绿色荧光蛋白标记的大肠杆菌海水中稳定性研究

用绿色荧光蛋白基因( GFP) 标记大肠杆菌K12(K12GFP),检测其在海水中的存活及稳定性.研究结果表明,K12与K12GFP微生物学特性差异不显著,而且两者在高压灭菌海水中的存活曲线也基本一致,说明K12GFP可为进一步研究K12在自然海水中的生态变化提供可靠依据.在高压灭菌海水中,K12GFP存活28 d之久,该菌密度随时间缓慢下降;在过滤海水中,K12GFP存活7 ,该菌密度从107CFU/mL降到102CFU/mL.表明海水的营养成分会对其存活产生较大影响.     

Analysis of antibiotic resistance of Escherichia coli isolated from the Yitong River in North-east China

The concentrations of four types of antibiotics in the Yitong River were detected. The concentration of total coliforms in summer was higher than that in spring. There was a seasonal difference in antibiotic resistance of E. coli. The E. coli in the Yitong River was found to have multiple antibiotic resistance. The Yitong River is one of the largest secondary tributaries of the Songhua River. The area where the Yitong River flows is densely populated and contains the livestock and poultry breeding areas of north-east China. These areas introduce a high risk of antibiotic contamination. In this study, the concentrations of four types of typical antibiotics including quinolones, tetracyclines, sulfonamides, and trimethoprim were determined by solid phase extraction-high performance liquid chromatography. The antibiotic resistance of Escherichia coli caused by antibiotic pollution was investigated. The concentration of total coliforms in the Yitong River was detected by the plate counting method. The antibiotic resistance of E. coli to quinolones, tetracyclines, sulfonamides, and trimethoprim was analyzed by the Kirby-Bauer method. The results showed that the concentration of total coliforms in the summer was higher than that in the spring. There was a seasonal difference in the resistance rate of E. coli to antibiotics except trimethoprim. The antibiotic resistance to fluoroquinolones was relatively low. The resistance rate to tetracyclines was higher during the summer. Moreover, resistance to several antibiotics was observed in all sections. This study provides basic data for research on pollution characteristics and prevention of antibiotic exposure in rivers.     

大肠杆菌植酸酶在毕赤酵母中的表达与纯化

为探讨植酸酶的结构与功能,研究了来源于大肠杆菌的植酸酶基因在酵母中的表达和纯化条件.将含有大肠杆菌植酸酶基因的毕赤酵母工程菌在不同甲醇浓度和不同诱导时间下培养,检测植酸酶的表达情况.结果表明:诱导培养基中一次性添加2.5%的甲醇,诱导96 h后,蛋白浓度达到1.36 mg mL-1,酶活力达到6 530 U mL-1;将在毕赤酵母中表达的植酸酶粗酶液经过硫酸铵盐析、Resource S柱和Superdex-75三步纯化,得到单峰纯的蛋白,比活力为137 280 Umg-1.用圆二色谱分析纯化后的蛋白在pH 5.0和8.0环境中的结构变化,结果显示pH 8.0环境使大肠杆菌植酸酶发生了变性.图3参16     

Deposition of copper nanoparticles on multiwalled carbon nanotubes modified with poly (acrylic acid) and their antimicrobial application in water treatment

A novel hybrid material, Cu-PAA/MWCNTs (copper nanoparticles deposited multiwalled carbon nanotubes with poly (acrylic acid) as dispersant, was prepared and expected to obtain a more effective and well-dispersed disinfection material for water treatment. X-ray energy dispersive spectroscopy (EDS), transmission electron microscopy (TEM), the X-ray fluorescence (XRF), X-ray photoelectron spectra (XPS), Fourier transform infrared spectra (FT-IR), Raman spectroscopy, and thermal gravimetric analyzer (TGA) were used to characterize the Cu-PAA/MWCNTs. Escherichia coli (E. coil) was employed as the target bacteria. The cell viability determination and fluorescence imaging results demonstrated that Cu-PAA/MWCNTs possessed strong antimicrobial ability on E. coil. The deposited Cu was suggested to play an important role in the antimicrobial action of Cu-PAA/MWCNTs.     

