The membrane-associated c-type cytochromes(c-Cyts) have been well known as the key enzymes mediating extracellular electron transfer to terminal electron acceptors, resulting in biogeochemical elemental transformation...The membrane-associated c-type cytochromes(c-Cyts) have been well known as the key enzymes mediating extracellular electron transfer to terminal electron acceptors, resulting in biogeochemical elemental transformation, contaminant degradation, and nutrient cycling. Although c-Cyts-mediated metal reduction or oxidation have been mainly investigated with the purified proteins of metal reducing/oxidizing bacteria, the in vivo behavior of c-Cyts is still unclear, given the difficulty in measuring the proteins of intact cells. Fortunately, the in situ spectroscopy would be ideal for measuring the reaction kinetics of c-Cyts in intact cells under noninvasive physiological conditions. It can also help the establishment of kinetic/thermodynamic models of extracellular electron transfer processes, which are essential to understand the electron transfer mechanisms at the molecular scale. This review briefly summarizes the current advances in spectral methods for examining the c-Cyts in intact cells of dissimilatory metal reducing bacteria and Fe(Ⅱ)-oxidizing bacteria.展开更多
P450 enzymes-catalyzed aromatic hydroxylation plays an important role in detoxification,biosynthesis,and potential carcinogenic effect of aromatic compounds.Though it has been explored for decades,the actual process o...P450 enzymes-catalyzed aromatic hydroxylation plays an important role in detoxification,biosynthesis,and potential carcinogenic effect of aromatic compounds.Though it has been explored for decades,the actual process of aromatic hydroxylation and mechanism of regioselectivity catalyzed by cytochrome P450 monooxygenases remained ambiguous.Here,we have resolved these issues.With a stable chiral organofluorine probe,and especially with X-ray data of two isolated arene oxides derivatives,we demonstrate that an arene oxide pathway is definitely involved in P450-catalyzed aromatic hydroxylation.By the capture,isolation,identification and reactivity exploration of the arene 1,2-oxide and arene 2,3-oxide intermediates,together with advanced QM calculations,the mechanism of how two intermediates go to the same product has been elucidated.In addition to the model substrate,we also confirmed that an arene oxide intermediate is involved in the P450-catalyzed hydroxylation pathway of a natural product derivative methyl cinnamate,which indicates that this intermediate appears to be universal in P450-catalyzed aromatic hydroxylation.Our work not only provides the most direct evidence for the arene oxide pathway and new insights into the regioselectivity involved in P450-catalyzed aromatic hydroxylation,but also supplies a new synthetic approach to achieve the dearomatization of aromatic compounds.展开更多
Rice sheath blight(RSB)is a major destructive disease impeding rice production.Identifying key germplasm resources with increased resistance remains a challenge.However,the mechanisms underlying disease resistance are...Rice sheath blight(RSB)is a major destructive disease impeding rice production.Identifying key germplasm resources with increased resistance remains a challenge.However,the mechanisms underlying disease resistance are not yet fully understood.Cytochrome P450 monooxygenases(CYP450s)serve biosynthesis and metabolic detoxification functions in plants,but there is limited information about their role in the response induced by RSB.This study demonstrated that CYT02 belongs to the CYP73A100 subfamily and is a typical member of the CYP450s.Overexpression(OE)in rice of the cytochrome P450 monooxygenase cyt02 conferred increased resistance to RSB and increased vegetative tillering.Cyt02 may increase RSB resistance by regulating plant hormone synthesis,regulate reactive oxygen species(ROS)by coordinating the activity of antioxidant enzymes,and initiate phytoalexin synthesis in response to fungal infection.These research findings have laid a foundation for a deeper understanding of the function of cyt02 and offered a potential target gene for breeding rice varieties resistant to sheath blight.展开更多
Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration ...Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Iα-loop-helix Iβmotif which corresponds to helix I of other P 450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,theγ-domain.The structure reveals a homodimer in which theγ-domain of one molecule interacts with theβ-domain of another.The plane of heme group makes an angle of approximately 40°with the helix Iα-loop-helix Iβmotif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recognition.Phe-301,Ser-303 and Gly-305 from the helix Iα-loop-helix Iβmotif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.展开更多
A class of persistent organic pollutants,polychlorinated biphenyls(PCBs),are ubiquitous in the environment and human tissues which are continuously and long-term threatened.We aim to investigate the toxic effects and ...A class of persistent organic pollutants,polychlorinated biphenyls(PCBs),are ubiquitous in the environment and human tissues which are continuously and long-term threatened.We aim to investigate the toxic effects and the underlying mechanisms of PCB153 using Caenorhabditis elegans as a model organism.Our findings demonstrated that exposure to an optimized concentration of 2μmol/L PCB153 had adverse effects on C.elegans,led to reduction of lifespan,body length,frequency of body bending,and head wiggling.Additionally,this exposure led to an increase in the accumulation of reactive oxygen species,superoxide dismutase,lipofuscin and fat content within the organism.Furthermore,gene set enrichment analysis and whole transcriptome sequencing revealed,a significant up-regulation of Cytochrome P450(CYP)family genes,which are crucial for the xenobiotics metabolism in C.elegans.Knocking down specific genes within the CYPs family,including cyp-35C1,cyp-35A5,cyp-35A3,cyp-34A1,and cyp-34A10,via RNA interference were able to reverse the shortening of lifespan and fat accumulation induced by PCB153 exposure.Moreover,a comprehensive a competing endogenous RNA network was constructed by integrating qPCR-validated differentially expressed m RNA along with lncRNA,circRNA,and miRNA identified by transcriptomic sequencing.This study emphasizes that PCB153 exposure leads to the physiological impairments,particularly fat accumulation and a shorter lifespan,through the modulation of CYP450 family genes.These findings contribute to our understanding of environmental pollutants and their impact on biological systems and provide valuable information on the toxic effects and potential risks associated with PCB153 exposure.展开更多
BACKGROUND Diabetic nephropathy(DN)is a major complication of diabetes,marked by progressive renal damage and an inflammatory response.Although research has investigated the pathological mechanisms underlying DN,effec...BACKGROUND Diabetic nephropathy(DN)is a major complication of diabetes,marked by progressive renal damage and an inflammatory response.Although research has investigated the pathological mechanisms underlying DN,effective treatment options remain limited.AIM To evaluate the therapeutic impact of Shenzhuo formulation(SZF)on a DN mouse model and to examine its potential molecular mechanisms using transcriptomic and metabolomic approaches.METHODS We established a DN mouse model through a high-fat diet combined with streptozotocin(STZ)injection,followed by SZF treatment.We analyzed SZF’s effects on gene expression and metabolite profiles in renal tissues of DN mice using transcriptomics and metabolomics techniques.Additionally,based on transcriptomic and non-targeted metabolomic findings,we further assessed SZF’s influence on the expression of factors related to the cytochrome P450(CYP450)-mediated arachidonic acid(AA)metabolism pathway,as well as its effects on inflammation and oxidative stress.RESULTS SZF intervention significantly decreased hyperglycemia and mitigated renal function impairment in DN mice.Pathological analysis revealed that SZF treatment improved renal tissue damage,reduced fibrosis,and diminished glycogen deposition.Transcriptomic analysis indicated that SZF influenced mRNA expression of CYP450-related genes,including Cyp2j13,Cyp2b9,Pla2 g2e/Cyp4a12a,Cyp4a32,Cyp2e1,and Cyp4a14.Non-targeted metabolomic results demonstrated that SZF altered the levels of metabolites associated with the AA metabolic pathway,including 5,6-EET,14,15-EET,phosphatidylcholine,and 20-HETE.Further experiments showed that SZF upregulated the expression of CYP4A and CYP2E proteins in renal tissue,as well as CYP2J and CYP2B proteins.Additionally,SZF significantly reduced the expression of inflammatory factors in renal tissue,enhanced antioxidant enzyme activity,and alleviated oxidative stress.CONCLUSION SZF exerts anti-inflammatory and antioxidant effects by regulating CYP450-mediated AA metabolism,leading to improved renal function and improved pathological state in DN mice.展开更多
