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1.
从农药厂废水中分离到6株能以除草剂阿特拉津为唯一氮源生长的细菌,即假单胞菌(Pseu-domonas spp.)AD1、AD2和 AD6,土壤杆菌(Agrobacterium sp.)AD4,黄单胞菌(Xanthomonas sp.)ADS,欧文氏菌(Erwinia sp.)AD7。AD1菌株能使无机盐培养基中的 0.3g/L阿特拉津在72h内降解99,9%。当以AD1、AD2、AD4、AD5、AD6和AD7菌株的总DNA为模板进行PCR扩增时,除A  相似文献   

2.
高产稳产聚羟基烷酸的重组大肠杆菌的构建   总被引:7,自引:0,他引:7  
重组大肠杆菌Escherichia coliHMS174(pTZ18UPHB) 含有携带聚羟基烷酸(PHA) 合成基因( phaCAB)** 的质粒pTZ18UPHB,是很有潜力的PHA 生产菌,但存在着质粒不稳定和不能合成3羟基丁酸(3HB) 与3羟基戊酸(3HV) 共聚物[P(3HBco3HV)] 的缺陷。将RK2 质粒上的par DE 基因引入pTZ18UPHB 构成质粒pJMC2 ,该质粒可以在宿主E.ColiHMS174 中稳定遗传。将培养基中的磷酸盐浓度降至18 m mol/L,发现E.Coli HMS174(pJMC2) 能够以丙酸为前体合成P(3HBco3HV) ,其中3HV 在共聚物中的含量为5 % ~8 % 。在5L自动发酵罐中分批补料培养E.Coli HMS174(pJMC2) ,培养基初始磷酸盐浓度为15 m mol/L,30 h 后每升培养液中干菌体可达42-5 g,P(3HBco3HV) 占干重的70 % ,其中3HV 在共聚物中的含量为4-9 % 。  相似文献   

3.
对表达人骨形成蛋白2A(BMP2A)的重组大肠杆菌YK537/pDHB2m在500ml摇瓶中进行了培养条件的摸索实验,继后用5L自控发酵罐进行分批培养和分批补料培养,以获取rhBMP2A。两种培养方式结果比较表明,在培养过程中保持30%~40%左右的溶解氧和限制性流加葡萄糖可以使BMP2A的含量达到278g/L,最终菌体密度为OD60053(相当于干菌212g/L),重组蛋白的表达量占菌体总蛋白的25%。该培养技术的关键是:(1)在培养过程中保持适当的溶解氧;(2)限制性流加葡萄糖;(3)42℃起始诱导的时间控制在对数生长中期,持续表达时间为4h;(4)细菌持续生长的比生长速率控制在03h1左右。  相似文献   

4.
在数值分类、SDSPAGE 全细胞蛋白分析、DNADNA 杂交、16SrDNAPCRRFLP 的基础上,测定了两个分离自干旱地区苜蓿、草木樨根瘤菌新群1 、2 的中心株XJ96060 、XJ96408 的16SrDNA 全序列,并进一步将中心株和31 株已知菌、3 株分自黄土高原的根瘤菌进行了系统发育学分析。结果表明,供试菌株在系统发育树中基本分成Sinorhizobium 、Mesorhizobium 、AgrobacteriumRhizobium 、Rhizobiu m 、Bradyrhizobium 、Azorhizobium 六个分枝。群1 ,2 落入Sinorhizobiu m 分枝。  相似文献   

5.
分离的酒色着色菌(Chromatiumvinosum)内膜系统在光照和O乙酰丝氨酸(OAS)存在的条件下,能以359纳摩尔/毫克细菌叶绿素·小时(nmol·mgBchl-1·h-1)的速度催化SeO2-3合成硒半胱氨酸。用超声波处理的内膜系统,催化速度仅为处理前的11%,加入谷胱甘肽(GSH)和还原型辅型Ⅱ(NADPH)后,其速度增加至处理前的883%,该反应对光具有依赖性,进一步实验表明,纯化的谷胱甘肽还原酶,在有半胱氨合酶、OAS和NADPH共存时,能催化SeO2-3转化为硒半胱氨酸,表明SeO2-3在内膜系统中能被光偶联的谷胱甘肽还原酶还原为Se2-,然后经半胱氨酸合酶的催化作用转化为硒半胱氨酸  相似文献   

