OBJECTIVE: To evaluate the effects of aloin, cinnamic acid and 15 other kinds of natural chemicals on the activity of tyrosinase, in order to provide lightening agents in the treatment of hyperpigmentation disorders a...OBJECTIVE: To evaluate the effects of aloin, cinnamic acid and 15 other kinds of natural chemicals on the activity of tyrosinase, in order to provide lightening agents in the treatment of hyperpigmentation disorders and cosmetic additives. METHODS: Tyrosinase activity was estimated by measuring the oxidation rate of L-dopa. Inhibition of the enzyme was deduced according to the Lineweaver-Burk plots compared to the control. RESULTS: Cadabine, paeonal, farrerol, evodin, cinnamic acid, aloin and sophorcarpidine had different levels of inhibition of tyrosinase. The inhibitory rates of cinnamic acid (2 mmol/L, 0.5 mmol/L), aloin (2 mmol/L) and the rest were significantly higher than that of hydroquinone (0.5 mmol/L) (P展开更多
Plants synthesize the osmoprotectant glycine betaine (GB) via choline→betaine aldehyde→glycine be- taine[1]. Two enzymes are involved in the pathway choline monooxygenase (CMO) and betaine aldehyde dehydrogenase (BA...Plants synthesize the osmoprotectant glycine betaine (GB) via choline→betaine aldehyde→glycine be- taine[1]. Two enzymes are involved in the pathway choline monooxygenase (CMO) and betaine aldehyde dehydrogenase (BADH). A full length CMO cDNA (1,643bp) was cloned from Amaranthus tricolor. The open reading frame encoded a 442-amino acid polypeptide, which showed 69% identity with CMOs in Spina- cia oleracea L. and Beta vulgaris L. DNA gel blot analysis indicated the presence of one copy of CMO gene in the A. tricolor genome. The expressions of CMO and BADH proteins in A.tricolor leaves significantly increased under salinization, drought and heat stress (42℃), as determined by immunoblot analysis, but did not respond to cold stress (4℃), or exogenous ABA application. The increase of GB content in leaves was parallel to CMO and BADH contents.展开更多
Betaine is a very effective osmoprotectant found in many organisms. In high plants, betaine is synthesized by oxidation of choline in two sequential steps: choline-->betaine aldehyde-->betaine. The first step is...Betaine is a very effective osmoprotectant found in many organisms. In high plants, betaine is synthesized by oxidation of choline in two sequential steps: choline-->betaine aldehyde-->betaine. The first step is catalyzed by choline monooxygenase (CMO). In this study, the full-length CMO cDNA (1 820 bp) was cloned from halophyte Suaeda liaotungensis Kitag by RT-PCR and RACE. It included a 123 bp 5' UTR, a 368 bp 3' UTR and a 1 329 bp open reading frame encoding a 442-amino-acid polypeptide with 77%, 72% and 74% sequence identity compared to CMOs from spinach, sugar beet and Atriplex hortensis, respectively. The CMO open reading frame (ORF) was cloned and the plant expression vector pBI121-CMO was constructed. It was transferred into tobacco ( Nicotiana tabacum L. ev. 89) via Agrobacterium mediation. PCR and Southern blotting analysis showed that the CMO gene was integrated into tobacco genome. Transgenic tobacco plants contained higher amount of betaine than that of control plants and were able to survive on MS medium containing 250 mmol/L NaCl. Relative electronic conductivity demonstrated less membrane damage in transgenic plants as in the wild type.展开更多
The coupled effects of nitrogen source and methane monooxygenase（MMO） on the growth and poly-β-hydroxybutyrate（PHB） accumulation capacity of methanotrophs were explored.The ammonia-supplied methanotrophs expressi...The coupled effects of nitrogen source and methane monooxygenase（MMO） on the growth and poly-β-hydroxybutyrate（PHB） accumulation capacity of methanotrophs were explored.The ammonia-supplied methanotrophs expressing soluble MMO（s MMO） grew at the highest rate, while N2-fixing bacteria expressing particulate MMO（p MMO） grew at the lowest rate. Further study showed that more hydroxylamine and nitrite was formed by ammonia-supplied bacteria containing p MMO, which might cause their slightly lower growth rate. The highest PHB content（51.0%） was obtained under nitrogen-limiting conditions with the inoculation of nitrate-supplied bacteria containing p MMO. Ammoniasupplied bacteria also accumulated a higher content of PHB（45.2%） with the expression of p MMO, while N2-fixing bacteria containing p MMO only showed low PHB production capacity（32.1%）. The maximal PHB contents of bacteria expressing s MMO were low, with no significant change under different nitrogen source conditions. The low MMO activity,low cell growth rate and low PHB production capacity of methanotrophs continuously cultivated with N2 with the expression of p MMO were greatly improved in the cyclic NO3-N2 cultivation regime, indicating that long-term deficiency of nitrogen sources was detrimental to the activity of methanotrophs expressing pMMO.展开更多
