L-cysteine desulfhydrase(CD)plays an impor-tant role in L-cysteine decomposition.To identify the CD gene in Pseudomonas sp.TS1138 and investigate its effect on the L-cysteine biosynthetic pathway,the CD gene was clone...L-cysteine desulfhydrase(CD)plays an impor-tant role in L-cysteine decomposition.To identify the CD gene in Pseudomonas sp.TS1138 and investigate its effect on the L-cysteine biosynthetic pathway,the CD gene was cloned from Pseudomonas sp.TS1138 by polymerase chain reaction(PCR)method.The nucleotide sequence of CD gene was determined to be 1,215 bp,and its homology with other sequences encoding CD was analyzed.Then the CD gene was subcloned into pET-21a(+)vector and expressed in Escherichia coli(E.coli)by isopropyl-β-D-thiogalacto-pyranoside(IPTG)inducement.The recombinant CD was purified by Ni-NTA His-Bind resin,and its activity was identified by the CD activity staining.The enzymatic proper-ties of the recombinant CD were characterized and its critical role involved in the L-cysteine biosynthetic pathway was also discussed.展开更多
Cryopreservation is associated with the production of reactive oxygen species, which leads to lipid peroxidation of the sperm membrane and consequently a reduction in sperm motility and decreased fertility potential. ...Cryopreservation is associated with the production of reactive oxygen species, which leads to lipid peroxidation of the sperm membrane and consequently a reduction in sperm motility and decreased fertility potential. The aim of this study was to determine the optimal concentration of L-cysteine needed for cryopreservation of boar semen. Twelve boars provided semen of proven motility and morphology for this study. The semen was divided into four portions in which the lactose-egg yolk (LEY) extender used to resuspend the centrifuged sperm pellet was supplemented with various concentrations of L-cysteine to reach 0 mmol L^-1 (group Ⅰ, control), 5 mmol L^-1 (group Ⅱ), 10 mmol L^-1 (group Ⅲ) and 15 mmol L^-1 (group Ⅳ). Semen suspensions were loaded in straws (0.5 mL) and placed in a controlled-rate freezer. After cryopreservation, frozen semen samples were thawed and investigated for progressive motility, viability using SYBR-14/EthD-1 staining and acrosome integrity using FITC-PNA/EthD-1 staining. There was a significantly higher (P 〈 0.01) percentage of progressive motility, viability and acrosomal integrity in two L-cysteine-supplemented groups (group Ⅱ and group Ⅲ) compared with the control. There was a biphasic effect of L-cysteine, with the highest percentage of progressive motility, viability and acrosomal integrity in group Ⅲ. In conclusion, 5 or 10 mmol L^-1 was the optimum concentration of L-cysteine to be added to the LEY extender for improving the quality of frozen-thawed boar semen.展开更多
The film contained L-cysteine and gold nanoparticles were provided by self-assembled monolayers (SAMs) and potentiostatic electrodeposition technology on the gold electrode. Two methods were used to study the film: In...The film contained L-cysteine and gold nanoparticles were provided by self-assembled monolayers (SAMs) and potentiostatic electrodeposition technology on the gold electrode. Two methods were used to study the film: In the first, cyclic voltammetry (CV) was used to inspect the functional groups of the film and the same time hydroquinone was chosen to be a probe molecule in the based solution;secondly, based on analytical technology of scanning electrochemical microscopy (SECM), the heterogeneous rate constant (keff) between solid phase (the modified electrode) and liquid phase (K3Fe(CN)6) was obtained. As a result, the better binary catalysis of hydroquinone was demonstrated and the heterogeneous rate constant (keff) is the greater at 8 h for L-cysteine self-assembled monolayers (SAMs).展开更多
An effective L-cysteine-assisted synthetic route has been successfully developed to prepare copper gallium sulfide(CuGaS2) microspheres under solvothermal conditions with CuCI2.2HzO, GaC13 and L-cysteine as source m...An effective L-cysteine-assisted synthetic route has been successfully developed to prepare copper gallium sulfide(CuGaS2) microspheres under solvothermal conditions with CuCI2.2HzO, GaC13 and L-cysteine as source materials, in which L-cysteine was used as the sulfide source and complexing molecule. The experiments revealed that the synthesized sample was of a typical CuGaS2 tetragonal structure. Moreover, the prepared CuGaS2 crystals consisting of microspheres made up of nanoflakes, and the diameter of the nanoflakes was about 20 nm. Raman spectrum of the obtained CuGaS2 exhibits a high-intensity peak of the A1 mode at 306 cm^-1. Meanwhile, a possible growth mechanism was proposed based on the investigations.展开更多
