The in vitro and in vivo effects of corn peptides(CPs) prepared from corn gluten meal by proteolysis with an alkaline protease and fractions of CPs from Sephadex G-15 and G-10 columns on activities of alcohol dehydr...The in vitro and in vivo effects of corn peptides(CPs) prepared from corn gluten meal by proteolysis with an alkaline protease and fractions of CPs from Sephadex G-15 and G-10 columns on activities of alcohol dehydroge-nase(ADH) were studied. The results show that CPs and fraction 3 of CPs from Sephadex G-10 column enhance in vitro ADH activity. Furthermore, the in vitro accelerating effect of the fraction 3 of CPs on ADH activity was superior to that of glutathione, which was also found even in the presence of ADH inhibitor, such as pyrazole. In the in vivo experiments, the animals were fed with different dosages of CPs and with a dose of Chinese distilled spirit orally, and sacrificed for the measurement of ADH activity. In vivo experimental results indicate that CPS enhanced hepatic ADH activities. To test the safety of CPs as health food, 30 d feeding test was performed. No obvious toxic effects were detected in treated Wistar rats.展开更多
Alcohol dehydrogenase (ADH) catalyzes the interconversion of aldehydes and their corresponding alcohols, and is a key enzyme in volatile ester biosynthesis. However, little is known regarding ADH and ADH encoding ge...Alcohol dehydrogenase (ADH) catalyzes the interconversion of aldehydes and their corresponding alcohols, and is a key enzyme in volatile ester biosynthesis. However, little is known regarding ADH and ADH encoding genes (ADHs) in pear. We identified 8 ADHs in the pear's genome (PbrADHs) by multiple sequences alignment. The PbrADHs were highly ho- mologous in their coding regions, while were diversiform in structure. 9 introns were predicted in PbrADH3-PbrADH8, while 8 introns, generated through exon fusion and intron loss, were predicted in PbrADH1 and PbrADH2. To study the genetic regulation underlying aroma biogenesis in pear fruit, we determined the PbrADH transcripts, ADH activities and volatile contents of fruits during ripening stage for Nanguoli and Dangshansuli, two cultivars having different aroma characteristics. ADH activity was strongly associated with the transcription of ADH~ in the two cultivars during fruit ripening stage. The higher ester content paralleling to a higher ADH activity was detected in Nanguoli than in Dangshansuli, so it is induced that the lower ester content in Dangshansuli fruit may be the result of weak ADH activity. The present study revealed that total ADH activity and volatile ester production correlated with increased PbrADHstranscript levels. PbrADH6 may contribute to ADH activity catalyzing aldehyde reduction and ester formation in pear fruit.展开更多
The activity of alcohol dehydrogenase (ADH) in cultured cells of various tobacco was determined. It was found that significant differences existed in cells of different varieties cultured under normal conditions and a...The activity of alcohol dehydrogenase (ADH) in cultured cells of various tobacco was determined. It was found that significant differences existed in cells of different varieties cultured under normal conditions and as well after treated with exogenous ethanol. The ADH activity had positive relation with the ability of the cells to catabolize exogenous ethanol, indicating that the main function of the ADH in tobacco cells was in the direction of converting ethanol to acetaldehyde.展开更多
Background The most critical mechanism governing drug resistance in Candida albicans (C. albicans) involves efflux pumps, the functionality of which largely depends on energy metabolism. Alcohol dehydrogenase I (A...Background The most critical mechanism governing drug resistance in Candida albicans (C. albicans) involves efflux pumps, the functionality of which largely depends on energy metabolism. Alcohol dehydrogenase I (ADH1) plays an important role in intracellular energy metabolism. The aim of this study was to explore the relationship between ADH1 and drug resistance in C. albicans. Methods Twenty clinical C. albicans samples isolated from individual patients diagnosed with vulvovaginal candidiasis, and two C. albicans strains obtained from a single parental source (the fluconazole (FLC)-sensitive strain CA-1s and the FLC-resistant strain CA-16R) were included in our study. In accordance with the Clinical and Laboratory Standards Institute (CLSI) M27-A3 guidelines, we used the microdilution method to examine the FLC minimum inhibitory concentrations (MICs) and real-time reverse transcription polymerase chain reaction (RT-PCR) to measure the mRNA expression levels of ADH1 and the azole resistance genes CDR1, CDR2, MDR1, FLU1 and ERG11 in all the isolates. Results A highly significant positive correlation between the mRNA levels of ADH1 and the MICs (r =0.921, P=0.000), as well as positive correlations between the mRNA level of ADH1 and those of CDR1, CDR2 and FLU1 (rs of 0.704, 0.772 and 0.779, respectively, P 〈0.01), were observed in the 20 clinical C. albicans samples. The relative expression of ADH1 was upregulated 10.63- to 17.61-fold in all of the drug-resistant isolates. No correlations were found between the mRNA levels of ADH1 and those of MDR1 or ERG11 (P 〉0.05). The mRNA levels of the examined drug resistance genes were higher in the CA-16R strain than in CA-1s, and the mRNA levels of ADH1 in CA-16R were 11.64-fold higher than those in CA-1s (P 〈0.05). Conclusions These results suggest that high levels of ADH1 transcription are implicated in FLC resistance in C. albicans and that the mRNA expression levels of ADH1 are positively correlated with those of CDR1, CDR2 and FLU1.展开更多