Heavy metal pollution affects soil ecological function.Biochar and compost can effectively remediate heavy metals and increase soil nutrients.The effects and mechanisms of biochar and compost amendments on soil nitrog...Heavy metal pollution affects soil ecological function.Biochar and compost can effectively remediate heavy metals and increase soil nutrients.The effects and mechanisms of biochar and compost amendments on soil nitrogen cycle function in heavy-metal contaminated soils are not fully understood.This study examined how biochar,compost,and their integrated use affected ammonia-oxidizing microorganisms in heavy metal polluted soil.Quantitative PCR was used to determine the abundance of ammonia-oxidizing archaea(AOA)and bacteria(AOB).Ammonia monooxygenase(AMO)activity was evaluated by the enzymelinked immunosorbent assay.Results showed that compost rather than biochar improved nitrogen conversion in soil.Biochar,compost,or their integrated application significantly reduced the effective Zn and Cd speciation.Adding compost obviously increased As and Cu effective speciation,bacterial 16 S rRNA abundance,and AMO activity.AOB,stimulated by compost addition,was significantly more abundant than AOA throughout remediation.Correlation analysis showed that AOB abundance positively correlated with NO_(3)^(-)-N(r=0.830,P<0.01),and that AMO activity had significant correlation with EC(r=-0.908,P<0.01)and water-soluble carbon(r=-0.868,P<0.01).Those seem to be the most vital factors affecting AOB community and their function in heavy metal-polluted soil remediated by biochar and compost.展开更多
Glycyrrhiza uralensis is frequently used in traditional Chinese medicine. This plant contains a large amount of effective constituents, including triterpenoids and flavonoids. Among them, glycyrrhizin is believed to b...Glycyrrhiza uralensis is frequently used in traditional Chinese medicine. This plant contains a large amount of effective constituents, including triterpenoids and flavonoids. Among them, glycyrrhizin is believed to be the marker compound to evaluate the quality of G. uralensis based on Chinese Pharmacopoeia. Many studies showed that glycyrrhizin possesses various pharmacological activities, such as antibacterial, antiviral, antitumor, anti-inflammatory, and immune-stimulating activities. In this paper, we summarized the cloning, characterization, expression, and polymorphism analysis of several functional genes involved in glycyrrhizin biosynthesis in G. uralensis.展开更多
The cytochrome P450 monooxygenase （cytochrome P450） gene superfamily comprises many genes that may be involved in the biotransformations of pesticides and other xenobiotics. To date, very little is known about cytoc...The cytochrome P450 monooxygenase （cytochrome P450） gene superfamily comprises many genes that may be involved in the biotransformations of pesticides and other xenobiotics. To date, very little is known about cytochrome P450 genes in the oriental migratory locust, Locusta migratoria manilensis. In this study, we carried out a genomewide analysis ofcytochrome P450 genes of the locust to identify putative cytochrome P450 genes and characterize their expression responses to insecticide exposures. We identified 15 cytochrome P450-1ike genes from a locust expressed sequence tag database （LocustDB）. Reverse transcription polymerase chain reaction （RT-PCR） analysis showed that most cytochrome P450-1ike genes displayed different tissue and developmental stage expression patterns. However, most of them were predominantly expressed in the midgnt, gastric caeca, fatbodies, and/or hindgut. Biochemical analysis showed that cytochrome P450 was differentially affected by three different insecticides. Deltamethrin caused significant inductions