序批式生物膜法的脱氮特性研究

对序批式生物膜工艺的脱氮特性进行了探讨，并提出了过量储存-SND脱氮作用机理，即厌氧段脱氮主要靠生物膜对含碳、氮有机物的过量储存作用，而好氧段脱氮则靠生物膜的同步硝化反硝化(SND)作用，反硝化的有机碳源主要为生物膜在厌氧段过量储存的有机物。镜检发现反应器中的优势菌属为假单胞菌属，其次为气单胞菌属、芽孢杆菌属、微球菌属、硝化杆菌属。     

变形假单胞菌2-酮基葡萄糖酸激酶基因的克隆、表达及生物信息学分析

采用聚合酶链式反应技术,从2-酮基葡萄糖酸工业生产菌株变形假单胞菌JUIM01中克隆2-酮基葡萄糖酸激酶的全长基因kguK,并在此基础上构建了重组菌株大肠杆菌BL21(DE3)/pET-28a(+)-kguK。在异丙基-β-D-硫代半乳糖苷的诱导下,该重组菌株表达了一个特异的分子质量约为36.0 ku融合蛋白,且该蛋白的Western-Blot鉴定结果显示为阳性。生物信息学分析结果表明,该蛋白是一种由305个氨基酸残基组成的亲水性蛋白,定位于细胞质中,存在着与pfkB家族蛋白类似的保守结构域,其二级结构中α-螺旋、延伸链和无规则卷曲所占的比例分别为35.73%、12.79%和51.48%。     

Antagonism between jasmonate- and salicylate-mediated induced plant resistance: effects of concentration and timing of elicitation on defense-related proteins,herbivore, and pathogen performance in tomato

The jasmonate (JA) and salicylate (SA) signaling pathways in plants provide resistance to herbivorous insects and pathogens. It is known that these pathways interact, sometimes resulting in antagonism between the pathways. We tested how the timing and concentration of elicitation of each pathway influenced the interaction between the jasmonate and salicylate pathways measured in terms of five biochemical responses and biological resistance to caterpillars and bacteria. The salicylate pathway had a stronger effect on the jasmonate pathway than did the reverse. The negative signal interaction was generated by two distinct paths in the plant. A negative interaction in the biochemical expression of the two pathways was most consistent in the simultaneous elicitation experiments compared to when the elicitors were temporally separated by two days. Herbivore bioassays with Spodoptera exigua also consistently reflected an interaction between the two pathways in the simultaneous elicitation experiments. The negative signal interaction reducing biological resistance to the herbivore was also demonstrated in some temporally separated treatment combinations where attenuation of the biochemical response was not evident. Concentration of the elicitors had an effect on the pathway interaction with consistent biochemical and biological antagonism in the high concentration experiments and inconsistent antagonism in the low concentration experiments. The bacterial pathogen, Pseudomonas syringae pv. tomato (Pst), consistently showed reduced lesion development on plants with SA responses activated and, in some experiments, on JA-elicited plants. Resistance to Pst was not reduced or enhanced in dual-elicited plants. Thus, signal interaction is most consistent when elicitors are applied at the same time or when applied at high doses. Signal interaction affected the herbivore S. exigua, but not the pathogen Pst.     

The role of LasR active site amino acids in the interaction with the Acyl Homoserine Lactones (AHLs) analogues: A computational study

The present work combines molecular docking calculations, 3D-QSAR, molecular dynamics simulations and free binding energy calculations (MM/PBSA and MM/GBSA) in a set of 28 structural analogues of acyl homoserine lactones with Quorum Sensing antagonist activity. The aim of this work is to understand how ligand binds and is affected by the molecular microenvironment in the active site of the LasR receptor for pseudomonas aeruginosa. We also study the stability of the interaction to find key structural characteristics that explain the antagonist activities of this set of ligands. This information is relevant for the rational modification or design of molecules and their identification as powerful LasR modulators.The analysis of molecular docking simulations shows that the 28 analogues have a similar binding mode compared to the native ligand. The carbonyl groups belonging to the lactone ring and the amide group of the acyl chain are oriented towards the amino acids forming hydrogen bond like interactions. The difference in antagonist activity is due to location and orientation of the LasR side chains within the hydrophobic pocket in its binding site. Additionally, we carried out molecular dynamics simulations to understand the conformational changes in the ligand-receptor interaction and the stability of each complex. Results show a direct relationship among the interaction energies of the ligands and the activities as an antagonist of the LasR receptor.     

固定化脂肪酶生物反应器催化合成乙酸正己酯

为提高非水相酶促合成食品添加剂乙酸正己酯的效率,以羧甲基纤维素为稳定剂,通过物理吸附,将假单胞菌脂肪酶Pseudomonas cepacia lipase固定在锥形瓶内壁上,制备成简易的生物反应器,并研究了其用于催化合成乙酸正己酯的动力学。结果表明,在反应器中,加入己醇与乙酸乙烯酯的混合液,并在37℃、150r/min下摇动,反应即开始;反应液倒出,则反应停止,无需过滤。反应7h,相对于酶粉,反应器中的固定化酶,可使反应转化率提高7倍,48h后,转化率达99%。经10次循环催化、共10d与有机相的接触,仍保留有52%的转酯活性;同样条件下,不添加羧甲基纤维素的固定化酶,仅有12.8%的活性。可见,这种反应器操作方便,能有效提高酶的非水活性和稳定性,适于催化非水相反应。      