The effect of L-cysteine of argentum nitrate gel on the wound process of experimental animals is investigated. The results show that the nanogel, being bacteriostatic, accelerates the course of inflammation phase resu...The effect of L-cysteine of argentum nitrate gel on the wound process of experimental animals is investigated. The results show that the nanogel, being bacteriostatic, accelerates the course of inflammation phase resulting in fast cleansing of the wound surface and stimulation of the granulation tissue formation. The main cell elements of this tissue—fibroplasts are characterized by the state of functional excitation. The time of wound healing with the use of L-cysteine of argentum nitrate gel was reduced by three days, and the index of wound repair acceleration was about 20%.展开更多
We previously showed that hydrogen sulfide(H2S)has a neuroprotective effect in the context of hypoxic ischemic brain injury in neonatal mice.However,the precise mechanism underlying the role of H2S in this situation r...We previously showed that hydrogen sulfide(H2S)has a neuroprotective effect in the context of hypoxic ischemic brain injury in neonatal mice.However,the precise mechanism underlying the role of H2S in this situation remains unclear.In this study,we used a neonatal mouse model of hypoxic ischemic brain injury and a lipopolysaccharide-stimulated BV2 cell model and found that treatment with L-cysteine,a H2S precursor,attenuated the cerebral infarction and cerebral atrophy induced by hypoxia and ischemia and increased the expression of miR-9-5p and cystathionineβsynthase(a major H2S synthetase in the brain)in the prefrontal cortex.We also found that an miR-9-5p inhibitor blocked the expression of cystathionineβsynthase in the prefrontal cortex in mice with brain injury caused by hypoxia and ischemia.Furthermore,miR-9-5p overexpression increased cystathionine-β-synthase and H2S expression in the injured prefrontal cortex of mice with hypoxic ischemic brain injury.L-cysteine decreased the expression of CXCL11,an miR-9-5p target gene,in the prefrontal cortex of the mouse model and in lipopolysaccharide-stimulated BV-2 cells and increased the levels of proinflammatory cytokines BNIP3,FSTL1,SOCS2 and SOCS5,while treatment with an miR-9-5p inhibitor reversed these changes.These findings suggest that H2S can reduce neuroinflammation in a neonatal mouse model of hypoxic ischemic brain injury through regulating the miR-9-5p/CXCL11 axis and restoringβ-synthase expression,thereby playing a role in reducing neuroinflammation in hypoxic ischemic brain injury.展开更多
Hydrophilic interaction liquid chromatography(HILIC) has been recognized as an effective strategy for glycopeptide enrichment. Hydrophilic materials pave the way to solve the limit of low enrichment capacity and poor ...Hydrophilic interaction liquid chromatography(HILIC) has been recognized as an effective strategy for glycopeptide enrichment. Hydrophilic materials pave the way to solve the limit of low enrichment capacity and poor selectivity. The present study is the first attempt to combine chitosan(CS) and L-cysteine(L-Cys) to design a novel hydrophilic material focusing on glycopeptide enrichment. CS containing a large number of hydrophilic amino and hydroxyl groups has unique chemical properties, which makes it a very attractive biomaterial for glycopeptide enrichment. The excellent hydrophilicity of zwitterionic molecule L-Cys inspires the idea of anchoring L-Cys onto CS to design a novel hydrophilic material(named as Fe_(3)O_(4)@CS@Au-L-Cys) for the capture of low abundance glycopeptides. To be specific, Au nanoparticles(Au NPs) was introduced into CS-coated Fe_(3)O_(4)via electrostatic interaction and served as bridges to anchor L-Cys onto the surface of CS through strong Au-S bond interaction. The prepared Fe_(3)O_(4)@CS@AuL-Cys exhibited strong affinity, low detection limit(0.5 fmol/μL HRP), high selectivity(HRP/BSA with a molar ratio of 1:1000) for glycopeptides. Moreover, successful application of glycopeptide enrichment in human serum and saliva by Fe_(3)O_(4)@CS@Au-L-Cys was achieved. A satisfactory data set indicates that Fe_(3)O_(4)@CS@Au-L-Cys has promising potential in the application of glycopeptide enrichment in real complex bio-samples and for related glycoproteome research.展开更多