The cytochrome P4503A(CYP3A)gene family’s role in early progression of gastric cancer was comprehensively investigated.Its potential as a therapeutic target was evaluated.Upon literature review,aberrant expression of...The cytochrome P4503A(CYP3A)gene family’s role in early progression of gastric cancer was comprehensively investigated.Its potential as a therapeutic target was evaluated.Upon literature review,aberrant expression of the CYP3A gene family has a strong correlation with gastric cancer onset,although the precise underlying mechanisms remain unclear.To assess its potential as a biomarker for early diagnosis and a therapeutic target,we have provided a comprehensive review of the regulatory mechanisms governing CYP3A gene family expression in gastric cancer,as well as its relation with early tumor progression and the tumor microenvironment.The CYP3A gene family is crucial in the proliferation,migration,and invasion of gastric cancer cells and promotes cancer progression by modulating inflammatory responses and oxidative stress within the tumor microenvironment.Furthermore,genetic polymorphisms in CYP3A enzymes highlight its potential value in personalized medicine.Based on these findings,this paper explores the feasibility of developing inhibitors and activators targeting CYP3A enzymes and discusses potential applications in gene therapy.This research provides crucial theoretical support for the CYP3A gene family as an early diagnostic marker and therapeutic target for gastric cancer.In the future,multi-omics studies and large-scale clinical trials will be essential to advance clinical translation of these findings.展开更多
Heme peroxygenases exhibit remarkable catalytic versatility in facilitating a wide array of oxidative reactions under mild conditions,eliminating the need for coenzymes and intricate electron transport systems.This un...Heme peroxygenases exhibit remarkable catalytic versatility in facilitating a wide array of oxidative reactions under mild conditions,eliminating the need for coenzymes and intricate electron transport systems.This unique character underscores their essentiality and potential as promising tools in synthetic biology.Recent advancements in enzyme engineering have significantly enhanced the catalytic performance of both natural and artificial peroxygenases.Extensive engineering efforts have been directed towards unspecific peroxygenases and fatty acid peroxygenases,aiming to expand their substrate specificities,and enhance reaction selectivities,as well as increase enzyme stability.Furthermore,innovative strategies such as dual-functional small molecule-assisted systems and H_(2)O_(2) tunnel engineering have been harnessed to transform P450 monooxygenases into highly efficient peroxygenases,capable of catalyzing reactions with a variety of unnatural substrates.This review consolidates the latest progress in the engineered and artificial heme peroxygenases,emphasizing their catalytic performances as potent biocatalysts for sustainable organic synthesis.展开更多
The tobacco whitefly,Bemisia tabaci,is a notorious pest affecting various crops globally,and it exhibits high levels of resistance to various insecticides.Afidopyropen is a recently commercialized pyropene insecticide...The tobacco whitefly,Bemisia tabaci,is a notorious pest affecting various crops globally,and it exhibits high levels of resistance to various insecticides.Afidopyropen is a recently commercialized pyropene insecticide for B.tabaci control with high selectivity and a novel mode of action.We previously identified a high level of afidopyropen resistance in a field-collected population after selection in the lab,and named it the HD-Afi strain.In the present study,minimal cross-resistance in the HD-Afi strain was found between afidopyropen and other common chemical agents.However,the P450 enzyme activity in HD-Afi was 2.18 times the level in susceptible strain HD-S.Expression analysis revealed that two of 12 candidate P450 genes,namely CYP6DW3 and CYP4C64,were significantly up-regulated in HD-Afi.Silencing CYP6DW3 and CYP4C64 by RNA interference(RNAi)substantially increased the susceptibility of whitefly adults,confirming their involvement in afidopyropen resistance.Homology modeling and molecular docking analyses demonstrated stable binding of afidopyropen to CYP6DW3 and CYP4C64,with binding free energies of–6.87 and–6.11 kcal mol^(-1),respectively.The findings of this study suggest that the induction of CYP6DW3 and CYP4C64 facilitates afidopyropen detoxification,contributing to the development of resistance in B.tabaci.展开更多
Background:The thermogenic characteristics of animals are closely related to species distribution,and basal metabolic rate(BMR)and thermal neutral zone(TNZ)are important components of the thermogenic process in animal...Background:The thermogenic characteristics of animals are closely related to species distribution,and basal metabolic rate(BMR)and thermal neutral zone(TNZ)are important components of the thermogenic process in animals.Furthermore,the cytochrome c oxidase 1(COX1)gene has become a subject of particular interest due to its high degree of sequence conservation,stable evolutionary rate,and rare insertions/deletions.Method:The present study selected 29 species of Rodentia and 20 species of Chiroptera.The present study employed the statistical software SPSS(27.0 Chinese version)and Origin(2024 Chinese version)software to conduct correlation analyses on a variety of biological and ecological variables.These variables included body weight,BMR,TNZ,upper thermal neutral zone,lower thermal neutral zone(LTNZ),litter size,and dietary patterns of species from different latitudes.Furthermore,we conducted a series of phylogenetic tree analyses on COX1 protein.Results:As the geographic location of a species increases in latitude,there is an observed upward trend in both body mass and BMR of rodent species.Rodentia and Chiroptera species have been observed to exhibit a decrease in LTNZ and an expansion of TNZ.With regard to dietary habits,Rodentia species are predominantly phytophagous or omnivorous.Omnivorous species exhibit a marked tendency to produce larger litters in comparison to their herbivorous counterparts.Chiroptera species exhibit a diverse dietary range,including phytophagous,carnivorous,and omnivorous species.However,no correlation was observed between dietary differences and litter size.Phylogenetic analysis of the COX1 protein subsequently demonstrated that these two species groups share a monophyletic origin.Conclusion:The present study suggests that the selected Rodentia and Chiroptera species adapt to high-latitude environments by lowering the LTNZ and widening the TNZ.Furthermore,an upward trend has been observed in the body mass and BMR of high-latitude Rodentia.Phylogenetic analysis of the COX1 protein across various taxonomic groups substantiates the efficacy of this gene for species identification.Integrating physiological phenotypes with COX1 protein molecular evidence,this study provides a reference framework for the multidimensional mechanisms of mammalian latitudinal adaptation.展开更多
The high content of cyanogenic glycosides(CG)in cassava tubers affects food safety.CG are involved in the plant growth and development and protect cassava leaves from herbivorous predators.However,the regulatory mecha...The high content of cyanogenic glycosides(CG)in cassava tubers affects food safety.CG are involved in the plant growth and development and protect cassava leaves from herbivorous predators.However,the regulatory mechanism of CG biosynthesis remains poorly understood.Here,yeast one-hybrid assays were performed using a mixed cDNA library of cassava tubers and leaves as prey and the promoter of MeCYP79D2 as bait.MeCYP79D2,a cytochrome P450 protein,is the rate-limiting enzyme for CG synthesis in cassava.From this information,a candidate regulator of MeCYP79D2 was selected and identified as transcription factor MePHD1.2.MePHD1.2,located in the nucleus and exhibiting an inhibitory transcription activity directly bound to an AT-rich motif in the promoter of MeCYP79D2.In cassava,the transcriptional activity of MeCYP79D2 was considerably enhanced in mephd1.2 mutant lines leading to increased linamarin and lotaustralin contents.Deletion of MePHD1.2 promoted the production of CGs in cassava and decreased transcription inhibition on MeCYP79D2,exposing a novel regulatory module governing biosynthesis of CGs.展开更多