Alcohol dehydrogenase 5(ADH5)is a member of medium-chain dehydro-genase/reductase family and takes part in cellular formaldehyde and S-nitrosoglutathione metabolic network.2-tridecanone(2-TD)is a toxic compound in man...Alcohol dehydrogenase 5(ADH5)is a member of medium-chain dehydro-genase/reductase family and takes part in cellular formaldehyde and S-nitrosoglutathione metabolic network.2-tridecanone(2-TD)is a toxic compound in many Solanaceae crops to defend against a variety of herbivory insects.In the broader context of insect development and pest control strategies,this study investigates how a new ADHS from Helicoverpa armigera(HaADH5)regulates the expression of CYP6B6,a gene involved in molting and metamorphosis,in response to 2-TD treatment.Cloning of the HaADH5 complementary DNA sequence revealed that its 1002 bp open reading frame encodes 334 amino acids with a predicted molecular weight of 36.5 kD.HaADH5 protein was purified in the Es-cherichia coli Transetta(pET32a-HaADH5)strain using a prokaryotic expression system.The ability of HaADH5 protein to interact with the 2-TD responsive region within the promoter of CYP6B6 was confirmed by an in vitro electrophoretic mobility shift assay and transcription activity validation in yeast.Finally,the expression levels of both HaADH5 and CYP6B6 were found to be significantly decreased in the midgut of 6th instar larvae after 48 h of treatment with 10 mg/g 2-TD artificial diet.These results indicate that upon 2-TD treatment of cotton bollworm,HaADHS regulates the expression of CYP6B6 by interacting with its promoter.As HaADH5 regulation of CYP6B6 expression may con-tribute to the larval xenobiotic detoxification,molting and metamorphosis,HaADH5 is a.candidate target for controlling the growth and development of cotton bollworm.展开更多
The catalytic characteristics of horse liver alcohol dehydrogenase (HLADH) in the systems involving ionic liquids (ILs) (BMIm·Cl, BMIm·Br, BMIm·PF6, BMIm·BF4 BMIm·OTf and EMIm·Cl) w...The catalytic characteristics of horse liver alcohol dehydrogenase (HLADH) in the systems involving ionic liquids (ILs) (BMIm·Cl, BMIm·Br, BMIm·PF6, BMIm·BF4 BMIm·OTf and EMIm·Cl) were examined. HLADH displayed higher oxidation activity towards ethanol in the systems containing BMIm·Cl, BMIm·Br, EMIm·Cl or BMIm·PF6 with proper content than that in the IL-free buffer. An excessive amount of these ILs in the reaction systems resulted in an obvious decline in enzymatic activity. BMIm·BF4 and BMIm·OTf of any content investigated could considerably inhibit the enzyme. The anions of ILs showed significant effect on the activity, kinetic parameters and activation energy of HLADH-mediated ethanol oxidation. Additionally, BMIm·Cl, BMIm·Br, EMIm·Cl and BMIm·PF6 boosted markedly the thermostability of HLADH, while the enzyme was less thermostable in BMIm·BF4 or BMIm·OTf-containing systems. The associated conformational changes in HLADH caused by ILs were examined by UV technique.展开更多
Background Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase 2 (ALDH2) are the key en- zymes for alcohol metabolism. Several genetic studies have investigated the association between ADH and ALDH2 genetic pol...Background Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase 2 (ALDH2) are the key en- zymes for alcohol metabolism. Several genetic studies have investigated the association between ADH and ALDH2 genetic polymorphisms and serum lipid profile (SLP), however, the results were inconsistent. Methods Fourteen articles involving 27,917 participants were included in this meta-analysis. Weighted mean difference (WMD) and 95% confidence intervals (CIs) were presented using random effects model. Publication bias was evaluated by funnel plot and Begg's test. In addition, to further explore the heterogeneity, subgroup analysis were performed. Results Overall, there was no association between ADH genetic polymorphisms and SLP with no regard for drinking status. However, compared with ALDH2 wild homozygous genotype, ALDH2 mutant genotypes were associated with significant decrease in serum high density lipoprotein cholesterol (HDL- C) (WMD -1.80 mg/dL, 95% CI -1.88 to -1.72, P 〈 0.001) and total cholesterol (TC) levels (WMD -1.]0 mg/dL, 95% CI -1.59 to -0.62, P 〈 0.001), and significant increase in serum low density lipoprotein choles- terol (LDL-C) level (WMD 0.30 mg/dL, 95% CI 0.18 to 0.43, P 〈 0.001). Although there was no significant difference in serum triglyceride level in the overall population, subgroup analysis revealed that compared with ALDH2 * 1 wild homozygote, ALDH2 * 2 allele displayed a significant difference in serum triglyceride level between the female and male (female: WMD 1.69 mg/dl, 95% CI 1.08 to 2.30, P 〈 0.001; male: WMD -6.42 mg/dL, 95% CI -12.15 to -0.68, P = 0.028). Conclusion ADH genetic polymorphism has no association with SLP, regardless of sex category and drinking status. ALDH2 genetic polymorphism has slight association with HDL-C, LDL-C and TC levels and sex-specific association with serum triglyceride level. Whether or not the association between ADH2 genetic polymorphisms and SLP is resulted from alcohol con-sumption needs further investigation.展开更多
AIM To compare transcriptomes of non-alcoholic fatty liver disease(NAFLD) and alcoholic liver disease(ALD) in a meta-analysis of liver biopsies.METHODS Employing transcriptome data from patient liver biopsies retrieve...AIM To compare transcriptomes of non-alcoholic fatty liver disease(NAFLD) and alcoholic liver disease(ALD) in a meta-analysis of liver biopsies.METHODS Employing transcriptome data from patient liver biopsies retrieved from several public repositories we performed a meta-analysis comparing ALD and NAFLD.RESULTS We observed predominating commonalities at the transcriptome level between ALD and NAFLD,most prominently numerous down-regulated metabolic pathways and cytochrome-related pathways and a few up-regulated pathways which include ECM-receptor interaction,phagosome and lysosome.However some pathways were regulated in opposite directions in ALD and NAFLD,for example,glycolysis was down-regulated in ALD and up-regulated in NAFLD.Interestingly,we found rate-limiting genes such as HMGCR,SQLE and CYP7A1 which are associated with cholesterol processes adversely regulated between ALD(down-regulated) and NAFLD(up-regulated).We propose that similar phenotypes in both diseases may be due to a lower level of the enzyme CYP7A1 compared to the cholesterol synthesis enzymes HMGCR and SQLE.Additionally,we provide a compendium of comparative KEGG pathways regulation in ALD and NAFLD.CONCLUSION Our finding of adversely regulated cholesterol processes in ALD and NAFLD draws the focus to regulation of cholesterol secretion into bile.Thus,it will be interesting to further investigate CYP7A1-mediated cholesterol secretion into bile-also as possible drug targets.The list of potential novel biomarkers may assist differential diagnosis of ALD and NAFLD.展开更多