The objective of the present study was to determine the effects of docosahexaenoic acid (DHA)-enriched hen egg yolks and L-cysteine supplementation on the qualities of the cryopreserved boar semen. A total of 15 eja...The objective of the present study was to determine the effects of docosahexaenoic acid (DHA)-enriched hen egg yolks and L-cysteine supplementation on the qualities of the cryopreserved boar semen. A total of 15 ejaculates from 5 Pietrain boars were divided into 4 groups according to the compositions of the freezing extenders used, that is, normal hen egg yolk (group Ⅰ), DHA-enriched hen egg yolk (group Ⅱ), normal hen egg yolk with 5 mmol L^-1 of cysteine supplementation (group Ⅲ) and DHA-enriched hen egg yolk with 5 mmol L1 of cysteine supplementation (group Ⅳ). The semen was cryopreserved using controlled rate freezer and was thawed at 50℃ for 12 s. Progressive motility, sperm viability, acrosome integrity and functional integrity of sperm plasma membrane of the post-thawed semen were evaluated. The supplementation of L-cysteine in the freezing extender alone (group Ⅲ) improved progressive motility (P 〈 0.05), and the supplementation of L-cysteine in combination with DHA-enriched hen egg yolk (group Ⅳ) improved both progressive motility (P 〈 0.05) and acrosome integrity (P 〈 0.01). The use of DHA-enriched hen egg yolk alone (group Ⅱ) did not enhance any of the post-thawed semen qualities (P 〉 0.05). In conclusion, the supplementation of antioxidant L-cysteine alone or in combination with DHA-enriched hen egg yolk significantly improved the post-thawed semen qualities, especially progressive motility and acrosome integrity.展开更多
L-Cysteine plays a prominent role in sulfur metabo- lism of plants. However, its role in root development is largely unknown. Here, we report that L-cysteine reduces primary root growth in a dosage-dependent manner. E...L-Cysteine plays a prominent role in sulfur metabo- lism of plants. However, its role in root development is largely unknown. Here, we report that L-cysteine reduces primary root growth in a dosage-dependent manner. Elevating cellular L-cysteine level by exposing Arabidopsis thaliana seedlings to high L-cysteine, buthionine sulphoximine, or O-acetylserine leads to altered auxin maximum in root tips, the expression of quiescent center cell marker as well as the decrease of the auxin carriers PIN1, PIN2, PIN3, and PIN7 of primary roots. We also show that high L-cysteine significantly reduces the protein level of two sets of stem cell specific transcription factors PLETHORA1/2 and SCR/SHR. However, L-cysteine does not downregulate the transcript level of PiNs, PLTs, or SCR/SHR, suggesting that an uncharacterized post-transcriptional mechanism may regulate the accumulation of PIN, PLT, and SCR/SHR proteins and auxin transport in the root tips. These results suggest that endogenous L-cysteine level acts to maintain root stem cell niche by regulating basal- and auxin-induced expression of PLT1/2 and 5CR/SHR. L-Cysteine may serve as a link between sulfate assimilation and auxin in regulating root growth.展开更多
The electrooxidation of L-cysteine(L-Cys) was studied using a benzoylferrocene(BF) modified multi-wall carbon nanotube paste electrode(BFCNPE) using cyclic voltammetry(CV),square wave voltammetry(SWV) and ch...The electrooxidation of L-cysteine(L-Cys) was studied using a benzoylferrocene(BF) modified multi-wall carbon nanotube paste electrode(BFCNPE) using cyclic voltammetry(CV),square wave voltammetry(SWV) and chronoamperometry(CHA).Under optimum pH in CV the oxidation of L-Cys occurs at a potential about 215 mV less positive than that at the surface of unmodified carbon paste electrode.The catalytic oxidation peak currents were dependent on the L-Cys concentration and a linear calibration curve was obtained in the range 0.7-350.0 mmol/L of L-Cys with SWV method.The detection limit(3s) was determined as 0.1 mmol/L.This method was also used for the determination of L-Cys in some real samples.展开更多
基金This work was supported by Grant no.30470053 from the National Natural Science Foundation of China and Grant no.05YFJZJC00900 from the Natural Science Foundation of Tianjin,China.