in 12 h at LD30 （dose to kill 30% of the tested individuals） in the nymphs, whereas malathion and carbaryl did not have significant effect on cytochrome P450 enzyme activity. Further RT-PCR analysis Showed significant increases of transcriptions of several cytochrome P450 genes in deltamethrin-treated locusts. Thus, the increased cytochrome P450 enzyme activity is likely due to increased transcriptions of multiple cytochrome P450 genes in response to deltamethrin exposure. These results are expected to help us better understand the interactions between insecticides and major detoxification enzymes, and possible changes of the susceptibility to other insecticides in deltamethrin-treated insects at various molecular levels.展开更多
Cell-free expression systems have emerged as a versatile and powerful platform for metabolic engineering,biosynthesis and synthetic biology studies.Nevertheless,successful examples of the synthesis of complex natural ...Cell-free expression systems have emerged as a versatile and powerful platform for metabolic engineering,biosynthesis and synthetic biology studies.Nevertheless,successful examples of the synthesis of complex natural products using this system are still limited.Bicyclomycin,a structurally unique and complex diketopiperazine alkaloid,is a clinically promising antibiotic that selectively inhibits the transcription termination factor Rho.Here,we established a modular cell-free expression system with cascade catalysis for the biosynthesis of bicyclomycin from a chemically synthesized cyclodipeptide.The six cell-free expressed biosynthetic enzymes,including five iron-andα-ketoglutarate-dependent dioxygenases and one cytochrome P450 monooxygenase,were active in converting their substrates to the corresponding products.The co-expressed enzymes in the cell-free module were able to complete the related partial pathway.In vitro biosynthesis of bicyclomycin was also achieved by reconstituting the entire biosynthetic pathways(i.e.,six enzymes)using the modular cell-free expression system.This study demonstrates that the modular cell-free expression system can be used as a robust and promising platformforthe biosynthesis of complex antibiotics.展开更多
Serotonin deficiency in major depressive disorder(MDD)has formed the basis of antidepressant drug development and was originally attributed to induction of the major tryptophan(Trp)-degrading enzyme,liver Trp 2,3-diox...Serotonin deficiency in major depressive disorder(MDD)has formed the basis of antidepressant drug development and was originally attributed to induction of the major tryptophan(Trp)-degrading enzyme,liver Trp 2,3-dioxygenase(TDO),by cortisol,leading to decreased Trp availability to the brain for serotonin synthesis.Subsequently,the serotonin deficiency was proposed to involve induction of the extrahepatic Trp-degrading enzyme indoleamine 2,3-dioxygenase(IDO)by proinflammatory cytokines,with inflammation being the underlying cause.Recent evidence,however,challenges this latter concept,as not all MDD patients are immune-activated and,when present,inflammation is mild and/or transient.A wide range of antidepressant drugs inhibit the activity of liver TDO and bind specifically to the enzyme,but not to IDO.IDO induction is not a major event in MDD,but,when it occurs,its metabolic consequences may be masked and overridden by upregulation of kynurenine monooxygenase(KMO),the gateway to production of modulators of immune and neuronal functions.KMO appears to be activated in MDD by certain proinflammatory cytokines and antidepressants with anti-inflammatory properties may block this activation.We demonstrate the ability of the antidepressant ketamine to dock(bind)to KMO.The pathophysiology of MDD may be underpinned by both the serotonin deficiency and glutamatergic activation mediated respectively by TDO induction and N-methyl-D-aspartate receptor activation.Inhibition of TDO and KMO should be the focus of MDD pharmacotherapy.展开更多