It is well known that exposure to environmental stresses could enhance the adaptability of bacteria and up-regulate the expression of a variety of oxidative stress-related genes and antioxidant enzymes.It is unclear w...It is well known that exposure to environmental stresses could enhance the adaptability of bacteria and up-regulate the expression of a variety of oxidative stress-related genes and antioxidant enzymes.It is unclear whether the adaptability of microorganisms formed naturally in special environments could transfer to other organisms.The study aimed to evaluate the effects of untreated and ethanol-induced Lactococcus lactis intracellular extracts(U-IE and E-IE)on alcohol metabolism in mice.The positive effects of E-IE on alcohol metabolism in mice were revealed by the enhanced latency of loss of righting reflex(LORR),the reduced duration of LORR,the decrease of blood alcohol concentration,as well as the elevation of alcohol dehydrogenase(ADH)activities in the stomach and liver tissues.Furthermore,the potential benefits of E-IE on the liver were evaluated by biochemical parameters including the activities of serum transaminase,the levels of antioxidant enzymes,and the pathological changes of liver tissue.The present work put forward a new point that appropriate ethanol stress could enhance the intracellular ADH activity of L.lactis,and its intracellular extracts could continue to enhance alcohol metabolism in mice.展开更多
Hyperthermus butylicus is a hyperthermophilic crenarchaeon that produces 1-butanol as an end product.A thermostable alcohol dehydrogenase(ADH)must be present in H.butylicus to act as the key enzyme responsible for thi...Hyperthermus butylicus is a hyperthermophilic crenarchaeon that produces 1-butanol as an end product.A thermostable alcohol dehydrogenase(ADH)must be present in H.butylicus to act as the key enzyme responsible for this production;however,the gene that encodes the ADH has not yet been identified.A novel ADH,HbADH2,was purified from a cell-free extract of H.butylicus,and its characteristics were determined.The gene that encodes HbADH2 was demonstrated to be HBUT_RS04850 and annotated as a hypothetical protein in H.butylicus.HbADH2 was found to be a primary-secondary ADH capable of using a wide range of substrates,including butyraldehyde and butanol.Butyraldehyde had the highest specificity constant,calculated as k_(cat)/K_(m),with kcat and apparent K_(m) values of 8.00±0.22 s^(-1) and 0.59±0.07 mM,respectively.The apparent Km values for other substrates,including ethanol,1-propanol,2-propanol,butanol,acetaldehyde,propanal,and acetone,were 4.36±0.42,4.69±0.41,3.74±0.46,2.44±0.30,1.27±0.18,1.55±0.20,and 0.68±0.04 mM,respectively.The optimal pH values for catalyzing aldehyde reduction and alcohol oxidation were 6.0 and 9.0,respectively,while the optimal temperature was higher than 90°C due to the increase in enzymatic activity from 60℃ to 90℃.Based on its substrate specificity,enzyme kinetics,and thermostability,HbADH2 may be the ADH that catalyzes the production of 1-butanol in H.butylicus.The putative conserved motif sites for NAD(P)^(+)and iron binding were identified by aligning HbADH2 with previously characterized Fe-containing ADHs.展开更多
Our previous studies have revealed that amyloidβ(Aβ)-binding alcohol dehydrogenase (ABAD) decoy peptide antagonizes Aβ42-induced neurotoxicity. However, whether it improves oxidative stress injury remains uncle...Our previous studies have revealed that amyloidβ(Aβ)-binding alcohol dehydrogenase (ABAD) decoy peptide antagonizes Aβ42-induced neurotoxicity. However, whether it improves oxidative stress injury remains unclear. In this study, a recombinant adenovirus constitutively secreting and expressing Aβ-ABAD decoy peptide (rAAV/ABAD-DP-6His) was successfully constructed. Our results showed that rAAV/ABAD-DP-6His increased superoxide dismutase activity in hydro-gen peroxide-induced oxidative stress-mediated injury of PC12 cells. Moreover, rAAV/ABAD-DP-6His decreased malondialdehyde content, intracellular Ca2+concentration, and the level of reactive oxygen species. rAAV/ABAD-DP-6His maintained the stability of the mitochondrial membrane potential. In addition, the ATP level remained constant, and apoptosis was reduced. Overall, the results indicate that rAAV/ABAD-DP-6His generates the fusion peptide, Aβ-ABAD decoy peptide, which effectively protects PC12 cells from oxidative stress injury induced by hy-drogen peroxide, thus exerting neuroprotective effects.展开更多
Amyloid β peptide binding alcohol dehydrogenase (ABAD) decoy peptide (DP) can competitively antagonize binding of amyloid β peptide to ABAD and inhibit the cytotoxic effects of amyloid β peptide. Based on pepti...Amyloid β peptide binding alcohol dehydrogenase (ABAD) decoy peptide (DP) can competitively antagonize binding of amyloid β peptide to ABAD and inhibit the cytotoxic effects of amyloid β peptide. Based on peptide aptamers, the present study inserted ABAD-DP into the disulfide bond of human thioredoxin (TRX) using molecular cloning technique to construct a fusion gene that can express the TRX1-ABAD-DP-TRX2 aptamer. Moreover, adeno-associated virus was used to allow its stable expression. Immunofluorescent staining revealed the co-expression of the transduced fusion gene TRX1-ABAD-DP-TRX2 and amyloid β peptide in NIH-3T3 cells, indicating that the TRXl-ABAD-DP-TRX2 aptamer can bind amyloid β peptide within cells. In addition, cell morphology and MTT results suggested that TRX1-ABAD-DP-TRX2 attenuated amyloid β peptide-induced SH-SY5Y cell injury and improved cell viability. These findings confirmed the possibility of constructing TRX-based peptide aptamer using ABAD-DP. Moreover, TRXl-ABAD-DP-TRX2 inhibited the cytotoxic effect of amyloid β peptide.展开更多