文摘L-cysteine desulfhydrase(CD)plays an impor-tant role in L-cysteine decomposition.To identify the CD gene in Pseudomonas sp.TS1138 and investigate its effect on the L-cysteine biosynthetic pathway,the CD gene was cloned from Pseudomonas sp.TS1138 by polymerase chain reaction(PCR)method.The nucleotide sequence of CD gene was determined to be 1,215 bp,and its homology with other sequences encoding CD was analyzed.Then the CD gene was subcloned into pET-21a(+)vector and expressed in Escherichia coli(E.coli)by isopropyl-β-D-thiogalacto-pyranoside(IPTG)inducement.The recombinant CD was purified by Ni-NTA His-Bind resin,and its activity was identified by the CD activity staining.The enzymatic proper-ties of the recombinant CD were characterized and its critical role involved in the L-cysteine biosynthetic pathway was also discussed.
文摘Cryopreservation is associated with the production of reactive oxygen species, which leads to lipid peroxidation of the sperm membrane and consequently a reduction in sperm motility and decreased fertility potential. The aim of this study was to determine the optimal concentration of L-cysteine needed for cryopreservation of boar semen. Twelve boars provided semen of proven motility and morphology for this study. The semen was divided into four portions in which the lactose-egg yolk (LEY) extender used to resuspend the centrifuged sperm pellet was supplemented with various concentrations of L-cysteine to reach 0 mmol L^-1 (group Ⅰ, control), 5 mmol L^-1 (group Ⅱ), 10 mmol L^-1 (group Ⅲ) and 15 mmol L^-1 (group Ⅳ). Semen suspensions were loaded in straws (0.5 mL) and placed in a controlled-rate freezer. After cryopreservation, frozen semen samples were thawed and investigated for progressive motility, viability using SYBR-14/EthD-1 staining and acrosome integrity using FITC-PNA/EthD-1 staining. There was a significantly higher (P 〈 0.01) percentage of progressive motility, viability and acrosomal integrity in two L-cysteine-supplemented groups (group Ⅱ and group Ⅲ) compared with the control. There was a biphasic effect of L-cysteine, with the highest percentage of progressive motility, viability and acrosomal integrity in group Ⅲ. In conclusion, 5 or 10 mmol L^-1 was the optimum concentration of L-cysteine to be added to the LEY extender for improving the quality of frozen-thawed boar semen.
文摘The film contained L-cysteine and gold nanoparticles were provided by self-assembled monolayers (SAMs) and potentiostatic electrodeposition technology on the gold electrode. Two methods were used to study the film: In the first, cyclic voltammetry (CV) was used to inspect the functional groups of the film and the same time hydroquinone was chosen to be a probe molecule in the based solution;secondly, based on analytical technology of scanning electrochemical microscopy (SECM), the heterogeneous rate constant (keff) between solid phase (the modified electrode) and liquid phase (K3Fe(CN)6) was obtained. As a result, the better binary catalysis of hydroquinone was demonstrated and the heterogeneous rate constant (keff) is the greater at 8 h for L-cysteine self-assembled monolayers (SAMs).
