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代谢工程改造Corynebacterium glutamicum生产L-苹果酸 被引量:1
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作者 赵一 李天明 +3 位作者 刘金雷 王崇慧 仪宏 冯惠勇 《食品与发酵工业》 CAS CSCD 北大核心 2016年第12期13-19,共7页
以提高L-苹果酸的产量为目标,采用一次交换两次同源重组的方法,利用反向筛选标记,在敲除了丙酮酸醌氧化还原酶编码基因(pqo),丙酮酸脱氢酶编码基因(pdh)和乳酸脱氢酶编码基因(lldh)的C.glutamicumΔpqoΔpdhΔlldh(C.glutamicumΔPPL)... 以提高L-苹果酸的产量为目标,采用一次交换两次同源重组的方法,利用反向筛选标记,在敲除了丙酮酸醌氧化还原酶编码基因(pqo),丙酮酸脱氢酶编码基因(pdh)和乳酸脱氢酶编码基因(lldh)的C.glutamicumΔpqoΔpdhΔlldh(C.glutamicumΔPPL)基础上,无痕敲除了L-苹果酸积累支流代谢途径的2个关键酶基因:苹果酸醌氧化还原酶编码基因(mqo)和苹果酸酶编码基因(male),同时敲入了苹果酸分泌转运蛋白基因(transb),获得了产L-苹果酸的工程菌株;采用高效液相色谱法检测了工程菌株C.glutamicumΔPPLΔmqo::transbΔmale的发酵产物。实验结果表明:C.glutamicum ATCC 13032发酵后不积累L-苹果酸,而工程菌C.glutamicumΔPPLΔmqo::transbΔmale发酵48 h,积累了12.8 g/L的L-苹果酸,工程菌的糖酸转化率为33.18%,为利用C.glutamicum ATCC 13032发酵生产L-苹果酸提供了基础遗传资源。 展开更多
关键词 谷氨酸棒状杆菌(corynebacterium glutamicum) L-苹果酸 基因敲除 基因敲入
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奶牛临床型乳房炎Corynebacterium phoceense的分离鉴定及药敏试验 被引量:1
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作者 姜东凤 唐启伟 +3 位作者 肖艳飞 任晓丽 皇甫和平 石冬梅 《家畜生态学报》 北大核心 2021年第4期63-66,83,共5页
对太原市某奶牛场临床型乳房炎病牛进行病原菌的分离鉴定及药敏试验,以期了解主要的病原菌及其药物敏感性,为该场乳房炎的防控提供参考依据。本研究采用病原菌分离培养、16S rRNA基因的PCR扩增及序列分析、16联牛乳房炎病原菌核酸qPCR-... 对太原市某奶牛场临床型乳房炎病牛进行病原菌的分离鉴定及药敏试验,以期了解主要的病原菌及其药物敏感性,为该场乳房炎的防控提供参考依据。本研究采用病原菌分离培养、16S rRNA基因的PCR扩增及序列分析、16联牛乳房炎病原菌核酸qPCR-荧光探针法鉴定了该牧场4头患病牛的9份乳样,并用纸片法对分离菌株进行了药敏试验。结果表明,共分离出8株细菌,包括Corynebacterium phoceense 5株(62.5%)、气球菌1株、链球菌1株和嗜冷杆菌1株,这些菌属对环丙沙星、庆大霉素、阿莫西林拉维酸钾、利福平、多西环素、头孢噻呋均为高度敏感,而对氨曲南、磺胺异恶唑有一定耐药性;同时,通过TaqMan探针实时荧光PCR筛选出2种弱阳性菌属和6种疑似菌属,但是病原菌含量均不高,且大多为环境条件性致病菌。研究结果说明分离培养的方法不能全面地反映引起乳房炎的病原菌感染状况。所以在生产中须借助分子生物学方法进行综合检测分析。该试验也初步判定Corynebacterium phoceense可能是引起乳房炎的主要致病菌之一。 展开更多
关键词 奶牛乳房炎 corynebacterium phoceense 分离鉴定 药敏试验 16S rRNA基因检测 qPCR鉴定
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Increased L-arginine Production by Site-directed Mutagenesis of N-acetyl-L-glutamate Kinase and pro B Gene Deletion in Corynebacterium crenatum 被引量:5
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作者 ZHANG Bin WAN Fang +4 位作者 QIU Yu Lou CHEN Xue Lan TANG Li CHEN Jin Cong XIONG Yong Hua 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2015年第12期864-874,共11页
Objective In Corynebacterium crenatum,the adjacent D311 and D312 of N-acetyl-L-glutamate kinase(NAGK),as a key rate-limiting enzyme of L-arginine biosynthesis under substrate regulatory control by arginine,were initia... Objective In Corynebacterium crenatum,the adjacent D311 and D312 of N-acetyl-L-glutamate kinase(NAGK),as a key rate-limiting enzyme of L-arginine biosynthesis under substrate regulatory control by arginine,were initially replaced with two arginine residues to investigate the L-arginine feedback inhibition for NAGK.Methods NAGK enzyme expression was evaluated using a plasmid-based method.Homologous recombination was employed to eliminate the pro B.Results The IC50 and enzyme activity of NAGK M4,in which the D311 R and D312 R amino acid substitutions were combined with the previously reported E19 R and H26 E substitutions,were 3.7-fold and 14.6% higher,respectively,than those of the wild-type NAGK.NAGK M4 was successfully introduced into the C.crenatum MT genome without any genetic markers;the L-arginine yield of C.crenatum MT-M4 was 26.2% higher than that of C.crenatum MT.To further improve upon the L-arginine yield,we constructed the mutant C.crenatum MT-M4 ?pro B.The optimum concentration of L-proline was also investigated in order to determine its contribution to L-arginine yield.After L-proline was added to the medium at 10 mmol/L,the L-arginine yield reached 16.5 g/L after 108 h of shake-flask fermentation,approximately 70.1% higher than the yield attained using C.crenatum MT.Conclusion Feedback inhibition of L-arginine on NAGK in C.crenatum is clearly alleviated by the M4 mutation of NAGK,and deletion of the pro B in C.crenatum from MT to M4 results in a significant increase in arginine production. 展开更多
