Enzyme In Biosynthesis Research Articles

The UV-A Receptor CRY-DASH1 Up- and Downregulates Proteins Involved in Different Plastidial Pathways

Algae encode up to five different types of cryptochrome photoreceptors. So far, relatively little is known about the biological functions of the DASH (Drosophila, Arabidopsis, Synechocystis and Homo)-type cryptochromes. The green alga Chlamydomonas reinhardtii encodes two of them. CRY-DASH1 also called DCRY1 has its maximal absorption peak in the UV-A range. It is localized in the chloroplast and plays an important role in balancing the photosynthetic machinery. Here, we performed a comparative analysis of chloroplast proteins from wild type and a knockout mutant of CRY-DASH1 named cry-dash1mut, using label-free quantitative proteomics as well as immunoblotting. Our results show upregulation of enzymes involved in specific pathways in the mutant including key enzymes of chlorophyll and carotenoid biosynthesis consistent with increased levels of photosynthetic pigments in cry-dash1mut. There is also an increase in certain redox as well as photosystem I and II proteins, including D1. Strikingly, CRY-DASH1 is coregulated in a D1 deletion mutant, where its amount is increased. In contrast, key proteins of the central carbon metabolism, including glycolysis/gluconeogenesis, dark fermentation and the oxidative pentose phosphate pathway are downregulated in cry-dash1mut. Similarly, enzymes of histidine biosynthesis are downregulated in cry-dash1mut leading to a reduction in the amount of free histidine. Yet, transcripts encoding for several of these proteins are at a similar level in the wild type and cry-dash1mut or even opposite. We show that CRY-DASH1 can bind to RNA, taking the psbA RNA encoding D1 as target. These data suggest that CRY-DASH1 regulates plastidial metabolic pathways at the posttranscriptional level.

EXPRESSION OF DE NOVO CERAMIDE SYNTHESIS IN LOCAL HEART FAT DEPO IN PATIENTS WITH CARDIOVASCULAR DISEASES

Introduction. Ceramides are bioactive lipids that are not only structural components of cell membranes, but also second messengers in the transmission of cell signals. Ceramides are associated with risk factors for cardiovascular disease, may be independent predictors of cardiovascular events and proatherogenic mediators. However, due attention has not been paid to the problem of ceramide accumulation in the adipose tissue (AT) of the heart, while the accumulation of ceramides in adipocytes of epicardial and perivascular AT in coronary heart disease (CHD) is of particular interest, since they are localized in close proximity to the coronary vessels and the focus of the lesion. Objective: to evaluate the expression of de novo ceramide biosynthesis enzymes in the adipose tissue of the heart of various locations in patients with coronary heart disease and acquired heart defects. Materials and methods. The study included 30 patients with CAD, the comparison group consisted of 30 patients with acquired valvular disease (aortic stenosis/insufficiency) without CAD. During the operation, biopsies of subcutaneous, epicardial, perivascular adipose tissue (VT) (SAT, EAT, PVAT, respectively) were obtained. Expression of genes encoding de novo ceramide synthesis enzymes (subunits of C1 and C2 serine palmitoyltransferases: SPTLC1, SPTLC2; ceramide synthase 1-6: CERS1-6; dihydroceramide desaturase: DEGS1) was assessed by real-time quantitative polymerase chain reaction (qPCR) with primers synthesized by Evrogen (Moscow, Russia) on the ViiA 7 Real-Time PCR System (Applied Biosystems, Foster City, California, USA). Statistical analysis of the results was performed using GraphPad Prism 8 (GraphPad Software). Results. Patients with CAD were characterized by higher levels of SPTLC1 mRNA in subcutaneous and epicardial VT and SPTLC2 mRNA in EAT. DEGS1 expression was maximal in SVT and EAT. The expression of ceramide synthase genes in VT of patients with coronary artery disease had tissue-specific features. Expression of the CERS1 gene, encoding the enzyme ceramide synthase-1, which produces ceramides with a fatty acid residue in its composition of 18 carbon atoms (C18), expression of CERS5 and CERS6, encoding ceramide synthase-5 and -6, synthesizing ceramides C14-C16 was maximum in epicardial adipocytes. At the same time, CERS5 expression was higher than CERS6 expression. Gene expression of CERS2, encoding the enzyme ceramide synthase-2, which produces long-chain ceramides and attaches C20–C24 fatty acyl-CoA to the sphingoid base in ceramide, was maximal in subcutaneous adipocytes. While perivascular VT was characterized by a higher expression of the CERS4 and CERS3 genes, which synthesize very long chain ceramides C26 and higher. In the group of patients with valvular disease, VT samples did not differ in terms of SPTLC1, SPTLC2, CERS1, CERS2, CERS5, and CERS6 mRNA levels. However, high expression of CERS4 and DEGS1 was observed in EAT, while CERS3 and CERS4 were expressed in PVAT. When comparing the level of expression of genes for ceramide synthesis enzymes between the two studied groups, it was found that in patients with IHD, the mRNA level of SPTLC1 in SAT and EAT, SPTLC2 in EAT, CERS2 in all types of AT, CERS4 and CERS5 in EAT, DEGS1 in SAT and EAT was higher in compared with patients with valvular heart disease. Conclusion. Regional fat depots of the heart differed in the level of expression of de novo ceramide biosynthesis enzymes. The results obtained indicate the activation of ceramide synthesis along this pathway in adipocytes of predominantly epicardial localization in coronarogenic pathology, which may contribute to the accumulation of long-chain ceramides in the VT of this localization.