Objective:To explore the expression and prognostic significance of ADHs in Hepatocellular carcinoma(HCC).Methods:The clinical data in this study were obtained from The Cancer Genome Atlas(TCGA).The expression of ADHs ...Objective:To explore the expression and prognostic significance of ADHs in Hepatocellular carcinoma(HCC).Methods:The clinical data in this study were obtained from The Cancer Genome Atlas(TCGA).The expression of ADHs was differentially analyzed in normal liver tissues and HCC using the Metabolic gEne RApid Visualizer and the TCGA database,and the differentially expressed ADHs were selected for kaplan-meier survival analysis.Cox analysis was performed to select factors that may influence the prognosis of HCC and to verify independent risk factors for HCC patients.The interaction among ADHs was explored at the gene level and protein expression level through GeneMAMIA and STRING,and the functional enrichment analysis of ADH family was carried out using DAVID bioinformatics resources.Results:The expression of ADH1A,ADH1B,ADH1C,ADH4 and ADH7 in HCC is low.The low-expression groups of ADH1A,ADH1B,ADH1C and ADH4 had poor survival rates,which may be related to the poor prognosis of HCC patients.Univariate Cox regression analysis showed that the expression levels of ADH1A,ADH1C and ADH4,as well as the clinical stage,T stage and M stage of the tumor were closely related to the overall survival rate of the patients.Multivariate Cox regression analysis further suggested that the low expression of ADH1A,ADH1C and ADH4 was an independent risk factor affecting the prognosis of HCC patients.There is a pathway between ADH1A-ADH1B,ADH1B-ADH1C,ADH1A-ADH1C,and the ADHs is closely related to Esterase D and Aldehyde dehydrogenase.The ADHs is mainly involved in biological processes such as Ethanol oxidation and Retinol metabolism,and plays a biological role in Glycolysis/Gluconeogenesis,Chemical carcinogenesis and Metabolism of xenobiotics by cytochrome P450.Conclusion:ADH1A,ADH1C and ADH4 may be biomarkers for the prognosis of HCC,providing reference value for the practical application of ADHs in HCC.展开更多
Cinnamoyl CoA reductase (CCR) and cinnamyl alcohol dehydrogenase (CAD) catalyze the last steps of mono- lignol biosynthesis. In Arabidopsis, one CCR gene (CCR1, Atlg15950) and two CAD genes (CAD C At3g19450 and...Cinnamoyl CoA reductase (CCR) and cinnamyl alcohol dehydrogenase (CAD) catalyze the last steps of mono- lignol biosynthesis. In Arabidopsis, one CCR gene (CCR1, Atlg15950) and two CAD genes (CAD C At3g19450 and CAD D At4g34230) are involved in this pathway. A triple cad c cad d ccrl mutant, named ccc, was obtained. This mutant displays a severe dwarf phenotype and male sterility. The lignin content in ccc mature stems is reduced to 50% of the wild-type level. In addition, stem lignin structure is severely affected, as shown by the dramatic enrichment in resistant inter-unit bonds and incorporation into the polymer of monolignol precursors such as coniferaldehyde, sinapaldehyde, and ferulic acid. Male sterility is due to the lack of lignification in the anther endothecium, which causes the failure of anther de- hiscence and of pollen release. The cc~ hypolignified stems accumulate higher amounts of flavonol glycosides, sinapoyl malate and feruloyl malate, which suggests a redirection of the phenolic pathway. Therefore, the absence of CAD and CCR, key enzymes of the monolignol pathway, has more severe consequences on the phenotype than the individual absence of each of them. Induction of another CCR (CCR2, Atlg80820) and another CAD (CAD1, At4g39330) does not compensate the absence of the main CCR and CAD activities. This lack of CCR and CAD activities not only impacts lignification, but also severely affects the development of the plants. These consequences must be carefully considered when trying to reduce the lignin content of plants in order to facilitate the lignocellulose-to-bioethanol conversion process.展开更多
The presence of lignin reduces the quality of lignocellulosic biomass for forage materials and feedstock for biofuels. In C4 grasses, the brown midrib phenotype has been linked to mutations to genes in the monolignol ...The presence of lignin reduces the quality of lignocellulosic biomass for forage materials and feedstock for biofuels. In C4 grasses, the brown midrib phenotype has been linked to mutations to genes in the monolignol biosynthesis pathway. For example, the Bmr6 gene in sorghum (Sorghum bicolor) has been previously shown to encode cinnamyl alcohol dehydrogenase (CAD), which catalyzes the final step of the monolignol biosynthesis pathway. Mutations in this gene have been shown to reduce the abundance of lignin, enhance digestibility, and improve saccharification efficiencies and ethanol yields. Nine sorghum lines harboring five different bmr6 alleles were identified in an EMS-mutagenized TILLING population. DNA sequencing of Bmr6 revealed that the majority of the mutations impacted evolutionarily conserved amino acids while three-dimensional structural modeling predicted that all of these alleles interfered with the enzyme's ability to bind with its NAPPH cofactor. All of the new alleles reduced in vitro CAD activity levels and enhanced glucose yields following saccharification. Further, many of these lines were associated with higher reductions in acid detergent lignin compared to lines harboring the previously characterized bmr6-ref allele. These bmr6 lines represent new breeding tools for manipulating biomass composition to enhance forage and feedstock quality.展开更多
基金Supported by the Science Research Foundation of Jilin Agricultural University, China(No.205-00370)
文摘The in vitro and in vivo effects of corn peptides(CPs) prepared from corn gluten meal by proteolysis with an alkaline protease and fractions of CPs from Sephadex G-15 and G-10 columns on activities of alcohol dehydroge-nase(ADH) were studied. The results show that CPs and fraction 3 of CPs from Sephadex G-10 column enhance in vitro ADH activity. Furthermore, the in vitro accelerating effect of the fraction 3 of CPs on ADH activity was superior to that of glutathione, which was also found even in the presence of ADH inhibitor, such as pyrazole. In the in vivo experiments, the animals were fed with different dosages of CPs and with a dose of Chinese distilled spirit orally, and sacrificed for the measurement of ADH activity. In vivo experimental results indicate that CPS enhanced hepatic ADH activities. To test the safety of CPs as health food, 30 d feeding test was performed. No obvious toxic effects were detected in treated Wistar rats.