关键词 corynebacterium crenatum N-acetyl-L-glutamate kinase Site-directed mutagenesis L-ARGININE proB
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Desulfurization of dibenzothiophene by a newly isolated Corynebacterium sp. ZD-1 in aqueous phase 被引量:4
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作者 WANGMiao-dong LIWei WANGDa-hui SHIYao 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2004年第6期1011-1015,共5页
Sulfur emission through fuel combustion is a global problem because it is a major cause of acid rain. Crud oil contains many heterocyclic organic sulfur compounds, among which dibenzothiophene(DBT) and DBTs bearing al... Sulfur emission through fuel combustion is a global problem because it is a major cause of acid rain. Crud oil contains many heterocyclic organic sulfur compounds, among which dibenzothiophene(DBT) and DBTs bearing alkyl substitutions usually are representative compounds. A strain was isolated from refinery sludge and identified as Corynebacterium ZD-1. The behavior of DBT degradation by ZD-1 in aqueous phase was investigated. Corynebacterium ZD-1 could metabolize DBT to 2-hydroxybiphenyl(2-HBP) as the dead-end metabolite through a sulfur-specific pathway. In shake flask culture, ZD-1 had its maximal desulfurization activity in the late exponential growth phase and the specific production rate of 2-HBP was about 0.14(mmol·kg dry cell -1·min -1, mmol·KDC -1·min -1). Active resting cells for desulfurization should be prepared only in this period. 2-HBP inhibited the growth of strain ZD-1, the production of DBT degradation enzymes, and the activity of enzymes. Sulfate inhibited the production of dibenzothiophene(DBT) degradation enzymes but had no effect on the enzymes' activity. The production rates of 2-HBP at lower cell densities were higher and the maximum amount conversion of DBT to 2-HBP(0.067 mmol/L) after 8 h was gained at 9.2 g dry cell/L rather higher cell density. The results indicated that this newly isolated strain could be a promising biocatalyst for DBT desulfurization. 展开更多
关键词 BIODESULFURIZATION DIBENZOTHIOPHENE corynebacterium
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Two-stage pH Control Mode in Batch Fermentation of a Novel Bioflocculant from Corynebacterium Glutamicum 被引量:3
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作者 HENing WUXiao-jie DENGXu LUYing-hua LIQing-biao 《Chemical Research in Chinese Universities》 SCIE CAS CSCD 2004年第2期152-155,共4页
The effect of pH of the fermentation medium on cell growth and the production of a novel bioflocculant(named REA-11) by Corynebacterium glutamicum CCTCC M201005 were investigated. The maximum biomass(2.23 g/L) and fl... The effect of pH of the fermentation medium on cell growth and the production of a novel bioflocculant(named REA-11) by Corynebacterium glutamicum CCTCC M201005 were investigated. The maximum biomass(2.23 g/L) and flocculating activity(142.2 U/mL) were simultaneously obtained at the 14th hour when the pH value of the culture medium was maintained at 7.0 during the whole fermentation process. The production of REA-11 kept on a trend of increase till the later phase of fermentation process, which resulted in the ultimate flocculating activity of the culture broth to enhance to nearly 100 U/mL at pH 6.0. A two-stage pH control mode