Benzylisoquinoline alkaloids(BIAs)are a structurally diverse group of plant metabolites renowned for their pharmacological properties.However,sustainable sources for these compounds remain limited.Consequently,researc...Benzylisoquinoline alkaloids(BIAs)are a structurally diverse group of plant metabolites renowned for their pharmacological properties.However,sustainable sources for these compounds remain limited.Consequently,researchers are focusing on elucidating BIA biosynthetic pathways and genes to explore alternative sources using synthetic biology approaches.CYP80B,a family of cytochrome P450(CYP450)enzymes,plays a crucial role in BIA biosynthesis.Previously reported CYP80Bs are known to catalyze the 3′-hydroxylation of(S)-Nmethylcoclaurine,with the N-methyl group essential for catalytic activity.In this study,we successfully cloned a full-length CYP80B gene(St CYP80B)from Stephania tetrandra(S.tetrandra)and identified its function using a yeast heterologous expression system.Both in vivo yeast feeding and in vitro enzyme analysis demonstrated that St CYP80B could catalyze Nmethylcoclaurine and coclaurine into their respective 3'-hydroxylated products.Notably,St CYP80B exhibited an expanded substrate selectivity compared to previously reported wildtype CYP80Bs,as it did not require an N-methyl group for hydroxylase activity.Furthermore,St CYP80B displayed a clear preference for the(S)-configuration.Co-expression of St CYP80B with the CYP450 reductases(CPRs,StCPR1,and StCPR2),also cloned from S.tetrandra,significantly enhanced the catalytic activity towards(S)-coclaurine.Site-directed mutagenesis of St CYP80B revealed that the residue H205 is crucial for coclaurine catalysis.Additionally,St CYP80B exhibited tissue-specific expression in plants.This study provides new genetic resources for the biosynthesis of BIAs and further elucidates their synthetic pathway in natural plant systems.展开更多
Understanding the genetic diversity–area relationship(GAR)is essential for comprehending how species adapt to environmental changes,as genetic diversity is an indicator of a species’adaptive potential.Variation in e...Understanding the genetic diversity–area relationship(GAR)is essential for comprehending how species adapt to environmental changes,as genetic diversity is an indicator of a species’adaptive potential.Variation in environmental adaptation capacity exists among species and animal taxa with different distribution areas,highlighting the importance of understanding the GAR.To obtain a more comprehensive understanding of the GAR in terrestrial vertebrates,we assessed both haplotype diversity–area and nucleotide diversity–area relationships using 25,453 cytochrome c oxidase subunit I(COI)sequences from 142 amphibian species,574 bird species,and 342 mammal species.We found that both measures of genetic diversity increased with species range size across major animal groups.Nevertheless,the GAR did not differ among animal groups,while haplotype diversity performed better than nucleotide diversity in profiling the GAR,as indicated by higher R2 values.The difference in the modeling fit may stem from the distinct biological and mathematical significance of nucleotide diversity and haplotype diversity.These results suggest that the GAR follows similar rules among different animal taxa.Furthermore,haplotype diversity may serve as a more reliable indicator for assessing the potential effects of area size changes on animal populations and provide better guidance for conserving genetic diversity.展开更多
High molecular weight polycyclic aromatic hydrocarbons(HMW-PAHs)pose significant environmental challenges due to their complex structures and persistent toxicity,and are difficult to be degraded by bacteria.Fungi,howe...High molecular weight polycyclic aromatic hydrocarbons(HMW-PAHs)pose significant environmental challenges due to their complex structures and persistent toxicity,and are difficult to be degraded by bacteria.Fungi,however,possess the ability to overcome these challenges,primarily through various enzymes with broad substrate specificity,including cytochrome P450(CYP450),laccase,manganese peroxidase,lignin peroxidase.As a result,the development of efficient fungal strains capable of degrading HMW-PAHs is essential for advancing bioremediation strategies.In this study,Aspergillus fumigatus Z5 was isolated from a contaminated site and demonstrated a remarkable ability to degrade the highly recalcitrant benzo[a]pyrene,achieving a degradation rate of 75.43%.Transcriptomic analysis revealed significant upregulation of 34 CYP450 genes.Among these genes,CYP3A4(gene 8840)showed strong binding affinity for benzo[a]pyrene,as confirmed by molecular docking studies,indicating its key role in the biodegradable process.Furthermore,the metabolic network analysis showed that the electron transfer required for CYP450-mediated oxidative reactions enhanced mitochondrial oxidative phosphorylation and subsequently energy metabolism in A.fumigatus Z5.This metabolic coordination likely facilitates the complete degradation of benzo[a]pyrene.Our study illuminates the fundamental roles of CYP450 from A.fumigatus Z5 in benzo[a]pyrene degradation and provides novel insight into designing and implementing enhanced bioremediation strategies.展开更多
Background:As a major histopathological subtype of gastric cancer(GC),stomach adenocarcinoma(STAD)is an important malignant tumor in the digestive system.Increasing evidence also indicates that endoplasmic reticulum(E...Background:As a major histopathological subtype of gastric cancer(GC),stomach adenocarcinoma(STAD)is an important malignant tumor in the digestive system.Increasing evidence also indicates that endoplasmic reticulum(ER)stress plays a pivotal role in the pathogenesis and progression of GC.Therefore,this study aims to screen and identify vital ER stress-related genes that could contribute to the malignant development and poor prognosis for STAD.Methods:A novel ER stress-related risk score signature was developed employingmachine learning techniques.Then,a prognostic prediction nomogram was also built based on the clinicopathological characteristics and the risk score signature.The tumor immune microenvironment characteristics and pathway enrichment analysis in different risk groups were also explored.Furthermore,through the single-cell RNA sequencing(scRNA-seq)analysis,the study highlightedCytochrome P450 Family 19 SubfamilyAMember 1(CYP19A1)as the pivotal research target and detected its effect on cell proliferation by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide(MTT)and the expression of ER stress-related genes by RT-qPCR in STAD.Results:Based on the evaluation of five screened key ER stressrelated genes(AKR1B1,SERPINE1,ADCYAP1,MATN3,CYP19A1),our ER stress-related risk score signature offers a novel approach for assessing STAD prognosis hazards.The novel prognostic prediction nomogram based on the signature also accurately predicted the survival outcomes of patients with STAD.Furthermore,the expression of CYP19A1 is significantly higher in STAD tissues than in normal tissues.High expression of CYP19A1 was related to a poor survival outcome for patients with STAD.Besides,compared to normal gastric epithelial cells,the expression of CYP19A1 was significantly higher in STAD cell lines.Silencing the expression of CYP19A1 significantly inhibited the cell proliferation ability and decreased the expression of ER stress-related genes,including ATF4,DDIT3 and XBP1 in STAD.Conclusions:In conclusion,our study developed a novel prognosis prediction signature and identified the novel diagnostic and therapeutic target CYP19A1 for patients with STAD.展开更多
Background:Hepatocellular carcinoma(HCC)is a prevalent liver malignancy.This study examined the roles of transforming growth factor beta(TGF-β)and cytochrome b5 domain containing 2(CYB5D2)in HCC etiology and their pr...Background:Hepatocellular carcinoma(HCC)is a prevalent liver malignancy.This study examined the roles of transforming growth factor beta(TGF-β)and cytochrome b5 domain containing 2(CYB5D2)in HCC etiology and their prognostic biomarker potential.Methods:Key modules and prognostic genes were identified by analyzing the GSE101685 dataset by weighted gene co-expression network analysis(WGCNA)and Least absolute shrinkage and selection operator(LASSO)Cox regression.The expression levels of CYB5D2 and TGF-βin HCC cell lines were quantified using Quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western blotting(WB)assays.Effects of CYB5D2 overexpression on cell proliferation,migration,invasion,and epithelial-mesenchymal transition(EMT)marker regulation were assessed in vitro,while in vivo tumorigenicity was evaluated using a xenograft model of HCC in nude mice.Results:In this study,WGCNA identified the turquoise module as significantly associated with HCC,containing 452 DEGs.LASSO Cox regression analysis revealed 9 key prognostic genes,with CYB5D2 being underexpressed in HCC cells and tissues.TGF-βwas negatively correlated with CYB5D2 expression,resulting in poor patient prognosis.Functional assays demonstrated that CYB5D2 overexpression inhibited proliferation,migration,and invasion of HCC cell lines,and altered EMT marker expression.Furthermore,the addition of TGF-βpartially reversed the suppressive effects caused by CYB5D2 overexpression.In vivo,CYB5D2 overexpression significantly reduced tumor growth,indicating its potential as a therapeutic target for HCC.Conclusion:The tumor suppressor function of CYB5D2 in HCC and its interaction with TGF-βoffered fresh information on the molecular pathophysiology of HCC and possible treatment avenues.展开更多