Serological and molecular typing of Tenacibaculum maritimum from New Zealand farmed salmon, Oncorhynchus tshawytscha

Tenacibaculum maritimum is a cosmopolitan bacterial pathogen with the potential to cause significant losses in a broad range of farmed and wild marine fish species. This study investigated the antigenic diversity of T. maritimum isolated in culture from farmed Chinook (king) salmon (Oncorhynchus tshawytscha) from the Marlborough Sounds in New Zealand. A total of 36 isolates were examined using antibody serotyping and rapid molecular serotyping via multiplex PCR (mPCR) targeting genes encoding O-antigen biosynthesis enzymes. Serological analysis using three different polyclonal antisera developed against T. maritimum isolated from farmed Atlantic salmon (Salmo salar) in Tasmania, Australia revealed that there are three putative serotypes of T. maritimum that occur in New Zealand. The predominant serotype was defined by a positive reaction to all three Tasmanian antisera (antisera A, B and C), designated as serotype ABC. This serotype was found at all nine farm locations tested and represented 81% of all isolates examined. The same library of isolates was evaluated by mPCR serotyping and found three O-AGC types among tested isolates. O-AGC Type 3 was not only the predominant type (72%) present, but it also had a wide distribution, having been isolated at eight of the nine farms. Two other O-AGC types (O-AGC Type 2–1 and O-AGC Type 3–2) were identified, providing evidence of genetic variation. However, there was only partial concordance between the two serotyping techniques, which is likely linked to differences in the way serotypes are defined in the two approaches that were used. Nevertheless, in broad terms there is good evidence of intraspecific antigenic variation within our library of isolates, and collectively these data will be of crucial importance for assessing the pathogenicity of the isolates and the subsequent development of a vaccine for this emerging disease in New Zealand marine salmon farms.

A 3-Ketoacyl-CoA Synthase 10 (KCS10) Homologue from Alfalfa Enhances Drought Tolerance by Regulating Cuticular Wax Biosynthesis.

Cuticular wax, forming the first line of defense against adverse environmental stresses, comprises very long-chain fatty acids (VLCFAs) and their derivatives. 3-Ketoacyl-CoA synthase (KCS) is a rate-limiting enzyme for VLCFA biosynthesis. In this study, we isolated KCS10, a KCS gene from alfalfa, and analyzed the effect of gene expression on wax production and drought stress in transgenic plants. MsKCS10 overexpression increased compact platelet-like crystal deposition and promoted primary alcohol biosynthesis through acyl reduction pathways in alfalfa leaves. Overexpression of MsKCS10 induced the formation of coiled-rodlet-like crystals and increased n-alkane content through decarbonylation pathways in tobacco and tomato fruits. Overexpression of MsKCS10 enhanced drought tolerance by limiting nonstomatal water loss, improving photosynthesis, and maintaining osmotic potential under drought stress in transgenic tobacco. In summary, MsKCS10 plays an important role in wax biosynthesis, wax crystal morphology, and drought tolerance, although the mechanisms are different among the plant species. MsKCS10 can be targeted in future breeding programs to improve drought tolerance in plants.