6.
从不同来源的细菌菌株筛选获得一株吸附还原Au3+较强的菌株D01,经鉴定为巨大芽孢杆菌(Bacilusmegatherium)D01。菌株D01在Au3+浓度600mg/L下仍能较好生长。从电化学反应表明,该菌具有较强的还原力,它能将金催化剂的前驱体Au3+/αFe2O3还原成具有催化CO+O2→CO2的高分散度的Au0/αFe2O3催化剂  相似文献   

7.
建立了脱脂奶溶蛋白琼脂扩散试验新方法,用于检测嗜水气单胞菌(Aeromonas hydrophila Ah)J-1株两种培养基培养的去菌体上清中的胞外蛋白酶(ECP),同时用经典底物法进行了检测,两种方法所得结果相符。与底物法相比,脱脂奶溶蛋白琼脂扩散试验操作简便、敏感性高、重复性好。  相似文献   

8.
中华鳖白底板病和红底板病细菌的分离鉴定及致病性   总被引:4,自引:0,他引:4  
从患白底板病和红底板病的病鳖中分离出7株细菌,应用一般细菌分离鉴定的方法和美国生物-梅里埃(Bio Merieux USA)公司VITEK全自动微生物鉴定系统进行鉴定,结果显示有3株嗜水气单胞菌(Aeromonas hydrophila)、1株温和气单胞菌(Aeromonas sobria<.I>)、1株金黄色葡萄球菌(Staphylococcusaureus)、1株肺炎克雷伯氏菌(Klebsiella pneumoniae)和 1株美人鱼弧菌(Vib  相似文献   

9.
枯草芽孢杆菌B034拮抗蛋白的分离纯化及特性分析   总被引:29,自引:2,他引:29  
枯草芽孢杆菌(Bacilussubtilis)B034分离自水稻叶面,对水稻白叶枯病菌具有较强的拮抗能力。除去菌体培养液以70%饱和度硫酸铵沉淀所得的拮抗物粗提液对热稳定,对胰蛋白酶不敏感,对蛋白酶K、链霉蛋白酶E部分敏感,对氯仿部分敏感,其作用的活性pH范围低至4,高至12以上,比较耐碱性。粗提液经PhenylSepharoseCL4B柱层析、DEAESephacel柱层析和HPLC的Superdex75HR10/30柱层析,得到二个拮抗活性峰:P1和P2。P2经SDSPAGE和PAGEIEF电泳显示为单一蛋白带,分子量503kD,等电点625。自动Edman降解法从P2的N端测出残基序列为IleSerAsnProXIleAspVal  相似文献   

10.
产β1 ,4D甘露聚糖酶的诺卡氏菌形放线菌( Nocardioform actinomycetes) 菌株NA3540 ,发酵培养72h ,发酵液离心去菌体,上清经硫酸铵沉淀,95 % 乙醇沉淀,CMSephadex A50 柱层析、羟基磷灰石柱层析、DEAE纤维素离子交换及Sephadex G100 分子凝胶过滤柱等步骤,β甘露聚糖酶的比活提高了137 倍,获得凝胶电泳均一的蛋白样品。经SDSPAGE 和凝胶过滤法分别测定β甘露聚糖酶分子量为41kD和40kD,证明该酶为单聚体;用等电聚焦电泳测得其等电点为4-8 ;经氨基酸组成分析,发现蛋白中有较高含量的Gly、Asp、Ala 及Glu 残基。该酶的最适反应温度为75 ℃,在不超过60 ℃时酶活较稳定;酶反应的最适pH 为8-0 ,pH 稳定范围为6-5~11-0 。重金属离子Hg2+ 、Cu2+ 、Pb2+ 、Fe3+ 、Co2+ 、Zn2+ 能强烈抑制该酶活性,而Mn2+ 、Fe2+ 、Ag 对该酶有部分的抑制作用,低浓度的Na 、K 、Li 对该酶基本没有影响。  相似文献   