was adopted in REA-11 production in which the pH value of the culture medium was controlled at 7.0 during the first 14 h, then decreased to 6.0 that was maintained until the end of the fermentation process. With the two-stage pH control mode, the maximum flocculating activity reached 178.8 U/mL which was 30% higher than that obtained under the condition of pH 7.0 and the biomass enhanced about 15%. Compared with the fermentation process without pH control, REA-11 production and cell growth via the two-stage pH control mode increased 80% and 25%, respectively. 展开更多
关键词 corynebacterium glutamicum pH control BIOFLOCCULANT
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Expression and Characterization of ArgR, An Arginine Regulatory Protein in Corynebacterium crenatum 被引量:2
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作者 CHEN Xue Lan ZHANG Bin +6 位作者 TANG Li JIAO Hai Tao XU Heng Yi XU Feng XU Hong WEI Hua XIONG Yong Hua 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2014年第6期436-443,共8页
Objective Corynebacterium crenatum MT, a mutant from C. crenatum AS 1.542 with a lethal argR gene, exhibits high arginine production. To confirm the effect of ArgR on arginine biosynthesis in C. crenatum, an intact ar... Objective Corynebacterium crenatum MT, a mutant from C. crenatum AS 1.542 with a lethal argR gene, exhibits high arginine production. To confirm the effect of ArgR on arginine biosynthesis in C. crenatum, an intact argR gene from wild-type AS 1.542 was introduced into C. crenatum MT, resulting in C. crenatum MT. sp, and the changes of transcriptional levels of the arginine biosynthetic genes and arginine production were compared between the mutant strain and the recombinant strain. Methods Quantitative real-time polymerase chain reaction was employed to analyze the changes of the related genes at the transcriptional level, electrophoretic mobility shift assays were used to determine ArgR binding with the argCJBDF, argGH, and carAB promoter regions, and arginine production was determined with an automated amino acid analyzer. Results Arginine production assays showed a 69.9% reduction in arginine from 9.01±0.22 mg/mL in C. crenatum MT to 2.71±0.13 mg/mL (P〈0.05) in C. crenatum MT. sp. The argC, argB, argD, argF, argJ, argG, and carA genes were down-regulated significantly in C. crenatum MT. sp compared with those in its parental C. crenatum MT strain. The electrophoretic mobility shift assays showed that the promoter regions were directly bound to the ArgR protein. Conclusion The arginine biosynthetic genes in C crenatum are clearly controlled by the regulator ArgR, and intact ArgR in C. crenatum MT results in a significant descrease in production. negative arginine production. 展开更多
关键词 corynebacterium crenatum ArgR protein Arginine biosynthetic genes Real-time PCR ElectrophoretJc mobility shift assay
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稀土元素镧对Corynebacterium pekinense发酵赖氨酸的影响 被引量:4
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作者 张颖 《安徽农业科学》 CAS 2012年第10期5741-5742,5810,共3页
[目的]研究稀土元素镧对北京棒杆菌(Corynebacterium pekinense)的生长及产物合成的影响。[方法]通过全因子试验设计,分析了在北京棒杆菌的发酵培养基中添加不同配比稀土元素镧对其赖氨酸产量的影响。[结果]低剂量的稀土元素镧可以促进... [目的]研究稀土元素镧对北京棒杆菌(Corynebacterium pekinense)的生长及产物合成的影响。[方法]通过全因子试验设计,分析了在北京棒杆菌的发酵培养基中添加不同配比稀土元素镧对其赖氨酸产量的影响。[结果]低剂量的稀土元素镧可以促进北京棒杆菌生长并积累代谢产物赖氨酸,利用Matlab软件优化培养基中显著影响成分,得到稀土镧离子的最佳浓度为78 mg/L,葡萄糖为36 g/L,牛肉膏为4.5 g/L,蛋白胨为20 g/L。[结论]该研究初步确定了在北京棒杆菌培养中,La+的适宜添加量及该菌产生赖氨酸的最适培养基组成。 展开更多
关键词 稀土元素 谷氨酸棒杆菌 赖氨酸发酵
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Analysis of the arginine biosynthetic gene cluster argCJBDFR of Corynebacterium crenatum 被引量:1