Subsets of preoperative sex hormones in testicular germ cell cancer: a retrospective multicenter study

Preoperative homeostasis of sex hormones in testicular germ cell tumor (TGCT) patients is scarcely characterized. We aimed to explore regulation of sex hormones and their implications for histopathological parameters and prognosis in TGCT using a data-driven explorative approach. Pre-surgery serum concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), estradiol (E2) and prolactin were measured in a retrospective multicenter TGCT cohort (n = 518). Clusters of patients were defined by latent class analysis. Clinical, pathologic and survival parameters were compared between the clusters by statistical hypothesis testing, Random Forest modeling and Peto-Peto test. Cancer tissue expression of sex hormone-related genes was explored in the publicly available TCGA cohort (n = 149). We included 354 patients with pure seminoma and 164 patients with non-seminomatous germ cell tumors (NSGCT), with a median age of 36 years. Three hormonal clusters were defined: ‘neutral’ (n = 228) with normal sex hormone homeostasis, ‘testicle’ (n = 91) with elevated T and E2, low pituitary hormones, and finally ‘pituitary’ subset (n = 103) with increased FSH and LH paralleled by low-to-normal levels of the gonadal hormones. Relapse-free survival in the hormonal subsets was comparable (p = 0.64). Cancer tissue expression of luteinizing hormone- and follicle-stimulating hormone-coding genes was significantly higher in seminomas, while genes of T and E2 biosynthesis enzymes were strongly upregulated in NSGCT. Substantial percentages of TGCT patients are at increased risk of sex hormone dysfunction at primary diagnosis before orchiectomy. TGCT may directly influence systemic hormonal homeostasis by in-situ synthesis of sex hormones.

Analysis of CYP17 gene polymorphism on increasing androstenedione levels in polycystic ovary syndrome women with obesity in Surabaya

Introduction: CYP17 gene polymorphism causes susceptibility, can interfere with the enzyme biosynthesis mechanism in steroidogenesis, and becomes a possible genetic factor for hyper-androgen in PCOS. The study aimed to analyze the relationship between CYP17 gene polymorphism and increased androstenedione levels in PCOS women with obesity. Methods: This study is an observational case-control analytic. The sample is 23 obese PCOS women and 23 obese non-PCOS women, using a purposive sampling technique. CYP17 gene polymorphism examination with PCR and RFLP. Examination of androstenedione hormone by ELISA. The examination results were tested using a logistic regression statistical test with α = 0.05, the hypothesis being accepted if p-value <0.005. Results: The study subjects were 20-30 years old, 50-60 kg body weight, with BMI more than 25%. The distribution of CYP17 gene genotypes in the case group was TC, 47.8%, while in the control group, it was TT, 100%, Chi-Square test on CYP17 gene polymorphism in PCOS and control groups showed significant differences (P 0.000). Wilcoxon-Mann Whitney test of androstenedione levels in cases compared to controls P = 0.000, eta CYP17 correlation test with androstenedione levels P = 0.061. Conclusion: There is a difference in CYP 17 gene polymorphism in obese PCOS women compared to obese non-PCOS women. Androstenedione levels are higher in PCOS women than in obese non-PCOS women and there is no association of CYP17 with Androstenedione levels in PCOS women with obesity.

Wheat dihydroflavonol-4-reductase like gene confers disease resistance by modulating NADPH pool and lignin biosynthesis

Hm1 is the first cloned disease resistance gene (R) from maize which encodes an enzyme that detoxifies Helminthosporium carbonum (HC) toxin from the fungal pathogen Cochliobolus carbonum. The homologous genes for Hm1 have been mined in various crop plants and their sequences are showed similar to dihydroflavonol-4-reductase (DFR), a key enzyme in flavonoid biosynthesis, therefore they are collectively named dihydroflavonol-4-reductase like (DFRL). The biological functions of DFRL are largely elusive. In the present study, DFRL gene was cloned from wheat, namely TaDFRL. TaDFRL was barely expressed in leaf, stem and root tissues; however, its expression level was rapidly increased following rust infection. Biochemical analysis showed that TaDFRL had the broad spectrum of substrate preference, including dihydroflavonol, flavonol and flavone, and could use both NAD and NADP as co-enzyme, which was quite distinct from DFR. Overexpressing TaDFRL in tobacco altered NAD(H) and NADP(H) pools towards to high NADPH levels. Subsequently, the gene expression of cinnamyl alcohol dehydrogenase (CAD) was up-regulated and lignin accumulation was increased. These brought about to enhance resistance to wildfire disease in tobacco plants. This research provides novel insights into DFRL mechanism, which boost host defense responses by elevating NADPH level and lignin biosynthesis.