11.
The nematode Heterorhabditis bacteriophora transmits a monoculture of Photorhabdus luminescens bacteria to insect hosts, where it requires the bacteria for efficient insect pathogenicity and as a substrate for growth and reproduction. Siderophore production was implicated as being involved in the symbiosis because an ngrA mutant inadequate for supporting nematode growth and reproduction was also deficient in producing siderophore activity and ngrA is homologous to a siderophore biosynthetic gene, entD. The role of the siderophore in the symbiosis with the nematode was determined by isolating and characterizing a mini-Tn5-induced mutant, NS414, producing no detectable siderophore activity. This mutant, being defective for growth in iron-depleted medium, was normal in supporting nematode growth and reproduction, in transmission by the dauer juvenile nematode, and in insect pathogenicity. The mini-Tn5 transposon was inserted into phbH; whose protein product is a putative peptidyl carrier protein homologous to the nonribosomal peptide synthetase VibF of Vibrio cholerae. Other putative siderophore biosynthetic and transport genes flanking phbH were characterized. The catecholate siderophore was purified, its structure was determined to be 2-(2,3-dihydroxyphenyl)-5-methyl-4,5-dihydro-oxazole-4-carboxylic acid [4-(2,3-dihydroxybenzoylamino)-butyl]-amide, and it was given the generic name photobactin. Antibiotic activity was detected with purified photobactin, indicating that the siderophore may contribute to antibiosis of the insect cadaver. These results eliminate the lack of siderophore activity as the cause for the inadequacy of the ngrA mutant in supporting nematode growth and reproduction.  相似文献   

12.
The nematode Heterorhabditis bacteriophora transmits a monoculture of Photorhabdus luminescens bacteria to insect hosts, where it requires the bacteria for efficient insect pathogenicity and as a substrate for growth and reproduction. Siderophore production was implicated as being involved in the symbiosis because an ngrA mutant inadequate for supporting nematode growth and reproduction was also deficient in producing siderophore activity and ngrA is homologous to a siderophore biosynthetic gene, entD. The role of the siderophore in the symbiosis with the nematode was determined by isolating and characterizing a mini-Tn5-induced mutant, NS414, producing no detectable siderophore activity. This mutant, being defective for growth in iron-depleted medium, was normal in supporting nematode growth and reproduction, in transmission by the dauer juvenile nematode, and in insect pathogenicity. The mini-Tn5 transposon was inserted into phbH; whose protein product is a putative peptidyl carrier protein homologous to the nonribosomal peptide synthetase VibF of Vibrio cholerae. Other putative siderophore biosynthetic and transport genes flanking phbH were characterized. The catecholate siderophore was purified, its structure was determined to be 2-(2,3-dihydroxyphenyl)-5-methyl-4,5-dihydro-oxazole-4-carboxylic acid [4-(2,3-dihydroxybenzoylamino)-butyl]-amide, and it was given the generic name photobactin. Antibiotic activity was detected with purified photobactin, indicating that the siderophore may contribute to antibiosis of the insect cadaver. These results eliminate the lack of siderophore activity as the cause for the inadequacy of the ngrA mutant in supporting nematode growth and reproduction.  相似文献   

13.
CAS平板覆盖法检测氢氧化细菌铁载体   总被引:3,自引:1,他引:2  
【目的】用CAS平板覆盖法检测氢氧化细菌铁载体,解决通用CAS琼脂平板法中十六烷基三甲基溴化铵对真菌和某些细菌的生长抑制问题。【方法】将改良的CAS检测培养基覆盖在长满菌落的无铁培养基上,生长抑制问题因微生物未与十六烷基三甲基溴化铵直接接触而解决。【结果】3株氢氧化细菌SDW-5、SDW-9和AaP-13均能产生单菌落,加入CAS检测培养基1 h后,菌落周围产生明显的铁载体晕圈。【结论】本方法成功解决了生长抑制问题,可以作为检测微生物铁载体的通用方法。  相似文献   