基金financially supported by the National Natural Science Foundation of China(31301739)the Key Technologies R&D Program of China during the 12th Five-year Plan Period(2013BAD02B01-4)
文摘Alcohol dehydrogenase (ADH) catalyzes the interconversion of aldehydes and their corresponding alcohols, and is a key enzyme in volatile ester biosynthesis. However, little is known regarding ADH and ADH encoding genes (ADHs) in pear. We identified 8 ADHs in the pear's genome (PbrADHs) by multiple sequences alignment. The PbrADHs were highly ho- mologous in their coding regions, while were diversiform in structure. 9 introns were predicted in PbrADH3-PbrADH8, while 8 introns, generated through exon fusion and intron loss, were predicted in PbrADH1 and PbrADH2. To study the genetic regulation underlying aroma biogenesis in pear fruit, we determined the PbrADH transcripts, ADH activities and volatile contents of fruits during ripening stage for Nanguoli and Dangshansuli, two cultivars having different aroma characteristics. ADH activity was strongly associated with the transcription of ADH~ in the two cultivars during fruit ripening stage. The higher ester content paralleling to a higher ADH activity was detected in Nanguoli than in Dangshansuli, so it is induced that the lower ester content in Dangshansuli fruit may be the result of weak ADH activity. The present study revealed that total ADH activity and volatile ester production correlated with increased PbrADHstranscript levels. PbrADH6 may contribute to ADH activity catalyzing aldehyde reduction and ester formation in pear fruit.
文摘The activity of alcohol dehydrogenase (ADH) in cultured cells of various tobacco was determined. It was found that significant differences existed in cells of different varieties cultured under normal conditions and as well after treated with exogenous ethanol. The ADH activity had positive relation with the ability of the cells to catabolize exogenous ethanol, indicating that the main function of the ADH in tobacco cells was in the direction of converting ethanol to acetaldehyde.
基金This study was supported by grants from the National Natural Science Foundation of China (Nos. 81171542, 30972660), the Natural Science Foundation of Guangdong Province, China (No. 10151008901000131), Guangzhou Key Technology RD Program, China (No. 2010J-E011), and the Fundamental Research Funds for the Central Universities (No. 21611509).
文摘Background The most critical mechanism governing drug resistance in Candida albicans (C. albicans) involves efflux pumps, the functionality of which largely depends on energy metabolism. Alcohol dehydrogenase I (ADH1) plays an important role in intracellular energy metabolism. The aim of this study was to explore the relationship between ADH1 and drug resistance in C. albicans. Methods Twenty clinical C. albicans samples isolated from individual patients diagnosed with vulvovaginal candidiasis, and two C. albicans strains obtained from a single parental source (the fluconazole (FLC)-sensitive strain CA-1s and the FLC-resistant strain CA-16R) were included in our study. In accordance with the Clinical and Laboratory Standards Institute (CLSI) M27-A3 guidelines, we used the microdilution method to examine the FLC minimum inhibitory concentrations (MICs) and real-time reverse transcription polymerase chain reaction (RT-PCR) to measure the mRNA expression levels of ADH1 and the azole resistance genes CDR1, CDR2, MDR1, FLU1 and ERG11 in all the isolates. Results A highly significant positive correlation between the mRNA levels of ADH1 and the MICs (r =0.921, P=0.000), as well as positive correlations between the mRNA level of ADH1 and those of CDR1, CDR2 and FLU1 (rs of 0.704, 0.772 and 0.779, respectively, P 〈0.01), were observed in the 20 clinical C. albicans samples. The relative expression of ADH1 was upregulated 10.63- to 17.61-fold in all of the drug-resistant isolates. No correlations were found between the mRNA levels of ADH1 and those of MDR1 or ERG11 (P 〉0.05). The mRNA levels of the examined drug resistance genes were higher in the CA-16R strain than in CA-1s, and the mRNA levels of ADH1 in CA-16R were 11.64-fold higher than those in CA-1s (P 〈0.05). Conclusions These results suggest that high levels of ADH1 transcription are implicated in FLC resistance in C. albicans and that the mRNA expression levels of ADH1 are positively correlated with those of CDR1, CDR2 and FLU1.
基金This work was supported by the National Natural Science Foundation of China(31471781)Natural Science Foun-dation of Xinjiang Region in China(2016D01C042)Tianshan Cedar Project in 2017(2017xs20).