The apocarotenoid production in microbial biofactories: An overview.

Carotenoids are a vast group of natural pigments that come in a variety of colors ranging from red to orange. Apocarotenoids are derived from these carotenoids, which are hormones, pigments, retinoids, and volatiles employed in the textiles, cosmetics, pharmaceutical, and food industries. Due to the high commercial value and poor natural host abundance, they are significantly undersupplied. Microbes like Saccharomyces cerevisiae and Escherichia coli act as heterologous hosts for apocarotenoid production. This article briefly reviews categories of apocarotenoids, their biosynthetic pathway commencing from the MVA and MEP, its significance, the tool enzymes for apocarotenoid biosynthesis like CCDs, their biotechnological production in microbial factories, and future perspectives.

Synthesis and Evaluation of Novel Triaryl Derivatives with Readthrough-Inducing Activity.

The readthrough mechanism, which skips the premature termination codon and restores the biosynthesis of the defective enzyme, is an emerging therapeutic tactic for nonsense mutation-related diseases, such as Hurler syndrome, a type of mucopolysaccharidosis. In the present study, novel triaryl derivatives were synthesized and their readthrough-inducing activities were evaluated by a luciferase reporter assay with a partial α-L-iduronidase (IDUA) DNA sequence containing the Q70X nonsense mutation found in Hurler syndrome and by measuring the enzyme activity of IDUA knockout cells transfected with the mutant IDUA gene. KY-516, a representative compound in which the meta position carboxyl group of the left ring of the clinically used ataluren was converted to the para position sulfamoylamino group, the central ring to triazole, and the right ring to cyanobenzene, exhibited the most potent readthrough-inducing activity in the Q70X/luciferase reporter assay. In Q70X mutant IDUA transgenic cells, KY-516 significantly increased enzyme activity at 0.1 µM. After the oral administration of KY-516 (10 mg/kg), the highest plasma concentration of KY-516 was above 5 µM in rats. These results indicate that KY-516, a novel triaryl derivative, exhibits potent readthrough-inducing activity and has potential as a therapeutic agent for Hurler syndrome.

FACTORS ON THE BIOSYNTHESIS OF AMYLOLITIC ENZYMES OF STREPTOMYCETE ORIGIN

Modern enzyme biotechnology is a promising and rapidly developing field that requires the latest research on the conditions of enzyme biosynthesis. Optimizing the composition of the nutrient medium depending on the needs of microorganisms and physicochemical factors directly affect the increase in the efficiency of the biosynthesis of amylolytic enzymes, namely the biosynthetic capacity of the strain Streptomyces recifensis var. lyticus 2P-15. Modulation of the biosynthetic activity of strains producing amylolytic enzymes will allow to significantly increase their economic yield. Aim. The purpose of this work is to optimize the biosynthetic capacity of the strain Streptomyces recifensis var. lyticus 2P-15 in terms of the synthesis of amylolytic enzymes and the study of the dynamics of the influence of physical and chemical factors on optimization. Methods. The object of the study is the strain Streptomyces recifensis var. lyticus 2P-15, obtained by three-stage selection of the producer. The simplex method of selecting the composition of the environment was used for the research. The ratio of amylolytic activity to the level of biomass accumulation was taken as the biosynthetic capacity of the strain. A photocolometric method was used to determine amylolytic activity. The level of biomass accumulation was determined by the weight method. Results. It was established that as a result of optimizing the composition of the simplex nutrient medium by the method of mathematical modeling, the biosynthetic capacity increased by 3.63 compared to the control variant. It was also investigated that the optimal concentration of such a component of the nutrient medium as monosodium glutamate С5Н8NO4Na・H2O is 1.5%, which increases the amylolytic activity by 2.63 and increases the accumulation of biomass. Separately, it should be noted the obtained results of the study of the optimal concentrations of heavy metal ions added to the optimized version of the nutrient medium, which allow further research in this aspect to be continued and the use of Co, Mo, Cd ions in the composition of the nutrient medium. With the obtained results, there is an increase in amylolytic activity in the best response by 3.54. The obtained results have theoretical and practical significance for further research in the biotechnology of enzymes. Conclusions. The prospect of further research into the optimization of the biosynthesis of actinomycetes by the simplex method of other aspects of its regulation will be to increase the biosynthetic capacity of the studied strain, which will have a positive effect on the economic output of the production of amylolytic enzyme preparations by obtaining microbial synthesis.