14.
Iron acquisition mechanisms play an important role in the pathogenesis of many infectious microbes. In Bacillus anthracis, the siderophore petrobactin is required for both growth in iron‐depleted conditions and for full virulence of the bacterium. Here we demonstrate the roles of two putative petrobactin binding proteins FatB and FpuA (encoded by GBAA5330 and GBAA4766 respectively) in B. anthracis iron acquisition and pathogenesis. Markerless deletion mutants were created using allelic exchange. The ΔfatB strain was capable of wild‐type levels of growth in iron‐depleted conditions, indicating that FatB does not play an essential role in petrobactin uptake. In contrast, ΔfpuA bacteria exhibited a significant decrease in growth under low‐iron conditions when compared with wild‐type bacteria. This mutant could not be rescued by the addition of exogenous purified petrobactin. Further examination of this strain demonstrated increased levels of petrobactin accumulation in the culture supernatants, suggesting no defect in siderophore synthesis or export but, instead, an inability of ΔfpuA to import this siderophore. ΔfpuA spores were also significantly attenuated in a murine model of inhalational anthrax. These results provide the first genetic evidence demonstrating the role of FpuA in petrobactin uptake.  相似文献   

15.
The growth of marine bacteria under iron-limited conditions was investigated. Neither siderophore production nor bacterial growth was detected for Pelagiobacter sp. strain V0110 when Fe(III) was present in the culture medium at a concentration of <1.0 microM. However, the growth of V0110 was strongly stimulated by the presence of trace amounts of exogenous siderophore from an alpha proteobacterium, V0902, and 1 nM N-acyl-octanoylhomoserine lactone (C(8)-HSL), which is known as a quorum-sensing chemical signal. Even though the iron-binding functionality of a hydroxamate siderophore was undetected in the supernatant of V0902, a hydroxamate siderophore was detected in the supernatant of V0110 under the above conditions. These results indicated that hydroxamate siderophore biosynthesis by V0110 began in response to the exogenous siderophore from V0902 when in the presence of C(8)-HSL; however, C(8)-HSL production by V0110 and V0902 was not detected. Direct interaction between V0902 and V0110 through siderophore from V0902 was observed in the dialyzing culture. Similar stimulated growth by exogenous siderophore and HSL was also observed in other non-siderophore-producing bacteria isolated from marine sponges and seawater. The requirement of an exogenous siderophore and an HSL for heterologous siderophore production indicated the possibility that cell-cell communication between different species was occurring.  相似文献   

16.
嗜水气单胞菌生物被膜对其耐药性的影响   总被引:9,自引:0,他引:9  
建立了嗜水气单胞菌(Aeromonas hydrophila, Ah)的生物被膜(Bacterial biofilm, BF) 体外形成模型, 并对11种抗菌药物对BF细菌和浮游(Freecell, FC)细菌的清除作用进行了研究。将Ah J1株在放有硅胶膜的TSB中培养7d,用银染法鉴定,发现可形成良好的BF。FC细菌对青霉素具有耐药性, 最低杀菌浓度(MBC)为256μg/mL;对蒽诺沙星和氟哌酸最敏感,MBC分别为003μg/mL和0.25μg/mL。氟苯尼考对BF细菌的清除能力最强,作用于BF细菌和FC细菌的MBC之比为2∶1;卡那霉素、青霉素、新霉素的MBC比值在32∶1以上。扫描电镜观察蒽诺沙星作用于FC及BF细菌前后的形态变化,并测定其杀菌曲线。发现4×MBC时可完全清除FC细菌,但不能完全清除BF细菌;在32×MBC时,4h内可完全清除FC细菌,而24h内完全清除BF细菌。结果表明形成BF的Ah对抗菌药物可形成强耐受性,其潜在影响应引起足够重视。  相似文献   

17.
烟草根际铁载体产生菌G-229-21T的筛选、鉴定及拮抗机理   总被引:5,自引:0,他引:5  
[目的]从烟草根际筛选烟草疫霉[Phytophthora parasitica var.nicotianae(Breda de Hann)Tucker]拮抗菌,探索其拮抗机理.[方法]限铁(2.0 μmol/L FeCl3)蔗糖-天冬酰胺平板对峙法筛选烟草疫霉拮抗菌;刃天青(CAS)法检测其铁载体的产生及其对铁离子的亲和能力.结合形态、生理生化、16s rRNA序列同源性和系统发育分析及种特异性分子法对其进行鉴定.XAD-2吸附层析法提取其铁载体,分光光度法检测其铁载体类型.不同铁离子浓度下,比较其铁载体对烟草疫霉的抑制作用.[结果]我们筛选到一株限铁条件下烟草疫霉拮抗菌G-229-21T,该菌产生高亲和力铁载体,被初步鉴定为Pseudomonas mediterranea.该菌产生的羧酸型铁载体,在低铁条件下(0.16μmol/L~10μmol/L,FeCl3)对烟草疫霉的抑制率达92.3%以上,而在富铁条件下(100 μmol/L FeCl3)抑制率仅为2.0%.[结论]首次报道P. mediterranea G-229-21T产生高亲和力羧酸型铁载体,该铁载体在低铁条件下对烟草疫霉有显著的抑制作用.  相似文献   