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作者 Haitao Jiao Yong Yuan +1 位作者 Yonghua Xiong Xuelan Chen 《Journal of Biomedical Science and Engineering》 2011年第1期70-75,共6页
Objective: Corynebacterium crenatum AS1.542, a Gram-positive bacterium and indigenous nonpatho-genic corynebacteria, is widely exploited for the in-dustrial production of amino acids. The objective of this paper is to... Objective: Corynebacterium crenatum AS1.542, a Gram-positive bacterium and indigenous nonpatho-genic corynebacteria, is widely exploited for the in-dustrial production of amino acids. The objective of this paper is to clarify the genetic information of the arginine biosynthetic pathway, and further more contribute to the improvement of arginine produc-tion. Methods: Polymerase chain reaction (PCR) technology was employed for obtaining the arginine biosynthetic gene sequence, and softwares eg. Laser-gene, BPROM, RNAshapes were used for the analysis of obtained sequences. Results: Arginine biosynethetic gene cluster of C. crenatum, comprising argJ, argB, argD, argF, argR and part of argC, has been ampli-fied and sequenced. The gene order has been estab-lished as argCJBDFR, with a entire length of 6.08kb. Conclusion: An internal promoter was found in the upstream of argB gene, four argBDFR ORFs are lo-cated in a same transcription unit, and the tran-scripiton termination of argC gene is irrelevant with the rho-factor. Comparison with ornithine acetyl-transferase (coded by argJ gene) from C. glutamate, ornithine acetyltransferase from C. crenatum also belongs to the monofunctional enzymes. 展开更多
关键词 corynebacterium crenatum argCJBDFR SEQUENCE ORNITHINE ACETYLTRANSFERASE argR GENE
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细胞密度对Corynebacterium sp. NNC-3降解苯酚的影响
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作者 何熙璞 陈加辉 +4 位作者 刘鸿杰 张敏 梁锦添 刘婕 陈保善 《广西大学学报(自然科学版)》 CAS 北大核心 2013年第3期663-667,共5页
以Corynebacterium sp.NNC-3为受试菌株,就初始细胞密度对苯酚降解反应的影响进行了研究。结果表明,初始苯酚浓度为100~600 mg/L,初始细胞密度在0.05~0.30(OD600)范围内时,随着初始细胞密度的增大,菌株NNC-3降解苯酚的平均速率迅速增... 以Corynebacterium sp.NNC-3为受试菌株,就初始细胞密度对苯酚降解反应的影响进行了研究。结果表明,初始苯酚浓度为100~600 mg/L,初始细胞密度在0.05~0.30(OD600)范围内时,随着初始细胞密度的增大,菌株NNC-3降解苯酚的平均速率迅速增加;进一步增大初始细胞密度,菌株NNC-3降解苯酚的速率不再随初始细胞密度的增大而升高,菌株NNC-3降解苯酚的能力趋于饱和状态;初始苯酚浓度为600~800 mg/L时,菌株NNC-3降解苯酚的能力趋于饱和状态,且苯酚的降解速率呈现逐渐下降趋势,表明此时底物浓度为菌株NNC-3降解苯酚的限制性因素。研究成果对高效处理受酚污染环境具有一定的参考价值。 展开更多
关键词 细胞密度 苯酚 生物降解 棒杆菌NNC-3
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谷氨酸棒杆菌(Corynebacterium glutamicum)海藻糖合成酶的定点突变及其酶学性质研究
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作者 朱绮霞 陈发忠 +4 位作者 罗兆飞 韦宇拓 杜丽琴 王青艳 黄日波 《广西科学》 CAS 2012年第2期169-173,共5页
以已知晶体结构的Pseudomonas mesoacidophila MX-45菌株海藻酮糖合成酶(MutB)的晶体结构为模板,在SWISS-MODEL模建立谷氨酸棒杆菌(Corynebacterium glutamicum)海藻糖合成酶的立体结构,并对初始结构作能量优化,通过氨基酸序列比对,选择... 以已知晶体结构的Pseudomonas mesoacidophila MX-45菌株海藻酮糖合成酶(MutB)的晶体结构为模板,在SWISS-MODEL模建立谷氨酸棒杆菌(Corynebacterium glutamicum)海藻糖合成酶的立体结构,并对初始结构作能量优化,通过氨基酸序列比对,选择TreS-glu保守区内的氨基酸R245、D247、E289、F244和保守区外的氨基酸A288进行定点突变,并对突变酶F244C、F244L、F244W、F244Y、A288G、R245X、E289X、D247N、D247E进行纯化和酶学性质研究,比较突变子对酶活性和热稳定性的影响。结果表明,R245、E289突变为其它的19个氨基酸后酶活力全部丧失,D247E和D247N也丧失酶活,F244C、F244L、F244W、F244Y和A288G的比活力分别降低到TreS-glu的38%、24%、62%、64%和35%,A288突变成T288后没有酶活。与TreS-glu相比,F244C、F244W、A288G的Km值基本不变,F244L、F244Y对底物麦芽糖的亲和力降低,F244Y的最适反应温度和TreS-glu相同,均为27℃,而F244C、F244L、F244W和A288G的最适温度提高到32℃。与TreS-glu相比,突变酶的最适反应pH值均有所下降,其中F244C、F244Y和A288G的为7.5,比TreS-glu的8.0均下降了约0.5个单位,而F244L和F244W的为6.5,比TreS-glu的8.0均下降了近1.5个单位。与TreS-glu相比,突变酶的热稳定性均有不同程度提高,其中F244Y、F244W和A288G的Tm值比TreS-glu的提高约1℃,F244L提高约2℃,F244C提高了近4℃。 展开更多
关键词 海藻糖合成酶 谷氨酸棒杆菌 定点突变 酶学性质