Asymmetric Distribution of Plasmalogens and Their Roles-A Mini Review.

Plasmalogens are a unique family of cellular glycerophospholipids that contain a vinyl-ether bond. The synthesis of plasmalogens is initiated in peroxisomes and completed in the endoplasmic reticulum. Plasmalogens are transported to the post-Golgi compartment, including endosomes and plasma membranes, in a manner dependent on ATP, but not vesicular transport. Plasmalogens are preferentially localized in the inner leaflet of the plasma membrane in a manner dependent on P4-type ATPase ATP8B2, that associates with the CDC50 subunit. Plasmalogen biosynthesis is spatiotemporally regulated by a feedback mechanism that senses the amount of plasmalogens in the inner leaflet of the plasma membrane and controls the stability of fatty acyl-CoA reductase 1 (FAR1), the rate-limiting enzyme for plasmalogen biosynthesis. The physiological consequences of such asymmetric localization and homeostasis of plasmalogens are discussed in this review.

Integrated analyses of metabolomics and transcriptomics reveal the potential regulatory roles of long non-coding RNAs in gingerol biosynthesis

BackgroundAs the characteristic functional component in ginger, gingerols possess several health-promoting properties. Long non-coding RNAs (lncRNAs) act as crucial regulators of diverse biological processes. However, lncRNAs in ginger are not yet identified so far, and their potential roles in gingerol biosynthesis are still unknown. In this study, metabolomic and transcriptomic analyses were performed in three main ginger cultivars (leshanhuangjiang, tonglingbaijiang, and yujiang 1 hao) in China to understand the potential roles of the specific lncRNAs in gingerol accumulation.ResultsA total of 744 metabolites were monitored by metabolomics analysis, which were divided into eleven categories. Among them, the largest group phenolic acid category contained 143 metabolites, including 21 gingerol derivatives. Of which, three gingerol analogs, [8]-shogaol, [10]-gingerol, and [12]-shogaol, accumulated significantly. Moreover, 16,346 lncRNAs, including 2,513, 1,225, and 2,884 differentially expressed (DE) lncRNA genes (DELs), were identified in all three comparisons by transcriptomic analysis. Gene ontology enrichment (GO) analysis showed that the DELs mainly enriched in the secondary metabolite biosynthetic process, response to plant hormones, and phenol-containing compound metabolic process. Correlation analysis revealed that the expression levels of 11 DE gingerol biosynthesis enzyme genes (GBEGs) and 190 transcription factor genes (TF genes), such as MYB1, ERF100, WRKY40, etc. were strongly correlation coefficient with the contents of the three gingerol analogs. Furthermore, 7 and 111 upstream cis-acting lncRNAs, 1,200 and 2,225 upstream trans-acting lncRNAs corresponding to the GBEGs and TF genes were identified, respectively. Interestingly, 1,184 DELs might function as common upstream regulators to these GBEGs and TFs genes, such as LNC_008452, LNC_006109, LNC_004340, etc. Furthermore, protein–protein interaction networks (PPI) analysis indicated that three TF proteins, MYB4, MYB43, and WRKY70 might interact with four GBEG proteins (PAL1, PAL2, PAL3, and 4CL-4).ConclusionBased on these findings, we for the first time worldwide proposed a putative regulatory cascade of lncRNAs, TFs genes, and GBEGs involved in controlling of gingerol biosynthesis. These results not only provide novel insights into the lncRNAs involved in gingerol metabolism, but also lay a foundation for future in-depth studies of the related molecular mechanism.