18.
Since N-acyl homoserine lactones (AHLs) are key mediators of cell density-dependent regulation of traits involved in virulence and epiphytic fitness in gram-negative bacteria such as Pseudomonas syringae, a variety of plant species were examined to determine their production of leaf surface compounds that could interact with these signaling systems. Leaf washings of 17 of 52 plant species tested stimulated or inhibited AHL-dependent traits in at least one of the bacterial reporter strains used. The active compounds from most plants could be distinguished from known AHLs due to different patterns of mobility during C8 and C18 reverse-phase thin-layer chromatography (TLC) and normal-phase TLC compared to the patterns for authentic bacterial AHLs. All plant extracts were also tested to determine their abilities to sequester iron and trigger bacterial siderophore synthesis on a medium containing abundant iron. Leaf washings from 16 of the 52 plant species, as well as tannic acid solutions, stimulated pyoverdine synthesis in P. syringae in a high-iron medium. These preparations also inhibited the growth of a P. syringae mutant unable to produce pyoverdine siderophores but not the growth of the wild-type bacterium. The stimulation of siderophore production and the growth inhibition by plant extracts and purified tannins were both reversed by addition of ferric chloride to culture media, indicating that iron was made unavailable by the compounds released onto the leaf surface.  相似文献   

19.
The growth of marine bacteria under iron-limited conditions was investigated. Neither siderophore production nor bacterial growth was detected for Pelagiobacter sp. strain V0110 when Fe(III) was present in the culture medium at a concentration of <1.0 μM. However, the growth of V0110 was strongly stimulated by the presence of trace amounts of exogenous siderophore from an alpha proteobacterium, V0902, and 1 nM N-acyl-octanoylhomoserine lactone (C8-HSL), which is known as a quorum-sensing chemical signal. Even though the iron-binding functionality of a hydroxamate siderophore was undetected in the supernatant of V0902, a hydroxamate siderophore was detected in the supernatant of V0110 under the above conditions. These results indicated that hydroxamate siderophore biosynthesis by V0110 began in response to the exogenous siderophore from V0902 when in the presence of C8-HSL; however, C8-HSL production by V0110 and V0902 was not detected. Direct interaction between V0902 and V0110 through siderophore from V0902 was observed in the dialyzing culture. Similar stimulated growth by exogenous siderophore and HSL was also observed in other non-siderophore-producing bacteria isolated from marine sponges and seawater. The requirement of an exogenous siderophore and an HSL for heterologous siderophore production indicated the possibility that cell-cell communication between different species was occurring.  相似文献   

20.
Under conditions of iron limitation Pseudomonas fluorescens ATCC 17400 produces two siderophores, pyoverdine, and a second siderophore quinolobactin, which itself results from the hydrolysis of the unstable molecule 8-hydroxy-4-methoxy-2-quinoline thiocarboxylic acid (thioquinolobactin). Pseudomonas fluorescens ATCC 17400 also displays a strong in vitro antagonism against the Oomycete Pythium, which is repressed by iron, suggesting the involvement of a siderophore(s). While a pyoverdine-negative mutant retains most of its antagonism, a thioquinolobactin-negative mutant only slowed-down Pythium growth, and a double pyoverdine-, thioquinolobactin-negative mutant, which does not produce any siderophore, totally lost its antagonism against Pythium. The siderophore thioquinolobactin could be purified and identified from spent medium and showed anti-Pythium activity, but it was quickly hydrolysed to quinolobactin, which we showed has no antimicrobial activity. Analysis of antagonism-affected transposon mutants revealed that genes involved in haem biosynthesis and sulfur assimilation are important for the production of thioquinolobactin and the expression of antagonism.  相似文献   

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