Microcalorimetric Evaluation of the Effect of Kanamycin： An Analysis Based on the Median-Effect Principle

A study of the effect of drug, kanamycin, on the growth metabolism of recombinant Escherichia coli B 1 was carded out by microcalorimeter monitoring of the metabolic activity of treated cells. Power-time curves of growing recombinant Escherichia coli cell suspensions, treated with different kanamycin doses, were recorded. The extent of the effect was evaluated by changes in the slopes of the microcalorimetric curves and the kinetics of the drug action was interpreted from the time at which these changes reached their maximum values and maintained their maximum values. Experimental dose-effect relationships conform to the median-effect principle of the mass-action law： fa/（1-fa）=（D/D50）^m. A plot of y=lg[（fa）^1-1]^-1 versus x=lg D gives the slope m, D50 and R∞. The experimental results revealed that high concentration of kanamycin had an inhibitory effect on the growth of recombinant Escherichia coli B 1 in the lg phase, and had a promoting effect in the stationary period. Moreover, it was demonstrated that microcalorimetry was a reliable method for the detection of modulatory effects in biology.     

On-line sample preconcentration with chemical derivatization of bacterial biomarkers by capillary electrophoresis: a dual strategy for integrating sample pretreatment with chemical analysis

Simple, selective yet sensitive methods to quantify low-abundance bacterial biomarkers derived from complex samples are required in clinical, biological, and environmental applications. In this report, a new strategy to integrate sample pretreatment with chemical analysis is investigated using on-line preconcentration with chemical derivatization by CE and UV detection. Single-step enantioselective analysis of muramic acid (MA) and diaminopimelic acid (DAP) was achieved by CE via sample enrichment by dynamic pH junction with ortho-phthalaldehyde/N-acetyl-L-cysteine labeling directly in-capillary. The optimized method resulted in up to a 100-fold enhancement in concentration sensitivity compared to conventional off-line derivatization procedures. The method was also applied toward the detection of micromolar levels of MA and DAP excreted in the extracellular medium of Escherichia coli bacterial cell cultures. On-line preconcentration with chemical derivatization by CE represents a unique approach for conducting rapid, sensitive, and high-throughput analyses of other classes of amino acid and amino sugar metabolites with reduced sample handling, where the capillary functions simultaneously as a concentrator, microreactor, and chiral selector.     

铜和不锈钢材料抗菌生物效应的研究

采用电分析化学方法、光度分析方法研究了铜和不锈钢材料抗菌的生物效应。以大肠杆菌和发光细菌为例,不锈钢材料对它们几乎没有抗菌生物效应,但是铜材料对它们只需作用0．5h,细菌死亡数量则可达到90％以上。该结果在医药卫生消毒、罐装食品的封装及饮用水的管道输送等方面有着重要的意义。     

Biosynthesis of Poly-(3-hydroxybutyrate) under the Control of an Anaerobically Induced Promoter by Recombinant Escherichia coli from Sucrose

Poly-(3-hydroxybutyrate) (PHB) is a polyester with biodegradable and biocompatible characteristics and has many potential applications. To reduce the raw material costs and microbial energy consumption during PHB production, cheaper carbon sources such as sucrose were evaluated for the synthesis of PHB under anaerobic conditions. In this study, metabolic network analysis was conducted to construct an optimized pathway for PHB production using sucrose as the sole carbon source and to guide the gene knockout to reduce the generation of mixed acid byproducts. The plasmid pMCS-sacC was constructed to utilize sucrose as a sole carbon source, and the cascaded promoter P_3nirB was used to enhance PHB synthesis under anaerobic conditions. The mixed acid fermentation pathway was knocked out in Escherichia coli S17-1 to reduce the synthesis of byproducts. As a result, PHB yield was improved to 80% in 6.21 g/L cell dry weight by the resulted recombinant Escherichia coli in a 5 L bed fermentation, using sucrose as the sole carbon source under anaerobic conditions. As a result, the production costs of PHB will be significantly reduced.     

Hydrophobicity and the gastrointestinal tract: methods of determination, its source and implications for bacterial adherence

Increasing research efforts are now focused on characterizing the physicochemical properties of microbial pathogens and the biologic surfaces to which the organisms adhere. Interest is intense because there is the potential to develop novel strategies to disrupt the infectious process and thereby employ therapeutic agents other than antimicrobial compounds. This review will focus on the gut as a model system in order to highlight advances in the field.     

