Search

Terrestrial sedimentary basins are influenced by rapid changes in the clay content, resulting in significant differences in the lateral sealing capacity (shale gouge ratio, SGR) of different parts of the fault. In the present study, we used a method of setting virtual wells and using seismic inversion data to accurately calculate the SGR of faults in strata composed of interbedded sandstone and mudstone calculate the clay content of the strata and to compensate for the low level of the actual well control. Optimal well spacing was determined based on the variable clay content of the formation. The planar variation of the fault throw was characterized via seismic interpretation. We also examined the lateral sealing of faults. The Putaohua oil layer in the S14 area of the Sanzhao Sag in the Songliao Basin was chosen as an case study. Based on the calculation of fault SGR values and the oil distribution, the evaluation criteria for the fault sealing capacity were determined, and the changes in the lateral sealing capacity of the faults were analyzed. This approach accurately estimates fault SGR values and predicts the effective oil-bearing area within the fault zone. It is also suitable for evaluating the lateral sealing of faults in strata composed of interbedded sandstone and mudstone. Our findings provide an in-depth understanding of the lateral sealing of faults and can aid in further research on petroleum distribution patterns.

Biosynthesis of atypical angucyclines involves unique oxidative B-ring cleavage and rearrangement reactions, which are catalyzed by AlpJ-family oxygenases, including AlpJ, JadG, and GilOII. Prior investigations established the essential requirement for FADH2/FMNH2 as cofactors when utilizing the quinone intermediate dehydrorabelomycin as a substrate. In this study, we unveil a previously unrecognized facet of these enzymes as cofactor-independent oxygenases when employing the hydroquinone intermediate CR1 as a substrate. The enzymes autonomously drive oxidative ring cleavage and rearrangement reactions of CR1, yielding products identical to those observed in cofactor-dependent reactions of AlpJ-family oxygenases. Furthermore, the AlpJ- and JadG-catalyzed reactions of CR1 could be quenched by superoxide dismutase, supporting a catalytic mechanism wherein the substrate CR1 reductively activates molecular oxygen, generating a substrate radical and the superoxide anion O2•−. Our findings illuminate a substrate-controlled catalytic mechanism of AlpJ-family oxygenases, expanding the realm of cofactor-independent oxygenases. Notably, AlpJ-family oxygenases stand as a pioneering example of enzymes capable of catalyzing oxidative reactions in either an FADH2/FMNH2-dependent or cofactor-independent manner.

We confirm the previously revised stereochemistry of spiroviolene by X-ray crystallographically characterizing a hydrazone derivative of 9-oxospiroviolane, which is synthesized by hydroboration/oxidation of spiroviolene followed by oxidation of the resultant hydroxy group. An unexpected thermal boron migration occurred during the hydroboration process of spiroviolene that resulted in the production of a mixture of 1α-hydroxyspiroviolane, 9α- and 9β-hydroxyspiroviolane after oxidation. The assertion of the cis-orientation of the 19- and 20-methyl groups provided further support for the revised cyclization mechanism of spiroviolene.

Abstract The biosynthetic investigations of microbial natural products continuously provide powerful biocatalysts for the preparation of valuable chemicals. Practical methods for preparing (S)‐3‐aminopiperidine‐2,6‐dione (2), the pharmacophore of thalidomide (1) and its analog drugs, are highly desired. To develop a biocatalyst for producing (S)‐2, we dissected the domain functions of IdgS, which is responsible for the biosynthesis of indigoidine (3), a microbial blue pigment that consists of two 2‐like moieties. Our data supported that the L‐glutamine tethered to the indigoidine assembly line is first offloaded and cyclized by the thioesterase domain to form (S)‐2, which is then dehydrogenated by the oxidation (Ox) domain and finally dimerized to yield 3. Based on this, we developed an IdgS‐derived enzyme biocatalyst, IdgS‐Ox* R539A, for preparing enantiomerically pure (S)‐2. As a proof of concept, one‐pot chemoenzymatic synthesis of 1 was achieved by combining the biocatalytic and chemical approaches.