文摘Alcohol dehydrogenase 5(ADH5)is a member of medium-chain dehydro-genase/reductase family and takes part in cellular formaldehyde and S-nitrosoglutathione metabolic network.2-tridecanone(2-TD)is a toxic compound in many Solanaceae crops to defend against a variety of herbivory insects.In the broader context of insect development and pest control strategies,this study investigates how a new ADHS from Helicoverpa armigera(HaADH5)regulates the expression of CYP6B6,a gene involved in molting and metamorphosis,in response to 2-TD treatment.Cloning of the HaADH5 complementary DNA sequence revealed that its 1002 bp open reading frame encodes 334 amino acids with a predicted molecular weight of 36.5 kD.HaADH5 protein was purified in the Es-cherichia coli Transetta(pET32a-HaADH5)strain using a prokaryotic expression system.The ability of HaADH5 protein to interact with the 2-TD responsive region within the promoter of CYP6B6 was confirmed by an in vitro electrophoretic mobility shift assay and transcription activity validation in yeast.Finally,the expression levels of both HaADH5 and CYP6B6 were found to be significantly decreased in the midgut of 6th instar larvae after 48 h of treatment with 10 mg/g 2-TD artificial diet.These results indicate that upon 2-TD treatment of cotton bollworm,HaADHS regulates the expression of CYP6B6 by interacting with its promoter.As HaADH5 regulation of CYP6B6 expression may con-tribute to the larval xenobiotic detoxification,molting and metamorphosis,HaADH5 is a.candidate target for controlling the growth and development of cotton bollworm.
基金Project supported by the National Natural Science Foundation of China (No. 20406006) and Department of Education of Fujian Province (No. JB04028).
文摘The catalytic characteristics of horse liver alcohol dehydrogenase (HLADH) in the systems involving ionic liquids (ILs) (BMIm·Cl, BMIm·Br, BMIm·PF6, BMIm·BF4 BMIm·OTf and EMIm·Cl) were examined. HLADH displayed higher oxidation activity towards ethanol in the systems containing BMIm·Cl, BMIm·Br, EMIm·Cl or BMIm·PF6 with proper content than that in the IL-free buffer. An excessive amount of these ILs in the reaction systems resulted in an obvious decline in enzymatic activity. BMIm·BF4 and BMIm·OTf of any content investigated could considerably inhibit the enzyme. The anions of ILs showed significant effect on the activity, kinetic parameters and activation energy of HLADH-mediated ethanol oxidation. Additionally, BMIm·Cl, BMIm·Br, EMIm·Cl and BMIm·PF6 boosted markedly the thermostability of HLADH, while the enzyme was less thermostable in BMIm·BF4 or BMIm·OTf-containing systems. The associated conformational changes in HLADH caused by ILs were examined by UV technique.
基金supported by National Natural Science Foundation of China(No.81301676)the PhD Start-up Program of Guangdong Province Natural Science Fund(No.S2012040008047)
文摘Background Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase 2 (ALDH2) are the key en- zymes for alcohol metabolism. Several genetic studies have investigated the association between ADH and ALDH2 genetic polymorphisms and serum lipid profile (SLP), however, the results were inconsistent. Methods Fourteen articles involving 27,917 participants were included in this meta-analysis. Weighted mean difference (WMD) and 95% confidence intervals (CIs) were presented using random effects model. Publication bias was evaluated by funnel plot and Begg's test. In addition, to further explore the heterogeneity, subgroup analysis were performed. Results Overall, there was no association between ADH genetic polymorphisms and SLP with no regard for drinking status. However, compared with ALDH2 wild homozygous genotype, ALDH2 mutant genotypes were associated with significant decrease in serum high density lipoprotein cholesterol (HDL- C) (WMD -1.80 mg/dL, 95% CI -1.88 to -1.72, P 〈 0.001) and total cholesterol (TC) levels (WMD -1.]0 mg/dL, 95% CI -1.59 to -0.62, P 〈 0.001), and significant increase in serum low density lipoprotein choles- terol (LDL-C) level (WMD 0.30 mg/dL, 95% CI 0.18 to 0.43, P 〈 0.001). Although there was no significant difference in serum triglyceride level in the overall population, subgroup analysis revealed that compared with ALDH2 * 1 wild homozygote, ALDH2 * 2 allele displayed a significant difference in serum triglyceride level between the female and male (female: WMD 1.69 mg/dl, 95% CI 1.08 to 2.30, P 〈 0.001; male: WMD -6.42 mg/dL, 95% CI -12.15 to -0.68, P = 0.028). Conclusion ADH genetic polymorphism has no association with SLP, regardless of sex category and drinking status. ALDH2 genetic polymorphism has slight association with HDL-C, LDL-C and TC levels and sex-specific association with serum triglyceride level. Whether or not the association between ADH2 genetic polymorphisms and SLP is resulted from alcohol con-sumption needs further investigation.
基金Supported by The Medical Faculty of the Heinrich Heine University Düsseldorf
文摘AIM To compare transcriptomes of non-alcoholic fatty liver disease(NAFLD) and alcoholic liver disease(ALD) in a meta-analysis of liver biopsies.METHODS Employing transcriptome data from patient liver biopsies retrieved from several public repositories we performed a meta-analysis comparing ALD and NAFLD.RESULTS We observed predominating commonalities at the transcriptome level between ALD and NAFLD,most prominently numerous down-regulated metabolic pathways and cytochrome-related pathways and a few up-regulated pathways which include ECM-receptor interaction,phagosome and lysosome.However some pathways were regulated in opposite directions in ALD and NAFLD,for example,glycolysis was down-regulated in ALD and up-regulated in NAFLD.Interestingly,we found rate-limiting genes such as HMGCR,SQLE and CYP7A1 which are associated with cholesterol processes adversely regulated between ALD(down-regulated) and NAFLD(up-regulated).We propose that similar phenotypes in both diseases may be due to a lower level of the enzyme CYP7A1 compared to the cholesterol synthesis enzymes HMGCR and SQLE.Additionally,we provide a compendium of comparative KEGG pathways regulation in ALD and NAFLD.CONCLUSION Our finding of adversely regulated cholesterol processes in ALD and NAFLD draws the focus to regulation of cholesterol secretion into bile.Thus,it will be interesting to further investigate CYP7A1-mediated cholesterol secretion into bile-also as possible drug targets.The list of potential novel biomarkers may assist differential diagnosis of ALD and NAFLD.