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Engineering Corynebacterium glutamicum for Geraniol Production 被引量:1
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作者 Man Li Shuo Xu Wenyu Lu 《Transactions of Tianjin University》 2021年第5期377-384,共8页
Geraniol is a monoterpenoid alcohol with various applications in food,cosmetics,and healthcare.Corynebacterium glutamicum is a potential platform for terpenoids production because it harbors the methylerythritol phosp... Geraniol is a monoterpenoid alcohol with various applications in food,cosmetics,and healthcare.Corynebacterium glutamicum is a potential platform for terpenoids production because it harbors the methylerythritol phosphate pathway.To engineer C.glutamicum to produce geraniol,two different truncated geraniol synthases (GESs) were respectively expressed,and strain LX02 expressing the truncated GESs from Valeriana officinalis (t Vo GES) produced 0.3 mg/L of geraniol.Then,three geranyl diphosphate synthases (GPPSs) were combinatorially co-expressed with t Vo GES to improve geraniol production.The amounts of produced geraniol were all higher than that produced by strain LX02.Strain LX03 co-expressing ERG20 F96W–N127W (ERG20 WW) and t Vo GES produced the highest amount,5.4 mg/L.Subsequently,the co-overexpression of1-deoxy-D-xylulose-5-phosphate synthase (dxs) and isopentenyl diphosphate isomerase (idi) further increased the production to 12.2 mg/L in strain LX03.Lastly,the production of geraniol was increased to 15.2 mg/L via fermentation optimization.To our knowledge,this is the first report on the engineering of C.glutamicum to produce geraniol and thus can serve as a reference for other monoterpenoid production studies. 展开更多
关键词 corynebacterium glutamicum GERANIOL Geraniol synthase OVEREXPRESSION Metabolic engineering
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Effect of Lanthanon on Lysine Fermentation in Corynebacterium pekinense
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作者 Ying ZHANG 《Agricultural Biotechnology》 CAS 2012年第2期40-42,共3页
[ Objective] This study aimed to investigate the effects of rare earth element lanthanon on the growth and product synthesis of Corynebacterium pekinense. [ Method ] A full factorial experiment was designed to compare... [ Objective] This study aimed to investigate the effects of rare earth element lanthanon on the growth and product synthesis of Corynebacterium pekinense. [ Method ] A full factorial experiment was designed to compare the lysine fermentation of C. pek/nense in medium containing different concentrations of lanthanon. [ Result ] Low doses of lanthanum can promote the growth of C. pekinense and accumulation of metabolite lysine; optimization for distinct components of medium were obtained as 78 mg/L rare earth, 36 g/L glucose, 4.5 g/L beef extract, 20 g/L peptone by Matlab software. [Conclusion] The study initially deter- mined the optimum La+ additive amount in C. pekinense culture medium and the optimum medium compositions for C. pekinense to produce lysine. 展开更多
关键词 Rare earth elements corynebacterium pekinense Lysine fermentation
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Sub-acute endocarditis by Corynebacterium straitum : An often ignored pathogen
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作者 Vratika Agarwal Valay Parikh +3 位作者 Mayur Lakhani Chitradeep De Apurva Motivala Neville Mobarakai 《World Journal of Clinical Infectious Diseases》 2014年第1期1-4,共4页