Conversion of biomass to ethanol

In theEscherichia coli cell-free system, the modification of cell extract can be achieved by preparation of the strains carrying additional property or those being induced with a certain gene expression prior to harvesting. In this study, we analyzed the cell-free system with S30 extract containing T7 RNA polymerases (S30 extract-T7pol) prepared from E.coli BL21(DE3) strain, which includes T7 RNA polymerase from extrinsic genes by IPTG induction, as a model for the improvement of the cell-free system. The fact that a significant degree of mRNA degradation was observed in the cell-free system with S30 extract-T7pol indicates the increase of ribonuclease activity was an unfavorable influence derived from the cell-extract modification process. We also showed that this influence was settled by the addition of an effective ribonuclease inhibitor, such as copper (II) ion, to the reaction mixture.     

The effect of cellular energetics on foreign protein production

Escherichia coli strain F-122 was used to determine if there are additional physiological effects, other than decreasing energetic efficiency accompanied by the excretion of the acetate, on foreign protein production. This organism was the host for expressing HIV₅₈₂-β-galactosidase fusion protein under the control of thetrp promoter, with ampicillin resistance. By comparing parallel batch cultures with and without acetate addition, it was found that the presence of acetate in the media did not influence β-galactosidase activity. In these experiments, it appears that the low protein productivity often observed during acetate formation is the result of inefficient cell metabolism, rather than acetate acting as a specific inhibitor of protein production.     

以新亚甲基蓝为电子媒介体的大肠杆菌微生物燃料电池的研究

以新亚甲基蓝(NMB) 为电子媒介体, 大肠杆菌为微生物催化剂, 设计了微生物燃料电池(MFC). 该MFC的开路电压为0.760 V, 短路电流为1.108 mA, 最大输出功率为116 mW/m2, 此时所对应的电流密度为390 mA/m2. 比较了中性红(NR)和NMB作为电子媒介体对MFC性能的影响. 实验结果表明, 以NMB为电子媒介体的MFC的开路电压比以NR为电子媒介体的MFC的开路电压低, 但其开路电压达到稳定所需要的时间更短, 而且其短路电流比后者高. 当放电电流大于114 mA/m2时, 前者比后者的输出功率高, 在负载1000 Ω放电时, 前者比后者有更好的稳定性.     

硝酸镧引起Escherichia coli B代谢过程热爆发及其机理研究

采用微量热法研究了硝酸镧对Escherichia coli B生长代谢过程的影响, 发现高浓度硝酸镧引起E. coli B热谱图出现异常变化: 生长速率常数k值增大、产热峰显著升高和总发热量异常增加. 当硝酸镧浓度为300和500 mg/L时, 培养物在培养过程的总发热量分别是正常条件下的3.89和2.54倍. 用生物学方法对细胞存活率和生物量进行测定结果表明, 细胞在高浓度硝酸镧条件下增殖受到抑制、细胞生物量减少. 表明高浓度的硝酸镧存在时, E. coli B细胞生长受到抑制反而释放出比正常生长细胞多得多的热量, 将抑制状态细胞释放大量热量的现象称为热爆发. 分析热爆发的原因, 认为是La^3＋离子破坏细胞壁外膜而增加其透性, 导致细胞膜与外膜间的质子电化学势因质子外泄而降低或者不能形成, 氧化磷酸化过程中的能量不能有效地转化为ATP, 而以热能的方式释放出来. 细胞由于缺乏生物通用能量ATP, 因而其生长受到抑制.     

大肠杆菌氧化还原辅因子代谢工程

NAD⁺、NADP⁺、NADH、NADPH等四类烟酰胺类辅因子也被称为氧化还原辅因子,是维持细胞氧化还原平衡,驱动胞内许多分解与合成反应的重要分子。近年来,辅因子在生物转化系统中的作用受到研究者的重视,氧化还原辅因子更成为人们关注的重点。文章综述了大肠杆菌（Escherichia coli, E. coli）氧化还原辅因子代谢工程研究的最新进展,重点总结最近发展的氧化还原辅因子代谢工程策略,讨论不同代谢工程策略对细胞氧化还原辅因子水平的影响及其在生物合成中的应用,并展望了辅因子代谢工程未来发展方向。