基金Supported by the National Natural Science Foundation of China(Nos.507872075 and 50972107)the Project of Key Scientific and Technological Innovations Team of Zhejiang Province,China(No.2009R50010)the Key Foundation of Natural Science Foundation of Zhejiang Province,China(No.Z4110347)
文摘An effective L-cysteine-assisted synthetic route has been successfully developed to prepare copper gallium sulfide(CuGaS2) microspheres under solvothermal conditions with CuCI2.2HzO, GaC13 and L-cysteine as source materials, in which L-cysteine was used as the sulfide source and complexing molecule. The experiments revealed that the synthesized sample was of a typical CuGaS2 tetragonal structure. Moreover, the prepared CuGaS2 crystals consisting of microspheres made up of nanoflakes, and the diameter of the nanoflakes was about 20 nm. Raman spectrum of the obtained CuGaS2 exhibits a high-intensity peak of the A1 mode at 306 cm^-1. Meanwhile, a possible growth mechanism was proposed based on the investigations.
文摘The effect of L-cysteine of argentum nitrate gel on the wound process of experimental animals is investigated. The results show that the nanogel, being bacteriostatic, accelerates the course of inflammation phase resulting in fast cleansing of the wound surface and stimulation of the granulation tissue formation. The main cell elements of this tissue—fibroplasts are characterized by the state of functional excitation. The time of wound healing with the use of L-cysteine of argentum nitrate gel was reduced by three days, and the index of wound repair acceleration was about 20%.
基金supported by the National Natural Science Foundation of China,Nos.82271327(to ZW),82072535(to ZW),81873768(to ZW),and 82001253(to TL).
文摘We previously showed that hydrogen sulfide(H2S)has a neuroprotective effect in the context of hypoxic ischemic brain injury in neonatal mice.However,the precise mechanism underlying the role of H2S in this situation remains unclear.In this study,we used a neonatal mouse model of hypoxic ischemic brain injury and a lipopolysaccharide-stimulated BV2 cell model and found that treatment with L-cysteine,a H2S precursor,attenuated the cerebral infarction and cerebral atrophy induced by hypoxia and ischemia and increased the expression of miR-9-5p and cystathionineβsynthase(a major H2S synthetase in the brain)in the prefrontal cortex.We also found that an miR-9-5p inhibitor blocked the expression of cystathionineβsynthase in the prefrontal cortex in mice with brain injury caused by hypoxia and ischemia.Furthermore,miR-9-5p overexpression increased cystathionine-β-synthase and H2S expression in the injured prefrontal cortex of mice with hypoxic ischemic brain injury.L-cysteine decreased the expression of CXCL11,an miR-9-5p target gene,in the prefrontal cortex of the mouse model and in lipopolysaccharide-stimulated BV-2 cells and increased the levels of proinflammatory cytokines BNIP3,FSTL1,SOCS2 and SOCS5,while treatment with an miR-9-5p inhibitor reversed these changes.These findings suggest that H2S can reduce neuroinflammation in a neonatal mouse model of hypoxic ischemic brain injury through regulating the miR-9-5p/CXCL11 axis and restoringβ-synthase expression,thereby playing a role in reducing neuroinflammation in hypoxic ischemic brain injury.
基金financially supported by Open Project of State Key Laboratory of Supramolecular Structure and Materials,Jilin University,China(No.sklssm2022012)the Fundamental Research Funds for the Central Universities,JLU,China。
文摘Hydrophilic interaction liquid chromatography(HILIC) has been recognized as an effective strategy for glycopeptide enrichment. Hydrophilic materials pave the way to solve the limit of low enrichment capacity and poor selectivity. The present study is the first attempt to combine chitosan(CS) and L-cysteine(L-Cys) to design a novel hydrophilic material focusing on glycopeptide enrichment. CS containing a large number of hydrophilic amino and hydroxyl groups has unique chemical properties, which makes it a very attractive biomaterial for glycopeptide enrichment. The excellent hydrophilicity of zwitterionic molecule L-Cys inspires the idea of anchoring L-Cys onto CS to design a novel hydrophilic material(named as Fe_(3)O_(4)@CS@Au-L-Cys) for the capture of low abundance glycopeptides. To be specific, Au nanoparticles(Au NPs) was introduced into CS-coated Fe_(3)O_(4)via electrostatic interaction and served as bridges to anchor L-Cys onto the surface of CS through strong Au-S bond interaction. The prepared Fe_(3)O_(4)@CS@AuL-Cys exhibited strong affinity, low detection limit(0.5 fmol/μL HRP), high selectivity(HRP/BSA with a molar ratio of 1:1000) for glycopeptides. Moreover, successful application of glycopeptide enrichment in human serum and saliva by Fe_(3)O_(4)@CS@Au-L-Cys was achieved. A satisfactory data set indicates that Fe_(3)O_(4)@CS@Au-L-Cys has promising potential in the application of glycopeptide enrichment in real complex bio-samples and for related glycoproteome research.