AtMYBS1 negatively regulates heat tolerance by directly repressing the expression of MAX1 required for strigolactone biosynthesis in Arabidopsis

Heat stress caused by global warming requires the development of thermotolerant crops to sustain yield. It is necessary to understand the molecular mechanisms that underlie heat tolerance in plants. Strigolactones (SLs) are a class of carotenoid-derived phytohormones that regulate plant development and responses to abiotic or biotic stresses. Although SL biosynthesis and signaling processes are well established, genes that directly regulate SL biosynthesis have rarely been reported. Here, we report that the MYB-like transcription factor AtMYBS1/AtMYBL, whose gene expression is repressed by heat stress, functions as a negative regulator of heat tolerance by directly inhibiting SL biosynthesis in Arabidopsis. Overexpression of AtMYBS1 led to heat hypersensitivity, whereas atmybs1 mutants displayed increased heat tolerance. Expression of MAX1, a critical enzyme in SL biosynthesis, was induced by heat stress and downregulated in AtMYBS1-overexpression (OE) plants but upregulated in atmybs1 mutants. Overexpression of MAX1 in the AtMYBS1-OE background reversed the heat hypersensitivity of AtMYBS1-OE plants. Loss of MAX1 function in the atmyb1 background reversed the heat-tolerant phenotypes of atmyb1 mutants. Yeast one-hybrid assays, chromatin immunoprecipitation‒qPCR, and transgenic analyses demonstrated that AtMYBS1 directly represses MAX1 expression through the MYB binding site in the MAX1 promoter in vivo. The atmybs1d14 double mutant, like d14 mutants, exhibited hypersensitivity to heat stress, indicating the necessary role of SL signaling in AtMYBS1-regulated heat tolerance. Our findings provide new insights into the regulatory network of SL biosynthesis, facilitating the breeding of heat-tolerant crops to improve crop production in a warming world.

3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase (HMGCR) protects hair cells from cisplatin-induced ototoxicity in vitro: possible relation to the activities of p38 MAPK signaling pathway.

The 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase (HMGCR) gene encodes rate-limiting enzyme in cholesterol biosynthesis, which is related to cell proliferation and mitochondrial function. The present study was designed to explore the expression of HMGCR in murine cochlear hair cells and HEI-OC1 cells and the possible mechanisms underpinning the actions of HMGCR in cisplatin-induced ototoxicity, with special attention given to p38 mitogen-activated protein kinase (MAPK) activities in vitro. The expressions of HMGCR, p-p38, cleaved caspase-3 and LC3B was measured by immunofluorescence and western blot. JC-1 staining and MitoSOX Red were used to detect mitochondria membrane potential (MMP) and reactive oxygen species (ROS) levels respectively. The apoptosis of auditory cells was assessed by TUNEL staining and flow cytometry. Protein levels of bcl2/bax and beclin1 were examined by western blot. We found that HMGCR was widely expressed in the auditory cells, of both neonatal mice and 2-month-old mice, in cytoplasm, nucleus and stereocilia. Moreover, 30μM cisplatin elicited the formation of ROS, which, in turn, led to HMGCR reduction, activating p38 kinase-related apoptosis and autophagy in auditory cells. Meanwhile, co-treatment with ROS scavenger at a concentration of 2mM, N-acetyl-L-cysteine (NAC), could alleviate the aforementioned changes. In addition, HMGCR silencing resulted in higher p38 MAPK-mediated apoptosis and autophagy under cisplatin injury. Taken together, we demonstrate that, for the first time, that HMGCR is expressed in the cochlear. Furthermore, HMGCR exerts protective benefit on auditory cells against cisplatin-mediated injury stimulated by ROS, culminating in regulation of p38 MAPK-dependent apoptosis and autophagy.

Altered bile acid metabolism in skin tissues in response to ionizing radiation: deoxycholic acid (DCA) as a novel treatment for radiogenic skin injury