Dioxygen activations constitute one of core issues in copper-dependent metalloenzymes. Upon O_(2) activation, copper-dependent metalloenzymes such as particulate methane monooxygenases(pM MOs), lytic polysaccharide mo...Dioxygen activations constitute one of core issues in copper-dependent metalloenzymes. Upon O_(2) activation, copper-dependent metalloenzymes such as particulate methane monooxygenases(pM MOs), lytic polysaccharide monooxygenases(LPMOs) and binuclear copper enzymes PHM and DβM, are able to perform various challenging C–H bond activations. Meanwhile, various copper-oxygen core containing complexes have been synthetized to mimic the active species of metalloenzymes. Dioxygen activation by mononuclear copper active site may generate various copper-oxygen intermediates, including Cu(Ⅱ)-superoxo, Cu(Ⅱ)-hydroperoxo, Cu(Ⅱ)-oxyl as well as the Cu(Ⅲ)-hydroxide species. Intriguingly, all these species have been invoked as the potential active intermediates for C–H/O–H activations in either biological or synthetic systems. Due to the poor understanding on reactivities of copper-oxygen complex, the nature of active species in both biological and synthetic systems are highly controversial. In this account, we will compare the reactivities of various mononuclear copper-oxygen species between biological systems and the synthetic systems. The present study is expected to provide the consistent understanding on reactivities of various copper-oxygen active species in both biological and synthetic systems.展开更多
This study evaluates the toxicological and biochemical response of two-spotted spider mites to clofentezine selection pressure. The mortality rate of Tetranychus urticae in adult females depends on increased clofentez...This study evaluates the toxicological and biochemical response of two-spotted spider mites to clofentezine selection pressure. The mortality rate of Tetranychus urticae in adult females depends on increased clofentezine concentration and clofentezine was found to be effective against females. The resistance rate of the CUM strain selected 12 times once per generation with clofentezine was increased from 1.28- to 105.27-fold. The interaction of some synergists with clofentezine was analyzed in the clofentezine-resistant CLOF 12 strain. Synergists had no effect on clofentezine toxicity. The clofentezine-resistant CLOF 12 strain showed resistance against chlorpyrifos, abamectin, propargite, fenpyroximate and amitraz. The modes of inheritance of resistance to clofentezine were found to be incompletely dominant and not sex-linked. Esterase enzyme activity was detected both by gel electrophoresis and microplate reader methods, while glutathione S-transferase （GST） and monooxygenase （P450） activity were detected only by the microplate reader method. During the selection period the esterase, the GST and the P450 enzymes activities were raised from 7.69, 7.09 and 0.003 3 to 18.40, 13.11 and 0.003 7 milli-optical density/rain/rag proteins, respectively. An increase was observed in the band intensity of esterases and esterase enzymes may play a role in clofentezine resistance in T. urticae.展开更多
文摘OBJECTIVE: To evaluate the effects of aloin, cinnamic acid and 15 other kinds of natural chemicals on the activity of tyrosinase, in order to provide lightening agents in the treatment of hyperpigmentation disorders and cosmetic additives. METHODS: Tyrosinase activity was estimated by measuring the oxidation rate of L-dopa. Inhibition of the enzyme was deduced according to the Lineweaver-Burk plots compared to the control. RESULTS: Cadabine, paeonal, farrerol, evodin, cinnamic acid, aloin and sophorcarpidine had different levels of inhibition of tyrosinase. The inhibitory rates of cinnamic acid (2 mmol/L, 0.5 mmol/L), aloin (2 mmol/L) and the rest were significantly higher than that of hydroquinone (0.5 mmol/L) (P
文摘Plants synthesize the osmoprotectant glycine betaine (GB) via choline→betaine aldehyde→glycine be- taine[1]. Two enzymes are involved in the pathway choline monooxygenase (CMO) and betaine aldehyde dehydrogenase (BADH). A full length CMO cDNA (1,643bp) was cloned from Amaranthus tricolor. The open reading frame encoded a 442-amino acid polypeptide, which showed 69% identity with CMOs in Spina- cia oleracea L. and Beta vulgaris L. DNA gel blot analysis indicated the presence of one copy of CMO gene in the A. tricolor genome. The expressions of CMO and BADH proteins in A.tricolor leaves significantly increased under salinization, drought and heat stress (42℃), as determined by immunoblot analysis, but did not respond to cold stress (4℃), or exogenous ABA application. The increase of GB content in leaves was parallel to CMO and BADH contents.