With the emergence of novel etiologic organisms, panresistance, and invasive medical care infective endocarditis continues to be evasive, requiring newer approaches and modified treatment guidelines. Presented here is... With the emergence of novel etiologic organisms, panresistance, and invasive medical care infective endocarditis continues to be evasive, requiring newer approaches and modified treatment guidelines. Presented here is the case of a 75-year-old male with history of systolic heart failure with an automatic internal cardioverter defibrillator(AICD) implantation and a prosthetic mitral valve who presented with generalized malaise and progressive shortness of breath for 6 d. He was found to have positive blood cultures for gram positive rod shaped bacteria identified as Corynebacterium straitum, but was not considered as the etiological pathogen initially as it a usual skin contaminant. Later this bacterium was found to be the causative agent for the patient's endocarditis. This case highlights the importance of identifying the role of this uncommon commensal in invasive disease. With the use of effective antibiotic regimen and awareness of these new pathogens in invasive disease, mortality and morbidity can be prevented with initiation of early appropriate therapy. 展开更多
关键词 corynebacterium straitum ENDOCARDITIS Commensals CATHETER associated INFECTIONS IATROGENIC
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Aortic Graft Complicated by a Corynebacterium Striatum Infection Due to Previous Type IV Thoraco Abdominal Aortic Aneurysm Repair
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作者 Roberto Gabrielli Maria Sofia Rosati +2 位作者 Giovanni Caselli Alessandro Carra Andrea Siani 《World Journal of Cardiovascular Surgery》 2011年第2期5-10,共6页
We report successful management of aortic graft infection without graft explanation or extra-anatomic bypass. A 61 year-old male who had undergone surgical repair of a type IV thoraco-abdominal aortic aneurysm present... We report successful management of aortic graft infection without graft explanation or extra-anatomic bypass. A 61 year-old male who had undergone surgical repair of a type IV thoraco-abdominal aortic aneurysm presented with left flank pain and raised inflammatory markers following graft implantation. CT scanning revealed a left psoas fluid collection. Graft infection was diagnosed. A radio-guided drainage and conservative management, with irrigation drain placement was undertaken with preservation of the aortic graft. There was no evidence of recurrent infection after follow-up at 34 months. Aortic endograft infection may be managed by surgical or radio-guided drainage, antibiotic irrigation of the graft and systemic antibiotic therapy without graft removal. 展开更多
关键词 AORTIC GRAFT INFECTION corynebacterium STRIATUM Antibiotic IRRIGATION
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Human Seroreactivity to Secreted Molecules of Corynebacterium pseudotuberculosis
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作者 Ramon Mendes dos Santos Silvânia Maria Andrade Cerqueira +16 位作者 Caio Lopez Borge Andrade Gabriel Saldanha Müller Vitória Costa de Menezes Santos Hévlyn Ribeiro de Araújo Samanta Queiroz Rogério Reis Conceição Luiz Gustavo Freitas Oliveira Marcos Borges Ribeiro Silvana Marchioro Lilia Ferreira de Moura-Costa Fúlvia Soares Campos de Sousa Maria da Conceição Aquino de Sá José Tadeu Raynal Rocha Filho Soraya Castro Trindade Eduardo Martins Netto Roberto Meyer Songelí Menezes Freire 《Advances in Microbiology》 2022年第3期150-158,共9页