文摘The objective of the present study was to determine the effects of docosahexaenoic acid (DHA)-enriched hen egg yolks and L-cysteine supplementation on the qualities of the cryopreserved boar semen. A total of 15 ejaculates from 5 Pietrain boars were divided into 4 groups according to the compositions of the freezing extenders used, that is, normal hen egg yolk (group Ⅰ), DHA-enriched hen egg yolk (group Ⅱ), normal hen egg yolk with 5 mmol L^-1 of cysteine supplementation (group Ⅲ) and DHA-enriched hen egg yolk with 5 mmol L1 of cysteine supplementation (group Ⅳ). The semen was cryopreserved using controlled rate freezer and was thawed at 50℃ for 12 s. Progressive motility, sperm viability, acrosome integrity and functional integrity of sperm plasma membrane of the post-thawed semen were evaluated. The supplementation of L-cysteine in the freezing extender alone (group Ⅲ) improved progressive motility (P 〈 0.05), and the supplementation of L-cysteine in combination with DHA-enriched hen egg yolk (group Ⅳ) improved both progressive motility (P 〈 0.05) and acrosome integrity (P 〈 0.01). The use of DHA-enriched hen egg yolk alone (group Ⅱ) did not enhance any of the post-thawed semen qualities (P 〉 0.05). In conclusion, the supplementation of antioxidant L-cysteine alone or in combination with DHA-enriched hen egg yolk significantly improved the post-thawed semen qualities, especially progressive motility and acrosome integrity.
文摘L-Cysteine plays a prominent role in sulfur metabo- lism of plants. However, its role in root development is largely unknown. Here, we report that L-cysteine reduces primary root growth in a dosage-dependent manner. Elevating cellular L-cysteine level by exposing Arabidopsis thaliana seedlings to high L-cysteine, buthionine sulphoximine, or O-acetylserine leads to altered auxin maximum in root tips, the expression of quiescent center cell marker as well as the decrease of the auxin carriers PIN1, PIN2, PIN3, and PIN7 of primary roots. We also show that high L-cysteine significantly reduces the protein level of two sets of stem cell specific transcription factors PLETHORA1/2 and SCR/SHR. However, L-cysteine does not downregulate the transcript level of PiNs, PLTs, or SCR/SHR, suggesting that an uncharacterized post-transcriptional mechanism may regulate the accumulation of PIN, PLT, and SCR/SHR proteins and auxin transport in the root tips. These results suggest that endogenous L-cysteine level acts to maintain root stem cell niche by regulating basal- and auxin-induced expression of PLT1/2 and 5CR/SHR. L-Cysteine may serve as a link between sulfate assimilation and auxin in regulating root growth.
文摘The electrooxidation of L-cysteine(L-Cys) was studied using a benzoylferrocene(BF) modified multi-wall carbon nanotube paste electrode(BFCNPE) using cyclic voltammetry(CV),square wave voltammetry(SWV) and chronoamperometry(CHA).Under optimum pH in CV the oxidation of L-Cys occurs at a potential about 215 mV less positive than that at the surface of unmodified carbon paste electrode.The catalytic oxidation peak currents were dependent on the L-Cys concentration and a linear calibration curve was obtained in the range 0.7-350.0 mmol/L of L-Cys with SWV method.The detection limit(3s) was determined as 0.1 mmol/L.This method was also used for the determination of L-Cys in some real samples.