Effects of 50 Hz rotating magnetic field on the viability of Escherichia coli and Staphylococcus aureus

This study presents results of research on the influence of rotating magnetic field (RMF) of the induction of 30?mT and the frequency of 50?Hz on the growth dynamics and cell metabolic activity of E. coli and S. aureus, depending on the exposure time. The studies showed that the RMF caused an increase in the growth and cell metabolic activity of all the analyzed bacterial strains, especially in the time interval t?=?30 to 150?min. However, it was also found that the optical density and cell metabolic activity after exposition to RMF were significantly higher in S. aureus cultures. In turn, the study of growth dynamics, revealed a rapid and a significant decrease in these values from t?=?90?min) in the case of E. coli samples. The obtained results prove that RMF (B?=?30?mT, f?=?50?Hz) has a stimulatory effect on the growth and metabolic activity of E. coli and S. aureus. Furthermore, taking into account the time of exposure, stronger influence of RMF on the viability was observed in S. aureus cultures, which may indicate that this effect depends on the shape of the exposed cells.     

Y1转座酶关联转座子在大肠杆菌和沙门氏菌基因组中的分布和结构特征

Y1转座酶关联转座子(Y1ATs)的活性催化位点为一个酪氨酸,能够切割和连接单链DNA,在原核生物分布广泛。为探究Y1转座酶关联转座子在大肠杆菌(Escherichia coli, E. coli)与沙门氏菌(Salmonella enterica, S. ente)基因组中系统进化特性,通过Hmmsearch程序对Y1转座酶关联转座子进行了挖掘分析。结果表明,Y1转座酶关联转座子广泛分布于96.84%大肠杆菌基因组和80.4%沙门氏菌基因组。根据序列比对和蛋白结构域预测将Y1转座酶关联转座子分为10类,均隶属于IS200/IS605超家族,其中11 645个属于IS200家族,4 811个属于IS605家族。IS200家族广泛分布于S. ente基因组中(72.24%),而IS605家族广泛分布于E. coli基因组中(89.38%)。IS200拷贝数以及完整拷贝数显著高于IS605。IS200家族仅含有一个Y1转座酶编码区,而IS605家族含两个开放阅读框,分别编码Y1转座酶和TnpB蛋白。IS200家族的Y1氨基酸序列高度保守(95.3%),而IS605家族的Y1和TnpB具有较高遗传多样性,为研究转座子在原核生物的遗传进化模式提供重要参考。IS200家族具有高度保守的Y1转座酶,且完整拷贝数比例较高,提示该类转座子可能具有转座活性,对其活性的挖掘有利于研制转座子介导的新型高效基因编辑工具。     

Improved heterologous production of the nonribosomal peptide‐polyketide siderophore yersiniabactin through metabolic engineering and induction optimization

Biosynthesis of complex natural products like polyketides and nonribosomal peptides using Escherichia coli as a heterologous host provides an opportunity to access these molecules. The value in doing so stems from the fact that many compounds hold some therapeutic or other beneficial property and their original production hosts are intractable for a variety of reasons. In this work, metabolic engineering and induction variable optimization were used to increase production of the polyketide‐nonribosomal peptide compound yersiniabactin, a siderophore that has been utilized to selectively remove metals from various solid and aqueous samples. Specifically, several precursor substrate support pathways were altered through gene expression and exogenous supplementation in order to boost production of the final compound. The gene expression induction process was also analyzed to identify the temperatures and inducer concentrations resulting in highest final production levels. When combined, yersiniabactin production was extended to ～175 mg L^?1. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1412–1417, 2016     

Etude au microscope électronique à balayage de la morphologie des mandibules chez les Cladocères marins

The mandibles of the parthenogenetic females of the marine calyptomeran and gymnomeran Cladocera have been examined by means of the scanning electron microscope. These investigations reveal great differences beween Calyptomera and Gymnomera with respectively pulverizing and cutting mechanisms. The appendages of the Gymnomera are morphologically very similar and these species seem able to deal with a wide variety of food.

     

中国家蚕抗菌肽ABP-CM4在E.coli中的融合表达及活性分析

参照天然抗菌肽CM4(ABP-CM4)氨基酸序列和大肠杆菌偏爱密码子,采用rPCR法获得CM4基因后重组到表达载体pET32a上,在E.coli中融合表达。表达产物以可溶性存在,经Ni2 -NTA琼脂糖亲和层析获得融合蛋白,再经甲酸切割、亲和层析和阳离子交换层析,得到纯化的重组抗菌肽。琼脂糖扩散法和液相测定法证明了纯化的抗菌肽具有抗菌活性。     

一个新的大肠杆菌单链质粒载体系统

近年来,在重组DNA领域里的一个重要进步,是发展了质粒载体系统。为不同目的建立了不同的载体。噬菌体M13载体系统的建立,大大简化了应用双脱氧末端终止法分析DNA序列和进行特异性点变异的程序。但M13载体仍有局限性,如插入较大的片段常导致载体系统的不稳定,又如操作中常须反复克隆等。许多研究报告指出,将含噬菌体M13或f1复制     