Corynebacterium pseudotuberculosis is an infectious agent that occurs in small ruminants causing caseous lymphadenitis, and more rarely in humans causing lymphadenitis and pneumonia. The breeding small ruminants have ... Corynebacterium pseudotuberculosis is an infectious agent that occurs in small ruminants causing caseous lymphadenitis, and more rarely in humans causing lymphadenitis and pneumonia. The breeding small ruminants have great economic importance in Brazil. Rural farm workers and veterinary students who acquired this disease suffered from weakening symptoms for weeks, and the identification of the etiological agent was time-consuming and complex. Due to the low prevalence of case records, there is probably no available commercial diagnostic kit for C. pseudotuberculosis infection in humans. This study aimed to describe human seroreactivity to secreted antigens from C. pseudotuberculosis. Reactivity of serum from farm workers (n = 14), individuals who work with the bacillus at laboratory (n = 8) or individuals without contact (n = 25) was tested with secreted proteins from PAT10 strain of C. pseudotuberculosis by Western blotting. Samples of all (100%) farm workers showed reactivity to 31 kDa, 71 kDa and 164 kDa proteins, while laboratory workers showed 87.5%, 62.5 % and 37.5%, and no-contact 20%, 0% and 16%, respectively. All sera recognized the 275 kDa protein. Our data suggest that C. pseudotuberculosis secreted proteins are antigenic in humans and the recognition profiles allowed the identification of individuals with and without prior contact with this bacillus. This is the first paper which describes human reactivity to C. pseudotuberculosis in serum samples of workers in Brazil. 展开更多
关键词 Human Immunoreactivity ANTIGENICITY corynebacterium pseudotuberculosis Secreted Proteins Western Blot
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利用CRISPRi技术构建乙醛酸生物合成Corynebacterium glutamicum工程菌 被引量:1
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作者 梁咏思 沈凯佳 +2 位作者 范许云 韩武洋 李天明 《食品科学》 EI CAS CSCD 北大核心 2021年第2期170-176,共7页
以谷氨酸棒状杆菌(Corynebacterium glutamicum ATCC 13032)为出发菌株,敲除其支流代谢关键酶乳酸脱氢酶合成基因lldh,建立规律间隔成簇短回文重复序列干扰(clustered regularly interspaced short palindromic repeats interference,CR... 以谷氨酸棒状杆菌(Corynebacterium glutamicum ATCC 13032)为出发菌株,敲除其支流代谢关键酶乳酸脱氢酶合成基因lldh,建立规律间隔成簇短回文重复序列干扰(clustered regularly interspaced short palindromic repeats interference,CRISPRi)调控体系,并利用该体系下调支流代谢中的关键酶异柠檬酸脱氢酶合成基因icd和苹果酸合成酶合成基因ms的表达强度,同时过表达异柠檬酸裂合酶合成基因icl,强化乙醛酸合成的通路。通过48 h连续监测工程菌和野生菌生长状况,并检测发酵终产物。结果显示:工程菌生长几乎不受影响,发酵液中乙醛酸质量浓度达到5 mg/mL,实现了乙醛酸的积累,为谷氨酸棒状杆菌工业生产乙醛研究提供一定的参考。 展开更多
关键词 谷氨酸棒状杆菌 乙醛酸 规律间隔成簇短回文重复序列干扰 工程菌
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北京棒杆菌Corynebacterium pekinense天冬氨酸激酶P184M酶动力学分析及酶学性质表征
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作者 王锐楠 张芷睿 +2 位作者 方丽 闵伟红 刘春雷 《食品安全质量检测学报》 CAS 2017年第5期1750-1756,共7页
目的研究北京棒杆菌天冬氨酸激酶(aspartate kinase,AK)184位点脯氨酸(proline,P)突变为蛋氨酸(DL-methionine,M)对酶动力学及酶学性质的影响。方法对北京棒杆菌AK家族同源序列比对,选择出高度保守且远离底物结合位点的Pro184位点,对其... 目的研究北京棒杆菌天冬氨酸激酶(aspartate kinase,AK)184位点脯氨酸(proline,P)突变为蛋氨酸(DL-methionine,M)对酶动力学及酶学性质的影响。方法对北京棒杆菌AK家族同源序列比对,选择出高度保守且远离底物结合位点的Pro184位点,对其进行饱和定点突变,成功构建突变体P184M。结果动力学和酶学性质研究表明:P184M AK Vmax比WT(wide type)提高了5.06倍;n值为0.19,突变体由WT的正协同性变为负协同性,同时Km(mol/L)值增大;突变体AK最适pH为6.5,低于野生型最适pH 7.0;最适反应温度28℃,低于WT的30℃,适宜温度区间变窄;半衰期由WT的2.1 h降为1.2 h;金属离子、有机溶剂和抑制剂Thr+Met、Lys+Met、Thr+Lys+Met对突变株AK均表现出激活作用。结论突变体天冬氨酸激酶P184M的酶动力学性质和酶学性质改变显著,为构建高产蛋氨酸菌株提供了一定的参考依据。 展开更多
关键词 北京棒杆菌 天冬氨酸激酶 突变株 酶学性质
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Functional food additives/ingredients production by engineered Corynebacterium glutamicum
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作者 Katarina Cankar Nadja A.Henke Volker F.Wendisch 《Systems Microbiology and Biomanufacturing》 2023年第1期110-121,共12页