Glycerol is a readily available and low-cost simple polyol compound, which can be used as a carbon source for microorganisms to produce various value-added products. Understanding the underlying regulatory mechanism in glycerol metabolism is critical for making better use of glycerol for diverse applications. In a few reported Streptomyces strains, the glycerol utilization gene cluster (glp operon) was shown to be regulated by the IclR family transcriptional regulator GylR. However, the molecular regulatory mechanism mediated by GylR has not been fully elucidated. In this study, we first analyzed the available Actinobacteria genomes in the NCBI Genome database, and found that the glp operon-like gene clusters are conserved in Streptomyces and several other genera of Actinobacteria. By taking Streptomyces clavuligerus NRRL 3585 as a model system, we identified that GylR represses the expressions of glp operon and gylR by directly binding to their promoter regions. Both glycerol-3-phosphate and dihydroxyacetone phosphate can induce the dissociation of GylR from its binding sequences. Furthermore, we identified a minimal essential operator site (a palindromic 18-bp sequence) of GylR-like regulators in Streptomyces. Our study for the first time reported the binding sequences and effector molecules of GylR-like proteins in Streptomyces. The molecular regulatory mechanism mediated by GylR presumably exists widely in Streptomyces. Our findings would facilitate the design of glycerol utilization pathways for producing valuable products. Moreover, our study provided new basic elements for the development of glycerol-inducible regulatory tools for synthetic biology research in the future.

Transcription factor ELONGATED HYPOCOTYL5 (HY5) is the central hub for seedling photomorphogenesis. E3 ubiquitin (Ub) ligase CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) inhibits HY5 protein accumulation through ubiquitination. However, the process of HY5 deubiquitination, which antagonizes E3 ligase-mediated ubiquitination to maintain HY5 homeostasis has never been studied. Here, we identified that Arabidopsis thaliana deubiquitinating enzyme, Ub-SPECIFIC PROTEASE 14 (UBP14) physically interacts with HY5 and enhances its protein stability by deubiquitination. The da3-1 mutant lacking UBP14 function exhibited a long hypocotyl phenotype, and UBP14 deficiency led to the failure of rapid accumulation of HY5 during dark to light. In addition, UBP14 preferred to stabilize nonphosphorylated form of HY5 which is more readily bound to downstream target genes. HY5 promoted the expression and protein accumulation of UBP14 for positive feedback to facilitate photomorphogenesis. Our findings thus established a mechanism by which UBP14 stabilizes HY5 protein by deubiquitination to promote photomorphogenesis in A. thaliana. [ABSTRACT FROM AUTHOR]

Bacterial aromatic polyketides are mainly biosynthesized by type II polyketide synthases (PKSs). The PKSs cannot be functional unless their acyl carrier proteins (ACPs) are phosphopantetheinylated by phosphopantetheinyl transferases (PPTases). Gra-ORF32 was identified as an in-cluster PPTase dedicated for granaticin biosynthesis in Streptomyces vietnamensis and the Arg- and Pro-rich N terminus was found to be crucial for catalytic activity. Overexpression of the encoding genes of the holo-ACP synthases of fatty acid synthases (FAS ACPSs) of both E. coli and S. vietnamensis could efficiently activate the production of granaticins in the Δgra-orf32 mutant, suggesting the ACP of granaticin (graACP) is an efficient substrate for FAS ACPSs. However, Gra-ORF32, the cognate PPTase of the graACP, could not compensate the conditional deficiency of ACPS in E. coli HT253, indicating that it has evolved to be functionally segregated from fatty acid biosynthesis. Nine out of eleven endogenous and all the tested exogenous non-cognate PPTases could activate the production of granaticins to varied extents when overexpressed in the Δgraorf32 mutant, indicating that ACPs of type II PKSs could also be widely recognized as effective substrates by the Sfp-type PPTases. The exogenous PPTases of type II PKSs activated the production of granaticins with much higher efficiency, suggesting that the phylogenetically distant in-cluster PPTases of type II PKSs could share substrate preferences for the ACPs of type II PKSs. A significantly elevated production of granaticins was observed when the mutant Δgra-orf32 was cultivated on ISP2 plates, which was a consequence of crosstalk between the granaticin pathway and a kinamycinlike pathway as revealed by transcriptome analysis and pathway inactivations. Although the host FAS ACPS could efficiently activate the production of granaticins when overexpressed, only Gra-ORF32 activated the efficient production of granaticins under natural physiological conditions, indicating that the activity of the host FAS ACPS was strictly regulated, possibly by binding the FAS holo-ACP product with high affinity. Our findings would contribute to a more comprehensive understanding of how the ACPs of type II PKSs are activated and facilitate the future functional reconstitutions of type II PKSs in E. coli. [ABSTRACT FROM AUTHOR]