非典型性肺炎病毒S蛋白受体结合域的可溶性表达

试验目的是获得S蛋白受体结合域基因,并获得其高效表达,为SARS病毒受体的进一步研究奠定一定的基础。首先通过P（取方法获得了S蛋白起始密码和SalⅠ限制性内切酶之间包含受体结合区的片段,然后将该基因定向克隆到pET-22b原核表达载体,构建了per—22b—S1重组质粒,转化大肠杆菌并诱导目的蛋白的表达,经SDS—PAGE没有发现明显的目的蛋白带。利用载体和S1基因上的NeoⅠ酶切位点,将S1基因的信号肽序列和部分疏水序列切掉后,构建pET—22b—SNS重组质粒。pET—22b—SNS重组质粒仍然包含受体结合域序列,并且阅读框没有改变。将pET—22b—SNS转化大肠杆菌,发现明显的目的蛋白带。Westem blot结果表明表达蛋白为SARS病毒S1蛋白的一部分。     

Viability of zoonotic pathogens Escherichia coli and Salmonella in swine manure slurries with and without a urease inhibitor and thymol

Aims: To determine the effects of urine, a urease inhibitor and/or an odour‐reducing antimicrobial compound, on zoonotic pathogens in swine manure slurries. Methods and Results: Swine faeces were collected and blended with different amounts of urine. Marker strains of Salmonella enterica subsp. enterica serovar Typhimurium and Escherichia coli O157:H7 were inoculated into the manure slurries with or without the urease inhibitor, N‐(n‐butyl) thiophosphoric triamide, or the antimicrobial compound thymol. In slurries containing approximately 1 : 1 or 2 : 1 of urine and faeces, the decreases in colony forming units (CFU) of Salm. Typhimurium and E. coli were similar and few counts were observed after 14 days. When the urine content of the slurry was increased to 5 : 1, both strains died off rapidly. When NBPT was added to the slurries, pathogen CFU was not affected in the 1 : 1 slurry. The 2 : 1 and 5 : 1 slurries had higher urea content and NBPT increased CFU of both pathogens. Thymol addition rapidly killed the pathogens and few CFU of Salmonella or E. coli were observed after day 1. Conclusions: These experiments demonstrate that urea hydrolysis in swine manure affects pathogens. Inhibition of urea hydrolysis may promote pathogen viability. Significance and Impact of the Study: Manure amendments that prevent urea hydrolysis may promote pathogen persistence. Additional treatments with antimicrobials may be required to kill pathogens.     

The expression of an R3 lipopolysaccharide-core by pathotypes of Escherichia coli

AIMS: To investigate the incidence of an R3 lipopolysaccharide (LPS)-core amplicon in a range of pathotypes of Escherichia coli, including Verocytotoxin-producing E. coli (VTEC), enteroaggregative E. coli (EAggEC) and enteropathogenic E. coli (EPEC). METHODS AND RESULTS: A total of 100 strains of E. coli belonging to a range of pathotypes, including 41 strains of VTEC, were screened for the genes encoding the R3 LPS-core using PCR. Fifty-four per cent produced an amplicon with the R3 primer set. Of the 41 VTEC, 66% had an R3 LPS-core with a PCR product being observed with all strains belonging to serotypes O26:H11, O111ac:H- and O145:H25. However, 46% of enteroaggregative E. coli and 50% of enteropathogenic E. coli were also shown to have an R3 LPS-core structure. CONCLUSIONS: Strains with an R3 LPS-core are widely distributed within the species E. coli. SIGNIFICANCE AND IMPACT OF THE STUDY: Strains of E. coli with an R3 LPS-core structure appear not to be associated with a specific pathotype.     

Response dynamics of 26-, 34-, 39-, 54-, and 80-kDa proteases in induced cultures of recombinant Escherichia coli

Several researchers have demonstrated that the presence of a heterologous protein in recombinant Escherichia coli elicits a response similar to the heat-shock response, which includes enhanced protease expression. The present work detects, quantifies, and characterizes intracellular protease activity in E. coli that are "shocked" by the induction of a recombinant protein, CAT, which is an endogenous protein in some E. coli strains. A novel, sodium dodecyl sulfate gelatin poly-acrylamide gel electrophoresis (SDS-GPAGE) method is used to detect, quantify, and characterize the presence of these proteases. A hypothesis is proposed which links the amplified protease activity to a temporary depletion of specific amino acid pools, and a stringent-like stress response. (c) 1993 John Wiley & Sons, Inc.     