Corynebacterium glutamicum is a microbial production host established in the industry 60 years ago.It is mainly used for production of feed and food amino acids.As C.glutamicum strain development has been cutting edge... Corynebacterium glutamicum is a microbial production host established in the industry 60 years ago.It is mainly used for production of feed and food amino acids.As C.glutamicum strain development has been cutting edge since its discovery,it has been engineered for production of a plethora of valuable products.This review will focus on recent developments of C.glutamicum strain engineering for biotransformation and fermentation processes towards flavor and fragrance molecules as well as pigments and sweeteners. 展开更多
关键词 corynebacterium glutamicum FLAVORS FRAGRANCES PIGMENTS SWEETENERS
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PVA固定化Corynebacterium sp.JY03降解苯酚的特性研究 被引量:8
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作者 姜立春 阮期平 王晓丽 《环境工程》 CAS CSCD 北大核心 2014年第2期36-40,共5页
以PVA(聚乙烯醇)作为载体将降酚菌株Corynebacteriumsp.JY03进行固定化包埋处理,正交试验确定该菌株固定化细胞制备的条件,然后对固定化细胞的降酚性能进行研究。试验确定最佳固定条件为:PVA质量分数为6%,菌液量/PVA水溶液体... 以PVA(聚乙烯醇)作为载体将降酚菌株Corynebacteriumsp.JY03进行固定化包埋处理,正交试验确定该菌株固定化细胞制备的条件,然后对固定化细胞的降酚性能进行研究。试验确定最佳固定条件为:PVA质量分数为6%,菌液量/PVA水溶液体积比为6/30,氯化钙含量为2.0%,钙化交联时间为8h;固定化细胞降解苯酚的最适温度为30~35℃,最适pH值为6.5~7.5,在初始苯酚浓度为700mg/L,装液量50mL,培养42h后,苯酚降解率达99.1%。固定化细胞重复利用6次苯酚降解率仍高于85.2%,其性能优于游离细胞,这将为该菌株进一步应用于含酚废水的生物处理提供实践条件。 展开更多
关键词 corynebacterium SP JY03 苯酚 PVA 固定化细胞 降酚性能
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Etiological Agent, Pathogenesis, Diagnosis, Treatment, Measures for Prevention and Control of Caseous Lymphadenitis Disease in the Small Ruminants with Special Reference to Sheep
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作者 Eréndira de la Fuente Mancera Abel Ciprian Carrasco Susana Mendoza Elvira 《Journal of Biosciences and Medicines》 2024年第5期154-170,共17页
Caseous lymphadenitis (CL) is a chronic infectious disease caused by Corynebacterium pseudotuberculosis (C. pseudotuberculosis) that is transmitted mainly by contact with contaminated exudate wounds. Caseous lymphaden... Caseous lymphadenitis (CL) is a chronic infectious disease caused by Corynebacterium pseudotuberculosis (C. pseudotuberculosis) that is transmitted mainly by contact with contaminated exudate wounds. Caseous lymphadenitis affects different species of animals and is considered an occupational zoonotic disease. CL is responsible for important economic losses in the small ruminant industry, which include decreased production, damaged quality of milk and wool, reproductive disorders, total or partial confiscation of carcasses, and depreciation of the skin. Caseous lymphadenitis disease can present in two clinical pictures: a skin or superficial picture and a visceral or deep picture. The presumptive diagnosis of CL in the skin is based on the exploration of superficial lymph nodes. These have little value in diagnosing visceral CL, where the main sign is emaciation. The prevention and control of CL involve identifying the skin condition and debriding the abscesses. In addition to the treatment of superficial lesions, the animal facilities must be repaired and adapted. To avoid exposure of negative herds to CL, new animals must be subjected to observation to identify any of the clinical manifestations. Another form of control is vaccination, though only some countries have commercial vaccines. General information on the etiological agent and its characteristics can be used to improve the understanding of this disease. 展开更多
关键词 ABSCESS CMNR corynebacterium pseudotuberculosis GOAT CL Lymph Nodes PSEUDOTUBERCULOSIS SHEEP
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