Altered expression of polyhydroxyalkanoate synthase gene and its effect on poly[(R)-3-hydroxybutyrate] synthesis in recombinant Escherichia coli

Polyhydroxyalkanoate (PHA) synthase (PhaC) from Wautersia eutropha was expressed in a wide range of production level in Escherichia coli XL1-Blue cells and its effects on PhaC activity, poly[(R)-3-hydroxybutyrate] [P(3HB)] production and its molecular weights were investigated. The production level of PhaC was controlled both by the amount of chemical inducer (isopropyl-β-d-thiogalactopyranoside, IPTG) added into the medium and the use of different copy number of plasmids. In a flask experiment, as PhaC production level in the cells increased, the PhaC activity also increased in the range of low PhaC concentration. However, PhaC activity did not further increase in the range of high PhaC concentration, probably due to the formation of inclusion body in the cells. The molecular weight of P(3HB) was found to decrease with increasing PhaC activity. This trend was also verified in high cell density cultivation using 10-l jar fermentor. Furthermore, we demonstrated that the use of low copy number plasmid and appropriate induction of PhaC expression were effective in achieving both high productivity and high molecular weight of P(3HB).     

免疫珠技术快速检测腹泻性大肠杆菌

采用表面光滑玻璃珠作为载体材料，用1：40的腹泻性大肠杆菌诊断血清进行包被。可制成免疫珠。这种免疫珠能与腹泻性大肠杆菌发生特异性结合。经乳糖蛋白胨培养液培养，然后根据其产酸产气进行判断。能在18h内完成腹泻性大肠杆菌的检测。其灵敏度为样液中含有腹泻性大肠杆菌数必须大于10CFu／mL．该法简单、快速、易操作、灵敏度高。     

Total surface areas of Group IVA organometallic compounds: Predictors of toxicity to algae and bacteria

There is a high correlation between molecular surface area (TSA) of triorganotin and triorganolead compounds and their toxicity towards a bacterium (Escherichia coli) and an alga (Selenastrum capricornutum). Parallel attempts to correlate other Group IVA organometals incorporating silicon or germanium were unsuccessful. It was further demonstrated, however, that a high correlation was obtainable between certain series of compounds with the same organic substituent but different metal centers involving all Group IVA elements. In both instances, the inability to obtain a quantitative structure-activity relationship (QSAR) for all systems studied appears to be a function of the solubility of the compounds. While organotin TSA values have been found to correlate well with their toxicities toward various organisms, this study clearly suggests that this type of QSAR can be readily extended to include other organometal systems, provided that there is no solubility problem and the toxicity is a function of the hydrophobicity of the organometal compounds.     

脉冲电磁场杀菌效应试验研究

利用自行研制的直流脉冲发生装置产生的脉冲电磁场分别对生活污水和纯种大肠埃希氏菌液进行电磁处理 ,研究其对细菌的杀菌效果及机理 .实验表明电磁脉冲对生活污水及纯种菌液中的细菌都有明显的致死作用 .实验结果表明 ,停留时间、水流速度、线圈绕组、输出功率、脉冲频率对脉冲磁场的杀菌效果具有影响作用 ,在停留时间 1 2h ,输出功率 4 6 8W ,脉冲频率 6 5kHz ,水流速度 0 .3m s时 ,生活污水的细菌总数从 3.9× 1 0 7个 mL下降到 1 .5× 1 0 2 个 mL ,存活率达 1 0 - 5;大肠杆菌数从 2 .4× 1 0 6 个 mL下降到 3.6× 1 0 2 个 mL ,存活率为 1 0 - 4.在此条件下 ,四种质量浓度的纯种大肠埃希氏菌液经电磁处理后 ,其细菌的存活率均在 1 0 - 3～ 1 0 - 5.细菌细胞的扫描电镜照片表明电磁作用使细胞表面产生凹陷 ,孔洞 ,细胞质溶出现象 ,从而证实了感应电流杀菌作用机制     