文摘Betaine is a very effective osmoprotectant found in many organisms. In high plants, betaine is synthesized by oxidation of choline in two sequential steps: choline-->betaine aldehyde-->betaine. The first step is catalyzed by choline monooxygenase (CMO). In this study, the full-length CMO cDNA (1 820 bp) was cloned from halophyte Suaeda liaotungensis Kitag by RT-PCR and RACE. It included a 123 bp 5' UTR, a 368 bp 3' UTR and a 1 329 bp open reading frame encoding a 442-amino-acid polypeptide with 77%, 72% and 74% sequence identity compared to CMOs from spinach, sugar beet and Atriplex hortensis, respectively. The CMO open reading frame (ORF) was cloned and the plant expression vector pBI121-CMO was constructed. It was transferred into tobacco ( Nicotiana tabacum L. ev. 89) via Agrobacterium mediation. PCR and Southern blotting analysis showed that the CMO gene was integrated into tobacco genome. Transgenic tobacco plants contained higher amount of betaine than that of control plants and were able to survive on MS medium containing 250 mmol/L NaCl. Relative electronic conductivity demonstrated less membrane damage in transgenic plants as in the wild type.
基金supported by the National Key Scientific and Technology Project for Water Pollution Treatment of China(No.2012ZX07202006)the National Natural Science Foundation of China(No.21477014)
文摘The coupled effects of nitrogen source and methane monooxygenase（MMO） on the growth and poly-β-hydroxybutyrate（PHB） accumulation capacity of methanotrophs were explored.The ammonia-supplied methanotrophs expressing soluble MMO（s MMO） grew at the highest rate, while N2-fixing bacteria expressing particulate MMO（p MMO） grew at the lowest rate. Further study showed that more hydroxylamine and nitrite was formed by ammonia-supplied bacteria containing p MMO, which might cause their slightly lower growth rate. The highest PHB content（51.0%） was obtained under nitrogen-limiting conditions with the inoculation of nitrate-supplied bacteria containing p MMO. Ammoniasupplied bacteria also accumulated a higher content of PHB（45.2%） with the expression of p MMO, while N2-fixing bacteria containing p MMO only showed low PHB production capacity（32.1%）. The maximal PHB contents of bacteria expressing s MMO were low, with no significant change under different nitrogen source conditions. The low MMO activity,low cell growth rate and low PHB production capacity of methanotrophs continuously cultivated with N2 with the expression of p MMO were greatly improved in the cyclic NO3-N2 cultivation regime, indicating that long-term deficiency of nitrogen sources was detrimental to the activity of methanotrophs expressing pMMO.
基金supported by the Hunan Provincial Key Research and Development Project(Nos.2019WK2031 and 2017SK2351)the National Natural Science Foundation of China(No.51408219)+1 种基金the Natural Science Foundation of Hu-nan Province(No.2020JJ5259)the Outstanding Youth Fund Project of the Hunan Education Department(No.18B094)。
文摘Heavy metal pollution affects soil ecological function.Biochar and compost can effectively remediate heavy metals and increase soil nutrients.The effects and mechanisms of biochar and compost amendments on soil nitrogen cycle function in heavy-metal contaminated soils are not fully understood.This study examined how biochar,compost,and their integrated use affected ammonia-oxidizing microorganisms in heavy metal polluted soil.Quantitative PCR was used to determine the abundance of ammonia-oxidizing archaea(AOA)and bacteria(AOB).Ammonia monooxygenase(AMO)activity was evaluated by the enzymelinked immunosorbent assay.Results showed that compost rather than biochar improved nitrogen conversion in soil.Biochar,compost,or their integrated application significantly reduced the effective Zn and Cd speciation.Adding compost obviously increased As and Cu effective speciation,bacterial 16 S rRNA abundance,and AMO activity.AOB,stimulated by compost addition,was significantly more abundant than AOA throughout remediation.Correlation analysis showed that AOB abundance positively correlated with NO_(3)^(-)-N(r=0.830,P<0.01),and that AMO activity had significant correlation with EC(r=-0.908,P<0.01)and water-soluble carbon(r=-0.868,P<0.01).Those seem to be the most vital factors affecting AOB community and their function in heavy metal-polluted soil remediated by biochar and compost.