Objective Radiogenic skin injury (RSI) is a common complication during cancer radiotherapy or accidental exposure to radiation. The aim of this study is to investigate the metabolism of bile acids (BAs) and their derivatives during RSI. Methods Rat skin tissues were irradiated by an X-ray linear accelerator. The quantification of BAs and their derivatives were performed by liquid chromatography–mass spectrometry (LC–MS)-based quantitative analysis. Key enzymes in BA biosynthesis were analyzed from single-cell RNA sequencing (scRNA-Seq) data of RSI in the human patient and animal models. The in vivo radioprotective effect of deoxycholic acid (DCA) was detected in irradiated SD rats. Results Twelve BA metabolites showed significant differences during the progression of RSI. Among them, the levels of cholic acid (CA), DCA, muricholic acid (MCA), chenodeoxycholic acid (CDCA), glycocholic acid (GCA), glycohyodeoxycholic acid (GHCA), 12-ketolithocholic acid (12-ketoLCA) and ursodeoxycholic acid (UDCA) were significantly elevated in irradiated skin, whereas lithocholic acid (LCA), tauro-β-muricholic acid (Tβ-MCA) and taurocholic acid (TCA) were significantly decreased. Additionally, the results of scRNA-Seq indicated that genes involved in 7a-hydroxylation process, the first step in BA synthesis, showed pronounced alterations in skin fibroblasts or keratinocytes. The alternative pathway of BA synthesis is more actively altered than the classical pathway after ionizing radiation. In the model of rat radiogenic skin damage, DCA promoted wound healing and attenuated epidermal hyperplasia. Conclusions Ionizing radiation modulates the metabolism of BAs. DCA is a prospective therapeutic agent for the treatment of RSI.

Changes in cell wall composition due to a pectin biosynthesis enzyme GAUT10 impact root growth.

Arabidopsis (Arabidopsis thaliana) root development is regulated by multiple dynamic growth cues that require central metabolism pathways such as β-oxidation and auxin. Loss of the pectin biosynthesizing enzyme GALACTURONOSYLTRANSFERASE 10 (GAUT10) leads to a short-root phenotype under sucrose-limited conditions. The present study focused on determining the specific contributions of GAUT10 to pectin composition in primary roots and the underlying defects associated with gaut10 roots. Using live-cell microscopy, we determined reduced root growth in gaut10 is due to a reduction in both root apical meristem size and epidermal cell elongation. In addition, GAUT10 was required for normal pectin and hemicellulose composition in primary Arabidopsis roots. Specifically, loss of GAUT10 led to a reduction in galacturonic acid and xylose in root cell walls and altered the presence of rhamnogalacturonan-I (RG-I) and homogalacturonan (HG) polymers in the root. Transcriptomic analysis of gaut10 roots compared to wild type uncovered hundreds of genes differentially expressed in the mutant, including genes related to auxin metabolism and peroxisome function. Consistent with these results, both auxin signaling and metabolism were modified in gaut10 roots. The sucrose-dependent short-root phenotype in gaut10 was linked to β-oxidation based on hypersensitivity to indole-3-butyric acid (IBA) and an epistatic interaction with TRANSPORTER OF IBA1 (TOB1). Altogether, these data support a growing body of evidence suggesting that pectin composition may influence auxin pathways and peroxisome activity.

Multiple phases of yttrium-doped molybdenum trioxide nanorods as efficient dye degrader and bactericidal agents with molecular docking analysis

Contaminants removal is usually becoming an exciting subject of research from water considering their environmental and ecological effects. This work provides pathways to remove organic pollutants from water via nanomaterials and is used as an antibiotic against bacteria like Escherichia coli (E. coli). In this study, molybdenum trioxide (MoO3) and yttrium (Y) doped (2 and 4%) MoO3 nanorods were synthesized by co-precipitation method. Advanced characterization techniques have been introduced to study textural structures, morphological developments, and optical characteristics of produced products. X-ray diffraction studied multiple crystalline structures of prepared samples as hexagonal, orthorhombic, and monoclinic of pure MoO3 with decrease in crystallinity and crystallite size upon Y doping. UV–visible spectroscopy unveiled a redshift (bathochromic effect) in absorption pattern attributed to band gap energy (Eg) decreases. Photoluminescence spectra examined the recombination rate of electrons (e−) and holes (h+) as charge carriers. A sufficient catalytic activity (CA) was observed against methylene blue (MB) dye in an acidic medium (99.74%) and efficient bactericidal action was studied against (E. coli) with zone of inhibition (5.20 mm) for 4% Y-doped MoO3. In addition, in silico docking demonstrated potential inhibitory effect of produced nanomaterials on FabH and FabI enzymes of fatty acid biosynthesis.