Silicone rubber housings crack under the combined internal and external factors and expose the core rod. The exposed core rod deteriorates or breaks under electric fields and electrical current. Based on the main characteristics and mechanism of the core rod corrosion, this paper performs an electrical erosion test in an acid-fog environment and analyzes the core rod's macroscopic results and physicochemical properties. The results indicate the following. First, the discharge on core rod during the electrical erosion process contains four consecutive stages. The electrical current, partial discharge, temperature increase, and macroscopic morphology show regular changes at different stages. The arc usually occurs in the carbonization channel, making the channel have the maximum heat. The area of the core rod heating constantly changes but mainly at the high-voltage side and presents as a regional temperature increase. Second, the fracture of glass fiber and ablation of epoxy resin matrix are related to the continuous action of nitric acid. The generation, expansion, and release of pyrolysis gas form bubbles and pores on the epoxy resin matrix. Erosion and pyrolysis cause the loss of the epoxy resin matrix. Third, the combined effect of electrical current, nitric acid, and heat damage the core rod axially and radially. The study provides support for the subsequent core rod deterioration and fracture mechanism.

Due to excellent anti-pollution performance, composite insulators have been widely used in transmission lines. However, composite insulators present various failures after long-term outdoor operation under environmental, mechanical, and electrical stress. This paper aims to investigate the mechanical deterioration of silicone rubber exposed to electrical stress in an acidic corrosive environment. This study examines the mechanical properties before and after experiments and uses physicochemical properties to analyze aging causes and mechanisms. The results are as follows: a) Subjected to electrical stress in acid mist, alumina tri-hydrate (ATH) decomposition and polydimethylsiloxane (PDMS) pyrolysis occur on the silicone rubber, and oxidative crosslinking reactions take place. The contents of highly oxidized silicon and oxygen increase, and carbon content decreases. The surface discharge aggravates the above progress. b) The electric field has no obvious effect on the penetration of nitric acid into silicone rubber. The surface discharge becomes intense with the increased energized voltage, resulting in decreased hydrophobicity and degraded surface properties. c) Mechanical properties of silicone rubber exposed to acid conditions continuously degrade. Physical crosslinking of the silicone rubber increases along with corrosion time and nitric acid concentration, increasing hardness, decreasing tensile strength, and elongation at break. This paper investigates the relationship between mechanical and physicochemical properties, and the results can provide references for studying the failure mechanism of silicone rubber in an acid corrosion environment.

For the benefit of knowing the method of insulator deicing using hot water, this paper investigated the physical aspects of propagation and the spatial temperature distribution of a hot water jet in the air. The numerical calculation model for the water jet flow field was established and validated, and the temperature spatial variation rule and influencing factors of the water jet were analyzed. The results indicate that the water column breaks as the distance increases. The numerical calculation results and the experimental results of the water jet flow field were approximated. As the distance increases, the mass entrainment rate of the water jet gradually increases, and the normalized axis velocity decreases approximately linearly. The jet temperature rises with the rise in the initial temperature of the hot water, and the rising rate decreases with the increase in distance. The temperature of the water jet falls as the jet distance increases. With the rise in the outlet pressure, the temperature of the water jet drops slightly. The influence of environmental wind speed on the water jet temperature is more significant than ambient temperature—the water jet temperature increases as the nozzle diameter increases. The thorough deicing efficiency is most outstanding when the hot water output temperature is around 86 °C and the jet pressure is ~3.5 MPa.