大肠杆菌与水体中U(Ⅵ)的作用行为和产物研究

通过批次吸附实验及介观和谱学等表征方法,研究了大肠杆菌(E.coli)粉末对水体中U(Ⅵ)的富集行为和吸附模型,并对其作用产物进行了详细分析.结果表明:大肠杆菌对初始浓度为50mg/L U(Ⅵ)溶液(pH＝5)的吸附容量可达到276.89mg/g.Langmuir等温模型和准二级动力学方程能较好的描述其吸附过程. FTIR、SEM-EDS、XRD分析结果表明:在与水体中U(VI)作用后,大肠杆菌表面检测出UO₂²⁺的红外特征峰(876.16cm^-1)和U的能谱吸收峰(结合能=2.4~4.4keV).UO₂²⁺主要与菌体表面的烷基、氨基、羧基、分子间氢键发生作用,重点与PO₂-、P(OH)₂、PO₄^3-以及PO₃^-等含P基团进行络合配位,最终产物以CaU(PO₄)₂、Ca(UO₂)₂(PO₄)₂·xH₂O、NaUO₂(PO₃)₃等铀的磷酸盐形式存在.     

Synthesis and antibacterial activity of a Fe3O4–AgCl nanocomposite against Escherichia coli

In this investigation, Fe₃O₄ magnetic nanoparticles (MNPs) were prepared by the alkalinization of an aqueous medium containing ferrous sulfate and ferric chloride. In the next step, a Fe₃O₄–AgCl magnetic nanocomposite was fabricated by the drop-by-drop addition of silver nitrate solution into a NaCl solution containing Fe₃O₄ MNPs. All prepared nanoparticles were characterized by transition electron microscopy (TEM), X-ray diffraction (XRD), and energy-dispersive X-ray spectroscopy (EDS). Both particle types varied in size from 2.5 to 20?nm, with an average size of 7.5?nm for Fe₃O₄ MNPs and 12.5?nm for Fe₃O₄–AgCl nanocomposites. The antibacterial effect of the Fe₃O₄ MNPs and fabricated Fe₃O₄–AgCl nanocomposites against Escherichia coli (ATCC 35218) were investigated by conventional serial agar dilution method using the Müller–Hinton Agar medium. The minimum inhibitory concentration was 4?mg?mL^?1 for Fe₃O₄ MNPs and 2?mg?mL^?1 for the Fe₃O₄–AgCl magnetic nanocomposites. Time-kill course assays showed that the Fe₃O₄–AgCl magnetic nanocomposites successfully killed all inoculated bacterial cells during an exposure time of 60?min. The antibacterial activity of recycled Fe₃O₄–AgCl magnetic nanocomposites over four 60?min cycles of antibacterial treatment was further tested against E. coli by the colony-forming unit (CFU) method. The antibacterial efficiency of the nanocomposites was constant over two cycles of antibacterial testing.     

Microbiological evaluation of 4-(6-chloro-7H-purin-8-ylamino)-3-nitro-chromen-2-one with a comparison to antibacterial activity of standard drugs

Nucleophilic substitution of 4-chloro-3-nitro-2-oxo-2H-chromen-2-one 3 by 6-chloro-7H-purin-2-ylamine (a), yielded the corresponding 4-(6-chloro-7H-purin-8-ylamino)-3-nitro-chromen-2-one 4. The chemical structure of synthesized compound was characterized using IR and NMR spectra, and elemental analyses. The purified synthesized compound 4 was tested at concentrations 1, 3, 5 mg/ml for its antibacterial activity against three bacterial cultures: Staphylococcus aureus, Escherichia coli, and Bacillus cereus. The antibacterial activity of synthesized compound was compared to antibacterial activity of standard antibiotics cephalexine and streptomycin.     