基金Beijing Project for Young Talents(YETP0819)National Science Foundation of China(81503181)
文摘Glycyrrhiza uralensis is frequently used in traditional Chinese medicine. This plant contains a large amount of effective constituents, including triterpenoids and flavonoids. Among them, glycyrrhizin is believed to be the marker compound to evaluate the quality of G. uralensis based on Chinese Pharmacopoeia. Many studies showed that glycyrrhizin possesses various pharmacological activities, such as antibacterial, antiviral, antitumor, anti-inflammatory, and immune-stimulating activities. In this paper, we summarized the cloning, characterization, expression, and polymorphism analysis of several functional genes involved in glycyrrhizin biosynthesis in G. uralensis.
文摘The cytochrome P450 monooxygenase （cytochrome P450） gene superfamily comprises many genes that may be involved in the biotransformations of pesticides and other xenobiotics. To date, very little is known about cytochrome P450 genes in the oriental migratory locust, Locusta migratoria manilensis. In this study, we carried out a genomewide analysis ofcytochrome P450 genes of the locust to identify putative cytochrome P450 genes and characterize their expression responses to insecticide exposures. We identified 15 cytochrome P450-1ike genes from a locust expressed sequence tag database （LocustDB）. Reverse transcription polymerase chain reaction （RT-PCR） analysis showed that most cytochrome P450-1ike genes displayed different tissue and developmental stage expression patterns. However, most of them were predominantly expressed in the midgnt, gastric caeca, fatbodies, and/or hindgut. Biochemical analysis showed that cytochrome P450 was differentially affected by three different insecticides. Deltamethrin caused significant inductions in 12 h at LD30 （dose to kill 30% of the tested individuals） in the nymphs, whereas malathion and carbaryl did not have significant effect on cytochrome P450 enzyme activity. Further RT-PCR analysis Showed significant increases of transcriptions of several cytochrome P450 genes in deltamethrin-treated locusts. Thus, the increased cytochrome P450 enzyme activity is likely due to increased transcriptions of multiple cytochrome P450 genes in response to deltamethrin exposure. These results are expected to help us better understand the interactions between insecticides and major detoxification enzymes, and possible changes of the susceptibility to other insecticides in deltamethrin-treated insects at various molecular levels.
基金supported in part by grants from the National Key Research and Development Program of China(2022YFC2303100)the National Natural Science Foundation of China(22207117).
文摘Cell-free expression systems have emerged as a versatile and powerful platform for metabolic engineering,biosynthesis and synthetic biology studies.Nevertheless,successful examples of the synthesis of complex natural products using this system are still limited.Bicyclomycin,a structurally unique and complex diketopiperazine alkaloid,is a clinically promising antibiotic that selectively inhibits the transcription termination factor Rho.Here,we established a modular cell-free expression system with cascade catalysis for the biosynthesis of bicyclomycin from a chemically synthesized cyclodipeptide.The six cell-free expressed biosynthetic enzymes,including five iron-andα-ketoglutarate-dependent dioxygenases and one cytochrome P450 monooxygenase,were active in converting their substrates to the corresponding products.The co-expressed enzymes in the cell-free module were able to complete the related partial pathway.In vitro biosynthesis of bicyclomycin was also achieved by reconstituting the entire biosynthetic pathways(i.e.,six enzymes)using the modular cell-free expression system.This study demonstrates that the modular cell-free expression system can be used as a robust and promising platformforthe biosynthesis of complex antibiotics.