施氏假单胞菌UP-1降解二苯并噻吩的动力学模型

对施氏假单胞菌(UP-1)降解二苯并噻吩（DBT）的过程进行了宏观动力学研究,以Michaelis-Menten方程为基础确定了UP-1降解DBT的动力学模型。计算得到动力学参数vmax=84.75 mg/(l·h), Km=759.59 mg/l。通过相关指数R2和F值检验,该动力学模型高度显著,两个重要参数取值可信。在考察的底物浓度范围内菌株UP-1降解DBT过程不存在底物抑制作用,但存在产物抑制作用。求得的UP-1休止细胞降解DBT的最大降解速率为88.5 mmol/(kg DC·h),其降解脱硫能力优异。     

绿脓杆菌外膜蛋白对感染的保护作用

目的 寻找一种防治绿脓杆菌感染的有效方法。方法 用绿脓杆菌外膜蛋白免疫的兔抗血清对4种不同血清型的菌株和1株临床绿脓杆菌进行体内外交叉保护性试验。结果 家兔免疫1周后即产生抗体,并逐渐升高,至第6周抗体滴度达到1:163 840,并维持高水平;抗血清能与4种不同血清型菌株和1株临床菌株产生直接凝集反应。第8周的血清(1:8以上)与其外胰蛋白的ELISA抗体效价相近,小鼠的半保护剂量均在0.05～0.11ml之间。结论 外膜蛋白疫苗具有较强的抗原性,而且具有良好的交叉保护作用。     

Bacterial-based additives for the production of artificial snow: What are the risks to human health?

For around two decades, artificial snow has been used by numerous winter sports resorts to ensure good snow cover at low altitude areas or more generally, to lengthen the skiing season. Biological additives derived from certain bacteria are regularly used to make artificial snow. However, the use of these additives has raised doubts concerning the potential impact on human health and the environment. In this context, the French health authorities have requested the French Agency for Environmental and Occupational Health Safety (Afsset) to assess the health risks resulting from the use of such additives. The health risk assessment was based on a review of the scientific literature, supplemented by professional consultations and expertise. Biological or chemical hazards from additives derived from the ice nucleation active bacterium Pseudomonas syringae were characterised. Potential health hazards to humans were considered in terms of infectious, toxic and allergenic capacities with respect to human populations liable to be exposed and the means of possible exposure. Taking into account these data, a qualitative risk assessment was carried out, according to four exposure scenarios, involving the different populations exposed, and the conditions and routes of exposure. It was concluded that certain health risks can exist for specific categories of professional workers (mainly snowmakers during additive mixing and dilution tank cleaning steps, with risks estimated to be negligible to low if workers comply with safety precautions). P. syringae does not present any pathogenic capacity to humans and that the level of its endotoxins found in artificial snow do not represent a danger beyond that of exposure to P. syringae endotoxins naturally present in snow. However, the risk of possible allergy in some particularly sensitive individuals cannot be excluded. Another important conclusion of this study concerns use of poor microbiological water quality to make artificial snow.     

脱氮假单胞杆菌耗氧合成V_(B_(12))发酵条件的研究

对脱氮假单胞杆菌(Pseudomonas denitri ficans)耗氧合成V_(B_(12))的培养模式研究表明,分批补料发酵比分批发酵合成V_(B_(12))提高71.42%;其中液糖补料与糖蜜补料在同等条件下,产物浓度前者比后者高出19.08%。最适V_(B_(12))合成的培养条件如种龄、接种量和初始pH值分别为18h,10%和7.0;发酵过程中摇床的转速对V_(B_(12))的合成影响显著,其中最佳转速为260r/min。在以上优化条件下,V_(B_(12))的最高单位能达到140 g/L。     

Bioproduction of cis‐(1S,2R)‐indandiol,a chiral pharmaceutical intermediate,using a solid–liquid two‐phase partitioning bioreactor for enhanced removal of inhibitors

BACKGROUND: A solid‐liquid two‐phase partitioning bioreactor (TPPB) was used in the biotransformation of indene to cis‐(1S,2R)‐indandiol by Pseudomonas putida 421‐5 (ATCC 55687). Metered substrate feeding in single‐phase operation, or delivery from an immiscible liquid, have previously been employed to regulate the exposure of the biocatalyst to inhibitory concentrations of the substrate. In contrast, the solid‐liquid platform provided in situ substrate addition (ISSA) as well as simultaneous it in situ product removal (ISPR) as a means of overcoming substrate and product toxicity. Three different modes of operation were compared for their effects on the performance of this biotransformation: single‐phase, fed‐batch operation was carried out as a benchmark in 2.75 L aqueous medium, and subsequently with the inclusion of either 700 g liquid silicone oil or 700 g solid polymer beads. RESULTS: Biphasic modes achieved a 3‐fold productivity improvement with respect to single‐phase (30 to 90 mg L^?1 h^?1), and solid‐liquid productivity was similar to liquid‐liquid operation while achieving more extensive removal of inhibitory compounds resulting in a slightly higher product titer (1.29 vs 1.16 g L^?1). The operability of the reactor was improved by the phase stability of the solid polymer beads relative to immiscible organic solvents, preventing emulsion formation and facilitating analytics. CONCLUSION: Solid polymer beads replaced the immiscible liquid auxiliary phase for substrate delivery while performing simultaneous inhibitory molecule sequestration. Copyright © 2011 Society of Chemical Industry