毛细管电泳法快速分离和检测肠毒性大肠杆菌

 建立了快速分离和检测引起仔猪腹泻的肠毒性大肠杆菌K88、K99和 987P细菌细胞的毛细管区带电泳方法 ,并进行了腹泻仔猪粪便中肠毒性大肠杆菌的应用检测分析。结果表明 ,在电泳缓冲液为 0 0 5mol/LNa2 CO3 NaHCO3(pH 9 9)、分离电压为 1 4 1kV、检测波长为 2 1 0nm的电泳条件下 ,E coliK88、K99和 987P的细胞分别具有单一、稳定的特征谱峰 ,其保留时间的相对标准偏差RSD≤ 0 9% ;在确定的实验条件下 ,实现了腹泻仔猪粪便中肠毒性大肠杆菌的快速检测分析 ,发现将出生 5d～ 6d的腹泻仔猪粪便引入培养基中增殖后主要检出K88。     

大肠杆菌有限生长的微量热及非线性动力学研究

The finite growth of Escherichia coli was studied by using a LKB 2277 BioActivity Mollitor. We found that the finite growth is a nonliear dynamic process. The nonlinear dynamic behaviour in the finite growth process and the nonlinear dynamic models describing the process were discovered and established. The curve of logistic map corresponding to the finite growth thermogram of Escherichia coli was obtained and the nonlinear dynamic parameters were calculated by means of a computer. Moreover, we also discussed the nonlinear dynamic characters of Escherichia colt in its finite growth process.     

化学发光磁酶免疫法检测O157∶H7大肠埃希菌

O157∶H7大肠埃希菌是一种致病力非常强的微生物.近年来,陆续有其引起食物中毒事件的报道.1996年日本发生O157∶H7大肠埃希菌食物中毒的爆发流行,感染病例逾万人,死亡9例.2001年我国江苏及安徽等地也发生了肠出血性O157∶H7大肠埃希菌所引发的感染性腹泻事件.目前O157∶H7大肠埃希菌实验室常规检测方法为分离培养计数法,此法费时费力,需要几天的时间才能得到检测结果[1,2],缺乏应对突发事件的能力,因此亟需开发快速检测技术.近年来,多种微生物快速检测技术如ELISA法[3]、热解质谱测定法[4]、多聚酶链反应[5]、荧光成像技术[6]、生物发…     

Superpositioned dielectrophoresis for enhanced trapping efficiency

One of the major applications for dielectrophoresis is selective trapping and fractionation of particles. If the surrounding medium is of low conductivity, the trapping force is high, but if the conductivity increases, the attraction decreases and may even become negative. However, high-conductivity media are essential when working with biological material such as living cells. In this paper, some basic calculations have been performed, and a model has been developed which employs both positive and negative dielectrophoresis in a channel with interdigitated electrodes. The finite element method was utilized to predict the trajectories of Escherichia coli bacteria in the superpositioned electrical fields. It is shown that a drastic improvement of trapping efficiency can be obtained in this way, when a high conductivity medium is employed.     

Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyalkanoates) by metabolically engineered Escherichia coli strains

Biosynthesis of polyhydroxyalkanoates (PHAs) consisting of 3-hydroxyalkanoates (3HAs) of 4 to 10 carbon atoms was examined in metabolically engineered Escherichia coli strains. When the fadA and/or fadB mutant E. coli strains harboring the plasmid containing the Pseudomonas sp. 61-3 phaC2 gene and the Ralstonia eutropha phaAB genes were cultured in Luria-Bertani (LB) medium supplemented with 2 g/L of sodium decanoate, all the recombinant E. coli strains synthesized PHAs consisting of C4, C6, C8, and C10 monomer units. The monomer composition of PHA was dependent on the E. coli strain used. When the fadA mutant E. coli was employed, PHA containing up to 63 mol% of 3-hydroyhexanoate was produced. In fadB and fadAB mutant E. coli strains, 3-hydroxybutyrate (3HB) was efficiently incorporated into PHA up to 86 mol%. Cultivation of recombinant fadA and/or fadB mutant E. coli strains in LB medium containing 10 g/L of sodium gluconate and 2 g/L of sodium decanoate resulted in the production of PHA copolymer containing a very high fraction of 3HB up to 95 mol%. Since the material properties of PHA copolymer consisting of a large fraction of 3HB and a small fraction of medium-chain-length 3HA are similar to those of low-density polyethylene, recombinant E. coli strains constructed in this study should be useful for the production of PHAs suitable for various commercial applications.