suburban city. Data on people receiving public welfare were not available. Participants were categorized according to household income level using LTC insurance premiums data. The associations of low income with physician visit frequency, length of hospital stay (LOS), and medical and LTC expenditures were evaluated and adjusted for 5-year age groups and LTC needs level. Results: The study analyzed 12,852 men and 18,020 women, among which 13.3% and 41.5%, respectively, were categorized as low income. Participants with low income for both genders were more likely to be functionally dependent. In the adjusted analyses, lower income was associated with fewer physician visits (incidence rate ratio [IRR] 0.90; 95% confidence interval [CI], 0.87–0.92 for men and IRR 0.97; 95% CI, 0.95–0.99 for women), longer LOS (IRR 1.98; 95% CI, 1.54–2.56 and IRR 1.42; 95% CI, 1.20–1.67, respectively), and higher total expenditures (exp(β) 1.09; 95% CI, 1.01–1.18 and exp(β) 1.09; 95% CI, 1.05–1.14, respectively). Conclusions: This study suggests that older people with lower income had fewer consultations with physicians but an increased use of inpatient services. The income categorization used in this study may be an appropriate proxy of socioeconomic status.