BACKGROUND Kidney transplantation is an effective renal replacement therapy for improving survival and quality of life in chronic kidney disease patients.Kidney transplant recipients need lifelong immunosuppression to...BACKGROUND Kidney transplantation is an effective renal replacement therapy for improving survival and quality of life in chronic kidney disease patients.Kidney transplant recipients need lifelong immunosuppression to prevent rejection and allograft dysfunction.Tacrolimus,a calcineurin inhibitor,is metabolized differently based on cytochrome P4503A(CYP3A)5 genetic variations and this impacts the graft outcome.AIM To examine the clinical outcomes in kidney transplant recipients affected by the variable metabolism of tacrolimus due to the CYP3A5 genetic variation,emphasizing personalized immunosuppression strategies to optimize efficacy,minimize toxicity,and enhance long-term graft survival.METHODS A retrospective study was conducted at a tertiary care center in Central India on 95 kidney transplant recipients.Patient demographics,medical history,CYP3A5 polymorphism,post-transplant investigations,graft biopsy results,preexisting comorbidities,history of post–kidney transplant infections,and new onset diabetes after transplantation(NODAT)was collected.Tacrolimus was initiated at 0.1 mg/kg/day for CYP3A5 expressors and 0.05 mg/kg/day for non-expressors,with dose adjustments to maintain target C0 levels of 7-10 ng/mL for first 6 months and 5-7 ng/mL from 6 months to 12 months posttransplant.Patients were followed regularly for one year for glomerular filtration rate(GFR),creatinine,and the tacrolimus trough concentration(ng/mL)/daily tacrolimus dose(mg/kg/day)ratio(C/D).A P value≤0.05 was considered statistically significant.RESULTS Kidney transplant recipients were classified as expressors(CYP3A51 carriers,n=35)and non-expressors(CYP3A5*3*3,n=60).Both groups were comparable for age,sex,and donor characteristics.Tacrolimus dose was comparable post-transplant except at 6 months and 12 months,where expressors required higher doses.Kidney function(creatinine and estimated GFR),NODAT,hypomagnesemia,and infections showed no significant differences between the two groups over 12 months of follow-up.Biopsy-proven acute rejection(BPAR)was found to be more in expressors(22.9%vs 13.3%,P=0.2340)though it was not found to be statistically significant.Nonexpressors had a significantly higher tacrolimus levels and C/D ratio at multiple follow-ups.CONCLUSION CYP3A5 expressors require higher tacrolimus doses to maintain therapeutic levels as compared to non-expressors.BPAR was higher in expressors but the difference was not significant.Graft function,infection rate,and NODAT were comparable irrespective of CYP3A5 expression status,emphasizing the importance of pretransplant CYP3A5 genotyping and therapeutic drug monitoring to guide tacrolimus dosing for individualized immunosuppressive management.展开更多
D-a-Tocopheryl polyethylene glycol 1000 succinate(TPGS,also known as vitamin E-TPGS)is a biodegradable amphiphilic polymer prepared by esterification of vitamin E with polyethylene glycol(PEG)1000.It is approved by th...D-a-Tocopheryl polyethylene glycol 1000 succinate(TPGS,also known as vitamin E-TPGS)is a biodegradable amphiphilic polymer prepared by esterification of vitamin E with polyethylene glycol(PEG)1000.It is approved by the US Food and Drug Administration(FDA)and has found wide application in nanocarrier drug delivery systems(NDDS).Fully characterizing the in vivo fate and pharmacokinetic behavior of TPGS is important to promote the further development of TPGS-based NDDS.However,to date,a bioassay for the simultaneous quantitation of TPGS and its metabolite,PEG1000,has not been reported.In the present study,we developed such an innovative bioassay and used it to investigate the pharmacokinetics,tissue distribution and excretion of TPGS and PEG1000 in rat after oral and intravenous dosing.In addition,we evaluated the interaction of TPGS with cytochromes P450(CYP450s)in human liver microsomes.The results show that TPGS is poorly absorbed after oral administration with very low bioavailability and that,after intravenous administration,TPGS and PEG1000 are mainly distributed to the spleen,liver,lung and kidney before both being slowly eliminated in urine and feces as PEG1000.In vitro studies show the inhibition of human CYP450 enzymes by TPGS is limited to a weak inhibition of CYP3A4.Overall,our results provide a clear picture of the in vivo fate of TPGS which will be useful in evaluating the safety of TPGS-based NDDS in clinical use and in promoting their further development.展开更多
Direct interspecies electron transfer(DIET)may be most important in methanogenic environments,but mechanistic studies of DIET to date have primarily focused on cocultures in which fumarate was the terminal electron ac...Direct interspecies electron transfer(DIET)may be most important in methanogenic environments,but mechanistic studies of DIET to date have primarily focused on cocultures in which fumarate was the terminal electron acceptor.To better understand DIET with methanogens,the transcriptome of Geobacter metallireducens during DIET‐based growth with G.sulfurreducens reducing fumarate was compared with G.metallireducens grown in coculture with diverse Methanosarcina.The transcriptome of G.metallireducens cocultured with G.sulfurreducens was significantly different from those with Methanosarcina.Furthermore,the transcriptome of G.metallireducens grown with Methanosarcina barkeri,which lacks outer‐surface c‐type cytochromes,differed from those of G.metallireducens cocultured with M.acetivorans or M.subterranea,which have an outer‐surface c‐type cytochrome that serves as an electrical connect for DIET.Differences in G.metallireducens expression patterns for genes involved in extracellular electron transfer were particularly notable.Cocultures with c‐type cytochrome deletion mutant strains,ΔGmet_0930,ΔGmet_0557 andΔGmet_2896,never became established with G.sulfurreducens but adapted to grow with all three Methanosarcina.Two porin–cytochrome complexes,PccF and PccG,were important for DIET;however,PccG was more important for growth with Methanosarcina.Unlike cocultures with G.sulfurreducens and M.acetivorans,electrically conductive pili were not needed for growth with M.barkeri.Shewanella oneidensis,another electroactive microbe with abundant outer‐surface c‐type cytochromes,did not grow via DIET.The results demonstrate that the presence of outer‐surface c‐type cytochromes does not necessarily confer the capacity for DIET and emphasize the impact of the electron‐accepting partner on the physiology of the electron‐donating DIET partner.展开更多
Extracellular electron transfer(EET)plays a critical role in bioelectrochemical processes,allowing cou-pling between microorganisms and extracellular solid-state electrodes,metals,or other cells in energy metabolism.P...Extracellular electron transfer(EET)plays a critical role in bioelectrochemical processes,allowing cou-pling between microorganisms and extracellular solid-state electrodes,metals,or other cells in energy metabolism.Previous studies have suggested a role for outer-surface c-type cytochromes in direct metal-to-microbe electron transfer by Geobacter sulfurreducens,a model electroactive bacterium.Here,we ex-amined the possibility of other microbially produced electrical contacts by deleting the gene for PilA,the protein monomer that G.sulfurreducens assembles into electrically conductive protein nanowires(e-pili).Deleting pilA gene inhibited electron extraction from pure iron and 316L stainless steel up to 31%and 81%,respectively more than deleting the gene for the outer-surface cytochrome OmcS.This PilA-deficient phenotype,and the observation that relatively thick biofilms(21.7μm)grew on the metal surfaces at multi-cell distances from the metal surfaces suggest that e-pili contributed significantly to microbial cor-rosion via direct metal-to-microbe electron transfer.These results have implications for the fundamental understanding of electron harvest via e-pili by electroactive microbes,their uses in bioenergy production,as well as in monitoring and mitigation of metal biocorrosion.展开更多
基金funded by the National Natural Science Foundations of China(41522105 and 41571130052)Guangdong Natural Science Funds for Distinguished Young Scholar(2014A030306041)Special Support Program(2016)
文摘The membrane-associated c-type cytochromes(c-Cyts) have been well known as the key enzymes mediating extracellular electron transfer to terminal electron acceptors, resulting in biogeochemical elemental transformation, contaminant degradation, and nutrient cycling. Although c-Cyts-mediated metal reduction or oxidation have been mainly investigated with the purified proteins of metal reducing/oxidizing bacteria, the in vivo behavior of c-Cyts is still unclear, given the difficulty in measuring the proteins of intact cells. Fortunately, the in situ spectroscopy would be ideal for measuring the reaction kinetics of c-Cyts in intact cells under noninvasive physiological conditions. It can also help the establishment of kinetic/thermodynamic models of extracellular electron transfer processes, which are essential to understand the electron transfer mechanisms at the molecular scale. This review briefly summarizes the current advances in spectral methods for examining the c-Cyts in intact cells of dissimilatory metal reducing bacteria and Fe(Ⅱ)-oxidizing bacteria.