文摘Serotonin deficiency in major depressive disorder(MDD)has formed the basis of antidepressant drug development and was originally attributed to induction of the major tryptophan(Trp)-degrading enzyme,liver Trp 2,3-dioxygenase(TDO),by cortisol,leading to decreased Trp availability to the brain for serotonin synthesis.Subsequently,the serotonin deficiency was proposed to involve induction of the extrahepatic Trp-degrading enzyme indoleamine 2,3-dioxygenase(IDO)by proinflammatory cytokines,with inflammation being the underlying cause.Recent evidence,however,challenges this latter concept,as not all MDD patients are immune-activated and,when present,inflammation is mild and/or transient.A wide range of antidepressant drugs inhibit the activity of liver TDO and bind specifically to the enzyme,but not to IDO.IDO induction is not a major event in MDD,but,when it occurs,its metabolic consequences may be masked and overridden by upregulation of kynurenine monooxygenase(KMO),the gateway to production of modulators of immune and neuronal functions.KMO appears to be activated in MDD by certain proinflammatory cytokines and antidepressants with anti-inflammatory properties may block this activation.We demonstrate the ability of the antidepressant ketamine to dock(bind)to KMO.The pathophysiology of MDD may be underpinned by both the serotonin deficiency and glutamatergic activation mediated respectively by TDO induction and N-methyl-D-aspartate receptor activation.Inhibition of TDO and KMO should be the focus of MDD pharmacotherapy.
文摘Dioxygen activations constitute one of core issues in copper-dependent metalloenzymes. Upon O_(2) activation, copper-dependent metalloenzymes such as particulate methane monooxygenases(pM MOs), lytic polysaccharide monooxygenases(LPMOs) and binuclear copper enzymes PHM and DβM, are able to perform various challenging C–H bond activations. Meanwhile, various copper-oxygen core containing complexes have been synthetized to mimic the active species of metalloenzymes. Dioxygen activation by mononuclear copper active site may generate various copper-oxygen intermediates, including Cu(Ⅱ)-superoxo, Cu(Ⅱ)-hydroperoxo, Cu(Ⅱ)-oxyl as well as the Cu(Ⅲ)-hydroxide species. Intriguingly, all these species have been invoked as the potential active intermediates for C–H/O–H activations in either biological or synthetic systems. Due to the poor understanding on reactivities of copper-oxygen complex, the nature of active species in both biological and synthetic systems are highly controversial. In this account, we will compare the reactivities of various mononuclear copper-oxygen species between biological systems and the synthetic systems. The present study is expected to provide the consistent understanding on reactivities of various copper-oxygen active species in both biological and synthetic systems.
文摘This study evaluates the toxicological and biochemical response of two-spotted spider mites to clofentezine selection pressure. The mortality rate of Tetranychus urticae in adult females depends on increased clofentezine concentration and clofentezine was found to be effective against females. The resistance rate of the CUM strain selected 12 times once per generation with clofentezine was increased from 1.28- to 105.27-fold. The interaction of some synergists with clofentezine was analyzed in the clofentezine-resistant CLOF 12 strain. Synergists had no effect on clofentezine toxicity. The clofentezine-resistant CLOF 12 strain showed resistance against chlorpyrifos, abamectin, propargite, fenpyroximate and amitraz. The modes of inheritance of resistance to clofentezine were found to be incompletely dominant and not sex-linked. Esterase enzyme activity was detected both by gel electrophoresis and microplate reader methods, while glutathione S-transferase （GST） and monooxygenase （P450） activity were detected only by the microplate reader method. During the selection period the esterase, the GST and the P450 enzymes activities were raised from 7.69, 7.09 and 0.003 3 to 18.40, 13.11 and 0.003 7 milli-optical density/rain/rag proteins, respectively. An increase was observed in the band intensity of esterases and esterase enzymes may play a role in clofentezine resistance in T. urticae.