Abscisic acid priming confers salt tolerance in maize seedlings by modulating osmotic adjustment, bond energies, ROS homeostasis, and organic acid metabolism

This study aimed at investigating the influence of exogenous abscisic acid (ABA) on salt homeostasis under 100 mM NaCl stress in maize (Zea mays L. cv. Kaveri 50) through 3 and 5 days of exposure. The ratio of Na+ to K+, hydrogen peroxide (H2O2) and superoxide (O2•‒) accumulation, electrolyte leakage were the major determinants for salt sensitivity. Pretreatment with ABA [ABA (+)] had altered the salt sensitivity of plants maximally through 5 days of treatment. Plants controlled well for endogenous ABA level (92% increase) and bond energy minimization of cell wall residues to support salt tolerance proportionately to ABA (+). Salt stress was mitigated through maintenance of relative water content (RWC) (16%), glycine betaine (GB) (26%), proline (28%) and proline biosynthesis enzyme (ΔP5CS) (26%) under the application of ABA (+). Minimization of lipid peroxides (6% decrease), carbonyl content (9% decrease), acid, alkaline phosphatase activities were more tolerated under 100 mM salinity at 5 days duration. Malate metabolism for salt tolerance was dependent on the activity of the malic enzyme, malate dehydrogenase through transcript abundance in real-time manner as a function of ABA (+). Establishment of oxidative stress through days under salinity recorded by NADPH-oxidase activity (39% increase) following ROS generation as detected in tissue specific level. The ABA (+) significantly altered redox homeostasis through ratio of AsA to DHA (21% increase), GSH to GSSG (12% increase) by dehydroascorbate reductase and glutathione reductase respectively, and other enzymes like guaiacol peroxidase, catalase, glutathione reductase activities. The ABA in priming was substantially explained in stress metabolism as biomarker for salinity stress with reference to maize.

GhXB38D represses cotton fibre elongation through ubiquitination of ethylene biosynthesis enzymes GhACS4 and GhACO1.

Ethylene plays an essential role in the development of cotton fibres. Ethylene biosynthesis in plants is elaborately regulated by the activities of key enzymes, 1-aminocyclopropane-1-carboxylate oxidase (ACO) and 1-aminocyclopropane-1-carboxylate synthase (ACS); however, the potential mechanism of post-translational modification of ACO and ACS to control ethylene synthesis in cotton fibres remains unclear. Here, we identify an E3 ubiquitin ligase, GhXB38D, that regulates ethylene biosynthesis during fibre elongation in cotton. GhXB38D gene is highly expressed in cotton fibres during the rapid elongation stage. Suppressing GhXB38D expression in cotton significantly enhanced fibre elongation and length, accompanied by the up-regulation of genes associated with ethylene signalling and fibre elongation. We demonstrated that GhXB38D interacts with the ethylene biosynthesis enzymes GhACS4 and GhACO1 in elongating fibres and specifically mediates their ubiquitination and degradation. The inhibition of GhXB38D gene expression increased the stability of GhACS4 and GhACO1 proteins in cotton fibres and ovules, resulting in an elevated concentration of ethylene. Our findings highlight the role of GhXB38D as a regulator of ethylene synthesis by ubiquitinating ACS4 and ACO1 proteins and modulating their stability. GhXB38D acts as a negative regulator of fibre elongation and serves as a potential target for enhancing cotton fibre yield and quality through gene editing strategy.

Biosynthesis of gibberellin-related compounds modulates far-red light responses in the liverwort Marchantia polymorpha.

Gibberellins (GAs) are key phytohormones that regulate growth, development, and environmental responses in angiosperms. From an evolutionary perspective, all major steps of GA biosynthesis are conserved among vascular plants, while GA biosynthesis intermediates such as ent-kaurenoic acid (KA) are also produced by bryophytes. Here, we show that in the liverwort Marchantia polymorpha, KA and GA12 are synthesized by evolutionarily conserved enzymes, which are required for developmental responses to far-red light (FR). Under FR-enriched conditions, mutants of various biosynthesis enzymes consistently exhibited altered thallus growth allometry, delayed initiation of gametogenesis, and abnormal morphology of gamete-bearing structures (gametangiophores). By chemical treatments and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses, we confirmed that these phenotypes were caused by the deficiency of some GA-related compounds derived from KA, but not bioactive GAs from vascular plants. Transcriptome analysis showed that FR enrichment induced the up-regulation of genes related to stress responses and secondary metabolism in M. polymorpha, which was largely dependent on the biosynthesis of GA-related compounds. Due to the lack of canonical GA receptors in bryophytes, we hypothesize that GA-related compounds are commonly synthesized in land plants but were co-opted independently to regulate responses to light quality change in different plant lineages during the past 450 million years of evolution.