基金supported by the National Natural Science Foundation of China(81870093)the Research Project of People’s Liberation Army(BX115C007,BFP20C006)+1 种基金the Special Subject Funding of Zhengzhou Universitythe Natural Science Foundation of Henan Province for Outstanding Youth(202300410365)。
文摘It is well known that exposure to environmental stresses could enhance the adaptability of bacteria and up-regulate the expression of a variety of oxidative stress-related genes and antioxidant enzymes.It is unclear whether the adaptability of microorganisms formed naturally in special environments could transfer to other organisms.The study aimed to evaluate the effects of untreated and ethanol-induced Lactococcus lactis intracellular extracts(U-IE and E-IE)on alcohol metabolism in mice.The positive effects of E-IE on alcohol metabolism in mice were revealed by the enhanced latency of loss of righting reflex(LORR),the reduced duration of LORR,the decrease of blood alcohol concentration,as well as the elevation of alcohol dehydrogenase(ADH)activities in the stomach and liver tissues.Furthermore,the potential benefits of E-IE on the liver were evaluated by biochemical parameters including the activities of serum transaminase,the levels of antioxidant enzymes,and the pathological changes of liver tissue.The present work put forward a new point that appropriate ethanol stress could enhance the intracellular ADH activity of L.lactis,and its intracellular extracts could continue to enhance alcohol metabolism in mice.
基金supported by a research grant from the Natural Sciences and Engineering Research Council of Canada(NSERC)to K.M.
文摘Hyperthermus butylicus is a hyperthermophilic crenarchaeon that produces 1-butanol as an end product.A thermostable alcohol dehydrogenase(ADH)must be present in H.butylicus to act as the key enzyme responsible for this production;however,the gene that encodes the ADH has not yet been identified.A novel ADH,HbADH2,was purified from a cell-free extract of H.butylicus,and its characteristics were determined.The gene that encodes HbADH2 was demonstrated to be HBUT_RS04850 and annotated as a hypothetical protein in H.butylicus.HbADH2 was found to be a primary-secondary ADH capable of using a wide range of substrates,including butyraldehyde and butanol.Butyraldehyde had the highest specificity constant,calculated as k_(cat)/K_(m),with kcat and apparent K_(m) values of 8.00±0.22 s^(-1) and 0.59±0.07 mM,respectively.The apparent Km values for other substrates,including ethanol,1-propanol,2-propanol,butanol,acetaldehyde,propanal,and acetone,were 4.36±0.42,4.69±0.41,3.74±0.46,2.44±0.30,1.27±0.18,1.55±0.20,and 0.68±0.04 mM,respectively.The optimal pH values for catalyzing aldehyde reduction and alcohol oxidation were 6.0 and 9.0,respectively,while the optimal temperature was higher than 90°C due to the increase in enzymatic activity from 60℃ to 90℃.Based on its substrate specificity,enzyme kinetics,and thermostability,HbADH2 may be the ADH that catalyzes the production of 1-butanol in H.butylicus.The putative conserved motif sites for NAD(P)^(+)and iron binding were identified by aligning HbADH2 with previously characterized Fe-containing ADHs.
基金supported by the National Natural Science Foundation of China for the Youth,No.30800338,30801211the National Natural Science Foundation of China,No.30872721
文摘Our previous studies have revealed that amyloidβ(Aβ)-binding alcohol dehydrogenase (ABAD) decoy peptide antagonizes Aβ42-induced neurotoxicity. However, whether it improves oxidative stress injury remains unclear. In this study, a recombinant adenovirus constitutively secreting and expressing Aβ-ABAD decoy peptide (rAAV/ABAD-DP-6His) was successfully constructed. Our results showed that rAAV/ABAD-DP-6His increased superoxide dismutase activity in hydro-gen peroxide-induced oxidative stress-mediated injury of PC12 cells. Moreover, rAAV/ABAD-DP-6His decreased malondialdehyde content, intracellular Ca2+concentration, and the level of reactive oxygen species. rAAV/ABAD-DP-6His maintained the stability of the mitochondrial membrane potential. In addition, the ATP level remained constant, and apoptosis was reduced. Overall, the results indicate that rAAV/ABAD-DP-6His generates the fusion peptide, Aβ-ABAD decoy peptide, which effectively protects PC12 cells from oxidative stress injury induced by hy-drogen peroxide, thus exerting neuroprotective effects.
基金supported by the National Natural Science Foundation of China,No.30872721the National Natural Science Foundation for the Youth,No.30801211,30800338the Scientific Research Foundation for New Teachers of High Institutes,No.200801831073,200801831072
文摘Amyloid β peptide binding alcohol dehydrogenase (ABAD) decoy peptide (DP) can competitively antagonize binding of amyloid β peptide to ABAD and inhibit the cytotoxic effects of amyloid β peptide. Based on peptide aptamers, the present study inserted ABAD-DP into the disulfide bond of human thioredoxin (TRX) using molecular cloning technique to construct a fusion gene that can express the TRX1-ABAD-DP-TRX2 aptamer. Moreover, adeno-associated virus was used to allow its stable expression. Immunofluorescent staining revealed the co-expression of the transduced fusion gene TRX1-ABAD-DP-TRX2 and amyloid β peptide in NIH-3T3 cells, indicating that the TRXl-ABAD-DP-TRX2 aptamer can bind amyloid β peptide within cells. In addition, cell morphology and MTT results suggested that TRX1-ABAD-DP-TRX2 attenuated amyloid β peptide-induced SH-SY5Y cell injury and improved cell viability. These findings confirmed the possibility of constructing TRX-based peptide aptamer using ABAD-DP. Moreover, TRXl-ABAD-DP-TRX2 inhibited the cytotoxic effect of amyloid β peptide.