文摘P450 enzymes-catalyzed aromatic hydroxylation plays an important role in detoxification,biosynthesis,and potential carcinogenic effect of aromatic compounds.Though it has been explored for decades,the actual process of aromatic hydroxylation and mechanism of regioselectivity catalyzed by cytochrome P450 monooxygenases remained ambiguous.Here,we have resolved these issues.With a stable chiral organofluorine probe,and especially with X-ray data of two isolated arene oxides derivatives,we demonstrate that an arene oxide pathway is definitely involved in P450-catalyzed aromatic hydroxylation.By the capture,isolation,identification and reactivity exploration of the arene 1,2-oxide and arene 2,3-oxide intermediates,together with advanced QM calculations,the mechanism of how two intermediates go to the same product has been elucidated.In addition to the model substrate,we also confirmed that an arene oxide intermediate is involved in the P450-catalyzed hydroxylation pathway of a natural product derivative methyl cinnamate,which indicates that this intermediate appears to be universal in P450-catalyzed aromatic hydroxylation.Our work not only provides the most direct evidence for the arene oxide pathway and new insights into the regioselectivity involved in P450-catalyzed aromatic hydroxylation,but also supplies a new synthetic approach to achieve the dearomatization of aromatic compounds.
基金supported by the Sichuan Province International Science and Technology Innovation Cooperation(2024YFHZ0299)the Project of Science and Technology Department of Sichuan Province(2022YFH0031)Chengdu Science and Technology Bureau(2024-YF05-02168-SN).
文摘Rice sheath blight(RSB)is a major destructive disease impeding rice production.Identifying key germplasm resources with increased resistance remains a challenge.However,the mechanisms underlying disease resistance are not yet fully understood.Cytochrome P450 monooxygenases(CYP450s)serve biosynthesis and metabolic detoxification functions in plants,but there is limited information about their role in the response induced by RSB.This study demonstrated that CYT02 belongs to the CYP73A100 subfamily and is a typical member of the CYP450s.Overexpression(OE)in rice of the cytochrome P450 monooxygenase cyt02 conferred increased resistance to RSB and increased vegetative tillering.Cyt02 may increase RSB resistance by regulating plant hormone synthesis,regulate reactive oxygen species(ROS)by coordinating the activity of antioxidant enzymes,and initiate phytoalexin synthesis in response to fungal infection.These research findings have laid a foundation for a deeper understanding of the function of cyt02 and offered a potential target gene for breeding rice varieties resistant to sheath blight.
文摘Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Iα-loop-helix Iβmotif which corresponds to helix I of other P 450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,theγ-domain.The structure reveals a homodimer in which theγ-domain of one molecule interacts with theβ-domain of another.The plane of heme group makes an angle of approximately 40°with the helix Iα-loop-helix Iβmotif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recognition.Phe-301,Ser-303 and Gly-305 from the helix Iα-loop-helix Iβmotif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.
基金supported by the National Natural Science Foundation of China(No.81872584)the Natural Science Foundation of Shenzhen(No.JCYJ20210324093211030)+1 种基金Henan Province’s Key R&D and Promotion Projects(No.252102310075)the Interdisciplinary Research for Firstclass Discipline Construction Project of Henan University(No.2019YLXKJC04)。
文摘A class of persistent organic pollutants,polychlorinated biphenyls(PCBs),are ubiquitous in the environment and human tissues which are continuously and long-term threatened.We aim to investigate the toxic effects and the underlying mechanisms of PCB153 using Caenorhabditis elegans as a model organism.Our findings demonstrated that exposure to an optimized concentration of 2μmol/L PCB153 had adverse effects on C.elegans,led to reduction of lifespan,body length,frequency of body bending,and head wiggling.Additionally,this exposure led to an increase in the accumulation of reactive oxygen species,superoxide dismutase,lipofuscin and fat content within the organism.Furthermore,gene set enrichment analysis and whole transcriptome sequencing revealed,a significant up-regulation of Cytochrome P450(CYP)family genes,which are crucial for the xenobiotics metabolism in C.elegans.Knocking down specific genes within the CYPs family,including cyp-35C1,cyp-35A5,cyp-35A3,cyp-34A1,and cyp-34A10,via RNA interference were able to reverse the shortening of lifespan and fat accumulation induced by PCB153 exposure.Moreover,a comprehensive a competing endogenous RNA network was constructed by integrating qPCR-validated differentially expressed m RNA along with lncRNA,circRNA,and miRNA identified by transcriptomic sequencing.This study emphasizes that PCB153 exposure leads to the physiological impairments,particularly fat accumulation and a shorter lifespan,through the modulation of CYP450 family genes.These findings contribute to our understanding of environmental pollutants and their impact on biological systems and provide valuable information on the toxic effects and potential risks associated with PCB153 exposure.
基金Supported by the Natural Science Foundation of Hebei Province Beijing-Tianjin-Hebei Basic Research Special Program,No.H2020110287Key Laboratory for Diabetic Kidney Disease Syndrome and Treatment of the Traditional Chinese Medicine Administration of Hebei Province,No.7[2024]+2 种基金Yuansong Wang National Famous Traditional Chinese Medicine Expert Heritage Studio,No.3[2024]Hebei Key Laboratory of Integrated Traditional Chinese and Western Medicine for Diabetes and Its Complications,No.SZX2020015Jiangsu Province Chinese Medicine Science and Technology Development Program Project,No.YB2020065.
文摘BACKGROUND Diabetic nephropathy(DN)is a major complication of diabetes,marked by progressive renal damage and an inflammatory response.Although research has investigated the pathological mechanisms underlying DN,effective treatment options remain limited.AIM To evaluate the therapeutic impact of Shenzhuo formulation(SZF)on a DN mouse model and to examine its potential molecular mechanisms using transcriptomic and metabolomic approaches.METHODS We established a DN mouse model through a high-fat diet combined with streptozotocin(STZ)injection,followed by SZF treatment.We analyzed SZF’s effects on gene expression and metabolite profiles in renal tissues of DN mice using transcriptomics and metabolomics techniques.Additionally,based on transcriptomic and non-targeted metabolomic findings,we further assessed SZF’s influence on the expression of factors related to the cytochrome P450(CYP450)-mediated arachidonic acid(AA)metabolism pathway,as well as its effects on inflammation and oxidative stress.RESULTS SZF intervention significantly decreased hyperglycemia and mitigated renal function impairment in DN mice.Pathological analysis revealed that SZF treatment improved renal tissue damage,reduced fibrosis,and diminished glycogen deposition.Transcriptomic analysis indicated that SZF influenced mRNA expression of CYP450-related genes,including Cyp2j13,Cyp2b9,Pla2 g2e/Cyp4a12a,Cyp4a32,Cyp2e1,and Cyp4a14.Non-targeted metabolomic results demonstrated that SZF altered the levels of metabolites associated with the AA metabolic pathway,including 5,6-EET,14,15-EET,phosphatidylcholine,and 20-HETE.Further experiments showed that SZF upregulated the expression of CYP4A and CYP2E proteins in renal tissue,as well as CYP2J and CYP2B proteins.Additionally,SZF significantly reduced the expression of inflammatory factors in renal tissue,enhanced antioxidant enzyme activity,and alleviated oxidative stress.CONCLUSION SZF exerts anti-inflammatory and antioxidant effects by regulating CYP450-mediated AA metabolism,leading to improved renal function and improved pathological state in DN mice.