矿物尘粒与人体正常细菌相互作用的机制

研究了4种天然及4种人工硅酸盐尘粒与人体的正常细菌大肠埃希菌及表皮葡萄球菌相互作用的机制.测定了矿物粉尘与大肠埃希菌及表皮葡萄球菌作用后的细菌数量、葡萄糖(GLU)消耗量及14种主要阳离子的浓度变化.结果表明,各种粉尘的主要化学组成各不相同,含钙的水泥尘粒以及含镁高的水镁石均能引起大肠埃希菌及表皮葡萄球菌的细菌数量、葡萄糖消耗量增加;各矿物粉尘均不能引起培养基Ti⁴⁺、Pb²⁺、Zn²⁺、K+、Na+、Ba²⁺、P⁵⁺浓度的变化,而能引起Mn²⁺、Ca²⁺、Mg²⁺、Si⁴⁺、Fe³⁺、Al³⁺、Ni³⁺浓度不同的变化;含Ca²⁺或Mg²⁺高的碱性矿物尘粒能促进细菌的生长代谢.     

产1,3-丙二醇温控重组大肠杆菌JM109(pBV220-yqhD-dhaB)的构建

1,3-丙二醇(1,3-PD)是一种重要的化工原料,其最重要的用途是作为合成聚酯PTT的单体.由于微生物发酵法生产1,3-PD具有操作简单,不易产生有毒副产物等特点,已得到广泛关注.本研究在前期工作的基础上,分别获得了来源于肺炎克雷伯氏菌的甘油脱水酶编码基因dhaB和来源于大肠杆菌的1,3-PD氧化还原酶同工酶编码基因yqhD,利用温控表达载体pBV220串联构建了重组质粒pBV220-yqhD-dhaB,将其转化大肠杆菌得到产1,3-丙二醇温控重组大肠杆菌JM109(pBV220-yqhD-dhaB).该重组菌在LB培养基中,30℃好氧培养12 h至对数生长中期,再经42℃好氧诱导发酵4 h,测得胞内甘油脱水酶和1,3-丙二醇氧化还原酶同工酶的酶活力分别达到260 U/mg蛋白和140U/mg蛋白;在含甘油40 g/L的发酵培养基中,30℃好氧培养12 h至对数生长中期,再经42℃好氧诱导发酵4 h,测得发酵液中1,3-PD含量为8.5 g/L.这将为进一步构建基因工程菌生产1,3-PD打下坚实的基础.图6表1参18     

矿物微尘对人体肠道大肠杆菌和嗜酸乳杆菌的体外作用研究

研究了采集自居住环境的矿物微尘对人体肠道菌群主要代表性菌株大肠杆菌(Escherichia coli)和嗜酸乳杆菌 (Lactobacillus acidophilus)的体外作用.测定了矿物微尘对培养基pH值的影响,微尘与菌体共同作用后培养基pH值的变化,并用活菌计数法检测了矿物微尘对大肠杆菌和嗜酸乳杆菌生长的影响.结果表明微尘对大肠杆菌的生长起抑制作用.培养基中矿物微尘质量分数达3%时即对大肠杆菌生长有显著抑制作用,微尘质量分数达20%时,大肠杆菌生长受到完全抑制.微尘质量分数在3%～20%这一较宽幅度内,大肠杆菌虽受抑制但能生长,说明大肠杆菌耐受矿物微尘影响的能力较强.矿物微尘对嗜酸乳杆菌生长影响复杂,培养基中矿物微尘质量分数低时起促进作用,质量分数较高时起抑制作用.微尘质量分数达5%时,嗜酸乳杆菌生长即受到完全抑制.嗜酸乳杆菌耐受矿物微尘影响的能力较弱,远小于大肠杆菌.结果还表明矿物微尘影响大肠杆菌与嗜酸乳杆菌生长的主要因素是微尘本身的化学成分与物理性质,微尘的碱性因素对大肠杆菌和嗜酸乳杆菌有一定影响,但不是主要影响因素.