基金supported by Wuhan Municipal Project of Scientific Problem Tackling(No.201161038342-02)。
文摘Objective:To explore the expression and prognostic significance of ADHs in Hepatocellular carcinoma(HCC).Methods:The clinical data in this study were obtained from The Cancer Genome Atlas(TCGA).The expression of ADHs was differentially analyzed in normal liver tissues and HCC using the Metabolic gEne RApid Visualizer and the TCGA database,and the differentially expressed ADHs were selected for kaplan-meier survival analysis.Cox analysis was performed to select factors that may influence the prognosis of HCC and to verify independent risk factors for HCC patients.The interaction among ADHs was explored at the gene level and protein expression level through GeneMAMIA and STRING,and the functional enrichment analysis of ADH family was carried out using DAVID bioinformatics resources.Results:The expression of ADH1A,ADH1B,ADH1C,ADH4 and ADH7 in HCC is low.The low-expression groups of ADH1A,ADH1B,ADH1C and ADH4 had poor survival rates,which may be related to the poor prognosis of HCC patients.Univariate Cox regression analysis showed that the expression levels of ADH1A,ADH1C and ADH4,as well as the clinical stage,T stage and M stage of the tumor were closely related to the overall survival rate of the patients.Multivariate Cox regression analysis further suggested that the low expression of ADH1A,ADH1C and ADH4 was an independent risk factor affecting the prognosis of HCC patients.There is a pathway between ADH1A-ADH1B,ADH1B-ADH1C,ADH1A-ADH1C,and the ADHs is closely related to Esterase D and Aldehyde dehydrogenase.The ADHs is mainly involved in biological processes such as Ethanol oxidation and Retinol metabolism,and plays a biological role in Glycolysis/Gluconeogenesis,Chemical carcinogenesis and Metabolism of xenobiotics by cytochrome P450.Conclusion:ADH1A,ADH1C and ADH4 may be biomarkers for the prognosis of HCC,providing reference value for the practical application of ADHs in HCC.
文摘Cinnamoyl CoA reductase (CCR) and cinnamyl alcohol dehydrogenase (CAD) catalyze the last steps of mono- lignol biosynthesis. In Arabidopsis, one CCR gene (CCR1, Atlg15950) and two CAD genes (CAD C At3g19450 and CAD D At4g34230) are involved in this pathway. A triple cad c cad d ccrl mutant, named ccc, was obtained. This mutant displays a severe dwarf phenotype and male sterility. The lignin content in ccc mature stems is reduced to 50% of the wild-type level. In addition, stem lignin structure is severely affected, as shown by the dramatic enrichment in resistant inter-unit bonds and incorporation into the polymer of monolignol precursors such as coniferaldehyde, sinapaldehyde, and ferulic acid. Male sterility is due to the lack of lignification in the anther endothecium, which causes the failure of anther de- hiscence and of pollen release. The cc~ hypolignified stems accumulate higher amounts of flavonol glycosides, sinapoyl malate and feruloyl malate, which suggests a redirection of the phenolic pathway. Therefore, the absence of CAD and CCR, key enzymes of the monolignol pathway, has more severe consequences on the phenotype than the individual absence of each of them. Induction of another CCR (CCR2, Atlg80820) and another CAD (CAD1, At4g39330) does not compensate the absence of the main CCR and CAD activities. This lack of CCR and CAD activities not only impacts lignification, but also severely affects the development of the plants. These consequences must be carefully considered when trying to reduce the lignin content of plants in order to facilitate the lignocellulose-to-bioethanol conversion process.
基金supported by the Office of Science (BER),U.S. Department of Energy grant DE-FG02-07ER64458 (Wilfred Vermerris and Scott E. Sattler)additional funding from USDA-ARS,CRIS project 5440-21220-032-00D (S.E.S,Deanna L. Funnell-Harris.)+2 种基金USDA AFRI grant number 2011-67009-30026 (S.E.S,D.L.F.H.)USDA Biomass Research and Development Initiative grant number 2011-10006-30358 (W.V.)the U.S. DOE’s International Affairs under award number DE-PI0000031 from the U.S. DOE’s Office of Energy Efficiency and Renewable Energy,Bioenergy Technologies Office (W.V)
文摘The presence of lignin reduces the quality of lignocellulosic biomass for forage materials and feedstock for biofuels. In C4 grasses, the brown midrib phenotype has been linked to mutations to genes in the monolignol biosynthesis pathway. For example, the Bmr6 gene in sorghum (Sorghum bicolor) has been previously shown to encode cinnamyl alcohol dehydrogenase (CAD), which catalyzes the final step of the monolignol biosynthesis pathway. Mutations in this gene have been shown to reduce the abundance of lignin, enhance digestibility, and improve saccharification efficiencies and ethanol yields. Nine sorghum lines harboring five different bmr6 alleles were identified in an EMS-mutagenized TILLING population. DNA sequencing of Bmr6 revealed that the majority of the mutations impacted evolutionarily conserved amino acids while three-dimensional structural modeling predicted that all of these alleles interfered with the enzyme's ability to bind with its NAPPH cofactor. All of the new alleles reduced in vitro CAD activity levels and enhanced glucose yields following saccharification. Further, many of these lines were associated with higher reductions in acid detergent lignin compared to lines harboring the previously characterized bmr6-ref allele. These bmr6 lines represent new breeding tools for manipulating biomass composition to enhance forage and feedstock quality.