文摘The cytochrome P4503A(CYP3A)gene family’s role in early progression of gastric cancer was comprehensively investigated.Its potential as a therapeutic target was evaluated.Upon literature review,aberrant expression of the CYP3A gene family has a strong correlation with gastric cancer onset,although the precise underlying mechanisms remain unclear.To assess its potential as a biomarker for early diagnosis and a therapeutic target,we have provided a comprehensive review of the regulatory mechanisms governing CYP3A gene family expression in gastric cancer,as well as its relation with early tumor progression and the tumor microenvironment.The CYP3A gene family is crucial in the proliferation,migration,and invasion of gastric cancer cells and promotes cancer progression by modulating inflammatory responses and oxidative stress within the tumor microenvironment.Furthermore,genetic polymorphisms in CYP3A enzymes highlight its potential value in personalized medicine.Based on these findings,this paper explores the feasibility of developing inhibitors and activators targeting CYP3A enzymes and discusses potential applications in gene therapy.This research provides crucial theoretical support for the CYP3A gene family as an early diagnostic marker and therapeutic target for gastric cancer.In the future,multi-omics studies and large-scale clinical trials will be essential to advance clinical translation of these findings.
文摘Heme peroxygenases exhibit remarkable catalytic versatility in facilitating a wide array of oxidative reactions under mild conditions,eliminating the need for coenzymes and intricate electron transport systems.This unique character underscores their essentiality and potential as promising tools in synthetic biology.Recent advancements in enzyme engineering have significantly enhanced the catalytic performance of both natural and artificial peroxygenases.Extensive engineering efforts have been directed towards unspecific peroxygenases and fatty acid peroxygenases,aiming to expand their substrate specificities,and enhance reaction selectivities,as well as increase enzyme stability.Furthermore,innovative strategies such as dual-functional small molecule-assisted systems and H_(2)O_(2) tunnel engineering have been harnessed to transform P450 monooxygenases into highly efficient peroxygenases,capable of catalyzing reactions with a variety of unnatural substrates.This review consolidates the latest progress in the engineered and artificial heme peroxygenases,emphasizing their catalytic performances as potent biocatalysts for sustainable organic synthesis.
基金partly supported by research grants from the Outstanding Youth Foundation of the Beijing Academy of Agriculture and Forestry Sciences,China(YXQN202301)the National Natural Science Foundation of China(32272522)the Beijing Natural Science Foundation,China(6232005)。
文摘The tobacco whitefly,Bemisia tabaci,is a notorious pest affecting various crops globally,and it exhibits high levels of resistance to various insecticides.Afidopyropen is a recently commercialized pyropene insecticide for B.tabaci control with high selectivity and a novel mode of action.We previously identified a high level of afidopyropen resistance in a field-collected population after selection in the lab,and named it the HD-Afi strain.In the present study,minimal cross-resistance in the HD-Afi strain was found between afidopyropen and other common chemical agents.However,the P450 enzyme activity in HD-Afi was 2.18 times the level in susceptible strain HD-S.Expression analysis revealed that two of 12 candidate P450 genes,namely CYP6DW3 and CYP4C64,were significantly up-regulated in HD-Afi.Silencing CYP6DW3 and CYP4C64 by RNA interference(RNAi)substantially increased the susceptibility of whitefly adults,confirming their involvement in afidopyropen resistance.Homology modeling and molecular docking analyses demonstrated stable binding of afidopyropen to CYP6DW3 and CYP4C64,with binding free energies of–6.87 and–6.11 kcal mol^(-1),respectively.The findings of this study suggest that the induction of CYP6DW3 and CYP4C64 facilitates afidopyropen detoxification,contributing to the development of resistance in B.tabaci.
基金supported by the National Natural Scientific Foundation of China(32160254)Yunnan Fundamental Research Projects(202401AS070039).
文摘Background:The thermogenic characteristics of animals are closely related to species distribution,and basal metabolic rate(BMR)and thermal neutral zone(TNZ)are important components of the thermogenic process in animals.Furthermore,the cytochrome c oxidase 1(COX1)gene has become a subject of particular interest due to its high degree of sequence conservation,stable evolutionary rate,and rare insertions/deletions.Method:The present study selected 29 species of Rodentia and 20 species of Chiroptera.The present study employed the statistical software SPSS(27.0 Chinese version)and Origin(2024 Chinese version)software to conduct correlation analyses on a variety of biological and ecological variables.These variables included body weight,BMR,TNZ,upper thermal neutral zone,lower thermal neutral zone(LTNZ),litter size,and dietary patterns of species from different latitudes.Furthermore,we conducted a series of phylogenetic tree analyses on COX1 protein.Results:As the geographic location of a species increases in latitude,there is an observed upward trend in both body mass and BMR of rodent species.Rodentia and Chiroptera species have been observed to exhibit a decrease in LTNZ and an expansion of TNZ.With regard to dietary habits,Rodentia species are predominantly phytophagous or omnivorous.Omnivorous species exhibit a marked tendency to produce larger litters in comparison to their herbivorous counterparts.Chiroptera species exhibit a diverse dietary range,including phytophagous,carnivorous,and omnivorous species.However,no correlation was observed between dietary differences and litter size.Phylogenetic analysis of the COX1 protein subsequently demonstrated that these two species groups share a monophyletic origin.Conclusion:The present study suggests that the selected Rodentia and Chiroptera species adapt to high-latitude environments by lowering the LTNZ and widening the TNZ.Furthermore,an upward trend has been observed in the body mass and BMR of high-latitude Rodentia.Phylogenetic analysis of the COX1 protein across various taxonomic groups substantiates the efficacy of this gene for species identification.Integrating physiological phenotypes with COX1 protein molecular evidence,this study provides a reference framework for the multidimensional mechanisms of mammalian latitudinal adaptation.
基金supported by grants from the National Natural Science Foundation of China(32460505)China Agriculture Research System(CARS-11)+2 种基金the Chinese Academy of Tropical Agricultural Sciences for the Science and Technology Innovation Team of the National Tropical Agricultural Science Center(CATASCXTD202301)Additional support was provided by the Hainan Province Graduate Innovation Research Project(Hyb2020-09)the Key Laboratory of Tropical Fruits and Vegetables Quality and Safety for State Market Regulation(KF-2023016).
文摘The high content of cyanogenic glycosides(CG)in cassava tubers affects food safety.CG are involved in the plant growth and development and protect cassava leaves from herbivorous predators.However,the regulatory mechanism of CG biosynthesis remains poorly understood.Here,yeast one-hybrid assays were performed using a mixed cDNA library of cassava tubers and leaves as prey and the promoter of MeCYP79D2 as bait.MeCYP79D2,a cytochrome P450 protein,is the rate-limiting enzyme for CG synthesis in cassava.From this information,a candidate regulator of MeCYP79D2 was selected and identified as transcription factor MePHD1.2.MePHD1.2,located in the nucleus and exhibiting an inhibitory transcription activity directly bound to an AT-rich motif in the promoter of MeCYP79D2.In cassava,the transcriptional activity of MeCYP79D2 was considerably enhanced in mephd1.2 mutant lines leading to increased linamarin and lotaustralin contents.Deletion of MePHD1.2 promoted the production of CGs in cassava and decreased transcription inhibition on MeCYP79D2,exposing a novel regulatory module governing biosynthesis of CGs.