Parametric sensitivity of stoichiometric flux balance models applied to wild-type Escherichia coli metabolism

The esterification of lauric acid with geraniol catalyzed by the commercially immobilized lipase preparation from Mucor miehei, Lipozyme(R), was studied in well-stirred flasks. The enzyme support was characterized in terms of its internal and external surface area, protein location, and protein content. It was found that the enzyme was mainly located on the external surface of the support, therefore, internal diffusional limitations were not important. It was also shown that the protein content of the support depends on the size of the particle, with smaller particles containing higher amounts of protein per unit weight. Under the conditions studied, the reaction was not under external mass transfer limitations, and the initial reaction rate depended on the size of the support particles. This was mainly due to the different protein contents on the support as a function of particle size and not to internal or external mass transfer limitations. Also, it was found that the inhibition exerted by water was predominantly a physical effect due to its accumulation around the enzyme. It was also found that the reaction was substrate inhibited by lauric acid, but not by geraniol. (c) 1995 John Wiley & Sons, Inc.     

Improved E. coli erythromycin A production through the application of metabolic and bioprocess engineering

In this report, small-scale culture and bioreactor experiments were used to compare and improve the heterologous production of the antibiotic erythromycin A across a series of engineered prototype Escherichia coli strains. The original strain, termed BAP1(pBPJW130, pBPJW144, pHZT1, pHZT2, pHZT4, pGro7), was designed to allow full erythromycin A biosynthesis from the exogenous addition of propionate. This strain was then compared against two alternatives hypothesized to increase final product titer. Strain TB3(pBPJW130, pBPJW144, pHZT1, pHZT2, pHZT4, pGro7) is a derivative of BAP1 designed to increase biosynthetic pathway carbon flow as a result of a ygfH deletion; whereas, strain TB3(pBPJW130, pBPJW144, pHZT1, pHZT2, pHZT4-2, pGro7) provided an extra copy of a key deoxysugar glycosyltransferase gene. Production was compared across the three strains with TB3(pBPJW130, pBPJW144, pHZT1, pHZT2, pHZT4, pGro7) showing significant improvement in erythronolide B (EB), 3-mycarosylerythronolide B (MEB), and erythromycin A titers. This strain was further tested in the context of batch bioreactor production experiments with time-course titers leveling at 4 mg/L, representing an approximately sevenfold increase in final erythromycin A titer.     

The structure of some cephalopod statoliths

Summary The statoliths of Sepia officinalis, Octopus vulgaris, Alloteuthis subulata and Taonius megalops have a smooth outline, but an irregular shape. They have projections and indentations. The statoliths from a pair of statocysts are usually quite similar in size and shape, and the general pattern is probably maintained throughout the size range of the species. Statoliths from large animals are marginally larger than those from smaller ones. The statolith usually occupies only a small part of the cavity of the statocyst, and it is situated in the anterior part of the statocyst. They are joined to the macula by hairs extending from it. These hairs are very delicate and easily broken during preparation of the specimens. The hairs are much longer and narrower than the receptor cilia of the macula. The receptor cilia are enclosed within holes in the tangled hairlike anchoring fibrils.The statolith is made up of crystalline subunits, the statoconia. The crystals vary in size, they are usually elongated, hexagonal with pointed ends. The statolith consists of a closely packed mass of these crystals, sometimes they are irregularly arranged, where in others they are stacked with their long axes parallel. In Sepia officinalis and Taonius megalops, the crystals are arranged in regular shaped packets and these packets of crystals are stacked together. These larger subunits are not always arranged in a regular way, and their major axes can be organised in several different ways. The size and outline of these large subunits do vary in different parts of the statolith.The external surface of the statolith is macroscopically smooth. Over some parts there is a surface layer covering the rod-like crystals that make up the major bulk of the stone. In other regions, the surface is rough at a microscopic level, the roughness is produced by the exposed ends of the filamentous crystals. The crystals are composed of calcium carbonate in the form of aragonite.I wish to thank Professor J.Z. Young, FRS, for considerable help, advice and encouragement throughout this study. Dr. A. Boyde generously allowed me to use his scanning electron microscope and gave freely of his expertise and time. Dr. J. Fitch kindly gave me some fossil statoliths and Dr. J. Elliott examined them with his x-ray diffraction apparatus. Dr. Marion Nixon helped me to collect and prepare the specimens. Mrs. E. Bailey, Miss P. Stephens and Mr. R. Moss provided the expert technical assistance     

大肠杆菌半乳糖结合凝集素的提取纯化

采用琼脂糖凝胶CL-6B（Sepharose CL-6B）亲和层析以及Sephadex G-75凝胶分子筛等对大肠杆菌（Esche-richia coli,E.coli）半乳糖凝集素进行了纯化。结果显示,目标蛋白经简单的步骤即可以得到纯化,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）及凝血实验证明纯化蛋白为E.coli半乳糖凝集素,蛋白提取回收率为11.4%。研究首次从E.coli蛋白提取液中分离得到纯的半乳糖凝集素,且此方法简单快捷,优越性明显。应用此方法将有利于微生物半乳糖凝集素的深入研究。