文摘Benzylisoquinoline alkaloids(BIAs)are a structurally diverse group of plant metabolites renowned for their pharmacological properties.However,sustainable sources for these compounds remain limited.Consequently,researchers are focusing on elucidating BIA biosynthetic pathways and genes to explore alternative sources using synthetic biology approaches.CYP80B,a family of cytochrome P450(CYP450)enzymes,plays a crucial role in BIA biosynthesis.Previously reported CYP80Bs are known to catalyze the 3′-hydroxylation of(S)-Nmethylcoclaurine,with the N-methyl group essential for catalytic activity.In this study,we successfully cloned a full-length CYP80B gene(St CYP80B)from Stephania tetrandra(S.tetrandra)and identified its function using a yeast heterologous expression system.Both in vivo yeast feeding and in vitro enzyme analysis demonstrated that St CYP80B could catalyze Nmethylcoclaurine and coclaurine into their respective 3'-hydroxylated products.Notably,St CYP80B exhibited an expanded substrate selectivity compared to previously reported wildtype CYP80Bs,as it did not require an N-methyl group for hydroxylase activity.Furthermore,St CYP80B displayed a clear preference for the(S)-configuration.Co-expression of St CYP80B with the CYP450 reductases(CPRs,StCPR1,and StCPR2),also cloned from S.tetrandra,significantly enhanced the catalytic activity towards(S)-coclaurine.Site-directed mutagenesis of St CYP80B revealed that the residue H205 is crucial for coclaurine catalysis.Additionally,St CYP80B exhibited tissue-specific expression in plants.This study provides new genetic resources for the biosynthesis of BIAs and further elucidates their synthetic pathway in natural plant systems.
基金supported by the National Natural Science Foundation of China(32130013,32070434)the National Key Research and Development Program of China(2022YFC2601601)+1 种基金the Second Tibetan Plateau Scientific Expedition and Research(STEP)program(2019QZKK05010112,2019QZKK0304-02)Hainan Tropical Rainforest Conservation Research Project,ZDYF2023RDYL01(supported by the Hainan Institute of National Park,HINP,KY-24ZK02).
文摘Understanding the genetic diversity–area relationship(GAR)is essential for comprehending how species adapt to environmental changes,as genetic diversity is an indicator of a species’adaptive potential.Variation in environmental adaptation capacity exists among species and animal taxa with different distribution areas,highlighting the importance of understanding the GAR.To obtain a more comprehensive understanding of the GAR in terrestrial vertebrates,we assessed both haplotype diversity–area and nucleotide diversity–area relationships using 25,453 cytochrome c oxidase subunit I(COI)sequences from 142 amphibian species,574 bird species,and 342 mammal species.We found that both measures of genetic diversity increased with species range size across major animal groups.Nevertheless,the GAR did not differ among animal groups,while haplotype diversity performed better than nucleotide diversity in profiling the GAR,as indicated by higher R2 values.The difference in the modeling fit may stem from the distinct biological and mathematical significance of nucleotide diversity and haplotype diversity.These results suggest that the GAR follows similar rules among different animal taxa.Furthermore,haplotype diversity may serve as a more reliable indicator for assessing the potential effects of area size changes on animal populations and provide better guidance for conserving genetic diversity.
基金supported by the National Key R&D Program of China(No.2021YFC1808902)the National Natural Science Foundation of China(No.31770152)+1 种基金Shaanxi Fundamental Science Research Project for Chemistry&Biology(No.22JHZ008)the Natural Science Foundation of Gansu Province of China(No.23JRRM738).
文摘High molecular weight polycyclic aromatic hydrocarbons(HMW-PAHs)pose significant environmental challenges due to their complex structures and persistent toxicity,and are difficult to be degraded by bacteria.Fungi,however,possess the ability to overcome these challenges,primarily through various enzymes with broad substrate specificity,including cytochrome P450(CYP450),laccase,manganese peroxidase,lignin peroxidase.As a result,the development of efficient fungal strains capable of degrading HMW-PAHs is essential for advancing bioremediation strategies.In this study,Aspergillus fumigatus Z5 was isolated from a contaminated site and demonstrated a remarkable ability to degrade the highly recalcitrant benzo[a]pyrene,achieving a degradation rate of 75.43%.Transcriptomic analysis revealed significant upregulation of 34 CYP450 genes.Among these genes,CYP3A4(gene 8840)showed strong binding affinity for benzo[a]pyrene,as confirmed by molecular docking studies,indicating its key role in the biodegradable process.Furthermore,the metabolic network analysis showed that the electron transfer required for CYP450-mediated oxidative reactions enhanced mitochondrial oxidative phosphorylation and subsequently energy metabolism in A.fumigatus Z5.This metabolic coordination likely facilitates the complete degradation of benzo[a]pyrene.Our study illuminates the fundamental roles of CYP450 from A.fumigatus Z5 in benzo[a]pyrene degradation and provides novel insight into designing and implementing enhanced bioremediation strategies.
基金Natural Science Foundation of Liaoning Province(2022-MS-083)Application Basic Research Plan of Liaoning Province(2023JH2/101300084).
文摘Background:As a major histopathological subtype of gastric cancer(GC),stomach adenocarcinoma(STAD)is an important malignant tumor in the digestive system.Increasing evidence also indicates that endoplasmic reticulum(ER)stress plays a pivotal role in the pathogenesis and progression of GC.Therefore,this study aims to screen and identify vital ER stress-related genes that could contribute to the malignant development and poor prognosis for STAD.Methods:A novel ER stress-related risk score signature was developed employingmachine learning techniques.Then,a prognostic prediction nomogram was also built based on the clinicopathological characteristics and the risk score signature.The tumor immune microenvironment characteristics and pathway enrichment analysis in different risk groups were also explored.Furthermore,through the single-cell RNA sequencing(scRNA-seq)analysis,the study highlightedCytochrome P450 Family 19 SubfamilyAMember 1(CYP19A1)as the pivotal research target and detected its effect on cell proliferation by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide(MTT)and the expression of ER stress-related genes by RT-qPCR in STAD.Results:Based on the evaluation of five screened key ER stressrelated genes(AKR1B1,SERPINE1,ADCYAP1,MATN3,CYP19A1),our ER stress-related risk score signature offers a novel approach for assessing STAD prognosis hazards.The novel prognostic prediction nomogram based on the signature also accurately predicted the survival outcomes of patients with STAD.Furthermore,the expression of CYP19A1 is significantly higher in STAD tissues than in normal tissues.High expression of CYP19A1 was related to a poor survival outcome for patients with STAD.Besides,compared to normal gastric epithelial cells,the expression of CYP19A1 was significantly higher in STAD cell lines.Silencing the expression of CYP19A1 significantly inhibited the cell proliferation ability and decreased the expression of ER stress-related genes,including ATF4,DDIT3 and XBP1 in STAD.Conclusions:In conclusion,our study developed a novel prognosis prediction signature and identified the novel diagnostic and therapeutic target CYP19A1 for patients with STAD.
基金National Natural Science Foundation of China Youth Training Project(2021GZR003)Medical-Engineering Interdisciplinary Research Youth Training Project(2022YGJC001).
文摘Background:Hepatocellular carcinoma(HCC)is a prevalent liver malignancy.This study examined the roles of transforming growth factor beta(TGF-β)and cytochrome b5 domain containing 2(CYB5D2)in HCC etiology and their prognostic biomarker potential.Methods:Key modules and prognostic genes were identified by analyzing the GSE101685 dataset by weighted gene co-expression network analysis(WGCNA)and Least absolute shrinkage and selection operator(LASSO)Cox regression.The expression levels of CYB5D2 and TGF-βin HCC cell lines were quantified using Quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western blotting(WB)assays.Effects of CYB5D2 overexpression on cell proliferation,migration,invasion,and epithelial-mesenchymal transition(EMT)marker regulation were assessed in vitro,while in vivo tumorigenicity was evaluated using a xenograft model of HCC in nude mice.Results:In this study,WGCNA identified the turquoise module as significantly associated with HCC,containing 452 DEGs.LASSO Cox regression analysis revealed 9 key prognostic genes,with CYB5D2 being underexpressed in HCC cells and tissues.TGF-βwas negatively correlated with CYB5D2 expression,resulting in poor patient prognosis.Functional assays demonstrated that CYB5D2 overexpression inhibited proliferation,migration,and invasion of HCC cell lines,and altered EMT marker expression.Furthermore,the addition of TGF-βpartially reversed the suppressive effects caused by CYB5D2 overexpression.In vivo,CYB5D2 overexpression significantly reduced tumor growth,indicating its potential as a therapeutic target for HCC.Conclusion:The tumor suppressor function of CYB5D2 in HCC and its interaction with TGF-βoffered fresh information on the molecular pathophysiology of HCC and possible treatment avenues.
文摘BACKGROUND Kidney transplantation is an effective renal replacement therapy for improving survival and quality of life in chronic kidney disease patients.Kidney transplant recipients need lifelong immunosuppression to prevent rejection and allograft dysfunction.Tacrolimus,a calcineurin inhibitor,is metabolized differently based on cytochrome P4503A(CYP3A)5 genetic variations and this impacts the graft outcome.AIM To examine the clinical outcomes in kidney transplant recipients affected by the variable metabolism of tacrolimus due to the CYP3A5 genetic variation,emphasizing personalized immunosuppression strategies to optimize efficacy,minimize toxicity,and enhance long-term graft survival.METHODS A retrospective study was conducted at a tertiary care center in Central India on 95 kidney transplant recipients.Patient demographics,medical history,CYP3A5 polymorphism,post-transplant investigations,graft biopsy results,preexisting comorbidities,history of post–kidney transplant infections,and new onset diabetes after transplantation(NODAT)was collected.Tacrolimus was initiated at 0.1 mg/kg/day for CYP3A5 expressors and 0.05 mg/kg/day for non-expressors,with dose adjustments to maintain target C0 levels of 7-10 ng/mL for first 6 months and 5-7 ng/mL from 6 months to 12 months posttransplant.Patients were followed regularly for one year for glomerular filtration rate(GFR),creatinine,and the tacrolimus trough concentration(ng/mL)/daily tacrolimus dose(mg/kg/day)ratio(C/D).A P value≤0.05 was considered statistically significant.RESULTS Kidney transplant recipients were classified as expressors(CYP3A51 carriers,n=35)and non-expressors(CYP3A5*3*3,n=60).Both groups were comparable for age,sex,and donor characteristics.Tacrolimus dose was comparable post-transplant except at 6 months and 12 months,where expressors required higher doses.Kidney function(creatinine and estimated GFR),NODAT,hypomagnesemia,and infections showed no significant differences between the two groups over 12 months of follow-up.Biopsy-proven acute rejection(BPAR)was found to be more in expressors(22.9%vs 13.3%,P=0.2340)though it was not found to be statistically significant.Nonexpressors had a significantly higher tacrolimus levels and C/D ratio at multiple follow-ups.CONCLUSION CYP3A5 expressors require higher tacrolimus doses to maintain therapeutic levels as compared to non-expressors.BPAR was higher in expressors but the difference was not significant.Graft function,infection rate,and NODAT were comparable irrespective of CYP3A5 expression status,emphasizing the importance of pretransplant CYP3A5 genotyping and therapeutic drug monitoring to guide tacrolimus dosing for individualized immunosuppressive management.
基金supported by the National Natural Science Foundation of China(Grant Nos.81872831 and 82030107)the National Science and Technology Major Projects for‘significant new drugs creation’of the 13th five-year plan(2017ZX09101001 and 2018ZX09721002007,China)。
文摘D-a-Tocopheryl polyethylene glycol 1000 succinate(TPGS,also known as vitamin E-TPGS)is a biodegradable amphiphilic polymer prepared by esterification of vitamin E with polyethylene glycol(PEG)1000.It is approved by the US Food and Drug Administration(FDA)and has found wide application in nanocarrier drug delivery systems(NDDS).Fully characterizing the in vivo fate and pharmacokinetic behavior of TPGS is important to promote the further development of TPGS-based NDDS.However,to date,a bioassay for the simultaneous quantitation of TPGS and its metabolite,PEG1000,has not been reported.In the present study,we developed such an innovative bioassay and used it to investigate the pharmacokinetics,tissue distribution and excretion of TPGS and PEG1000 in rat after oral and intravenous dosing.In addition,we evaluated the interaction of TPGS with cytochromes P450(CYP450s)in human liver microsomes.The results show that TPGS is poorly absorbed after oral administration with very low bioavailability and that,after intravenous administration,TPGS and PEG1000 are mainly distributed to the spleen,liver,lung and kidney before both being slowly eliminated in urine and feces as PEG1000.In vitro studies show the inhibition of human CYP450 enzymes by TPGS is limited to a weak inhibition of CYP3A4.Overall,our results provide a clear picture of the in vivo fate of TPGS which will be useful in evaluating the safety of TPGS-based NDDS in clinical use and in promoting their further development.
基金This study was supported by the Army Research Office and was accomplished under grant number W911NF‐17‐1‐0345.
文摘Direct interspecies electron transfer(DIET)may be most important in methanogenic environments,but mechanistic studies of DIET to date have primarily focused on cocultures in which fumarate was the terminal electron acceptor.To better understand DIET with methanogens,the transcriptome of Geobacter metallireducens during DIET‐based growth with G.sulfurreducens reducing fumarate was compared with G.metallireducens grown in coculture with diverse Methanosarcina.The transcriptome of G.metallireducens cocultured with G.sulfurreducens was significantly different from those with Methanosarcina.Furthermore,the transcriptome of G.metallireducens grown with Methanosarcina barkeri,which lacks outer‐surface c‐type cytochromes,differed from those of G.metallireducens cocultured with M.acetivorans or M.subterranea,which have an outer‐surface c‐type cytochrome that serves as an electrical connect for DIET.Differences in G.metallireducens expression patterns for genes involved in extracellular electron transfer were particularly notable.Cocultures with c‐type cytochrome deletion mutant strains,ΔGmet_0930,ΔGmet_0557 andΔGmet_2896,never became established with G.sulfurreducens but adapted to grow with all three Methanosarcina.Two porin–cytochrome complexes,PccF and PccG,were important for DIET;however,PccG was more important for growth with Methanosarcina.Unlike cocultures with G.sulfurreducens and M.acetivorans,electrically conductive pili were not needed for growth with M.barkeri.Shewanella oneidensis,another electroactive microbe with abundant outer‐surface c‐type cytochromes,did not grow via DIET.The results demonstrate that the presence of outer‐surface c‐type cytochromes does not necessarily confer the capacity for DIET and emphasize the impact of the electron‐accepting partner on the physiology of the electron‐donating DIET partner.
基金supported by the National Natu-ral Science Foundation of China(Nos.U2006219 and 52101078)China Baowu Low Carbon Metallurgy Innovation Foundation(No.BWLCF202120)+2 种基金the National Key Research and Development Pro-gram of China(No.2020YFA0907300)the Fundamental Research Funds for the Central Universities of the Ministry of Education of China(Nos.N2102009 and N2002019)the Liaoning Revitaliza-tion Talents Program(No.XLYC1907158).
文摘Extracellular electron transfer(EET)plays a critical role in bioelectrochemical processes,allowing cou-pling between microorganisms and extracellular solid-state electrodes,metals,or other cells in energy metabolism.Previous studies have suggested a role for outer-surface c-type cytochromes in direct metal-to-microbe electron transfer by Geobacter sulfurreducens,a model electroactive bacterium.Here,we ex-amined the possibility of other microbially produced electrical contacts by deleting the gene for PilA,the protein monomer that G.sulfurreducens assembles into electrically conductive protein nanowires(e-pili).Deleting pilA gene inhibited electron extraction from pure iron and 316L stainless steel up to 31%and 81%,respectively more than deleting the gene for the outer-surface cytochrome OmcS.This PilA-deficient phenotype,and the observation that relatively thick biofilms(21.7μm)grew on the metal surfaces at multi-cell distances from the metal surfaces suggest that e-pili contributed significantly to microbial cor-rosion via direct metal-to-microbe electron transfer.These results have implications for the fundamental understanding of electron harvest via e-pili by electroactive microbes,their uses in bioenergy production,as well as in monitoring and mitigation of metal biocorrosion.
