海因酶产生菌的反应条件研究

利用恶臭假单胞菌(Pseudomonas putida)作为酶源,研究海因酶催化DL-对羟基苯海因合成N-氨基甲酰-D-对羟基苯甘氨酸,以制备D-对羟基苯甘氨酸,并就培养基的优化,反应条件的选择及酶活的影响作了探讨.研究表明,碱性条件下可显著增加酶反应速度,温度对提高海因酶活力有重要影响.培养初始pH值为7.0,温度28℃下培养24 h产海因酶达最高;在最适pH9.0和最适温度55℃下,海因酶活力达到0.7 u/mL,比原基础培养条件下提高3.5倍.     

表达荧光假单胞菌鞭毛蛋白工程菌的构建及其对黑松的毒性

构建了组成分泌表达载体pUC18ompA,将荧光假单胞菌Pf-5鞭毛蛋白的编码基因fliC克隆到pUC18ompA构建pUC18ompA-fliC,重组质粒转化E.coliBL21(DE3)构建了工程菌。工程菌与无菌松材线虫混合接种黑松无菌苗,研究其对无菌苗的致病能力。接种试验结果表明,工程菌与无菌松材线虫混合接种同样可以引起黑松无菌苗的萎蔫。进一步验证了体内荧光假单胞菌鞭毛蛋白在松材线虫病致病过程中的作用。     

The MexJK Multidrug Efflux Pump Is Not Involved in Acquired or Intrinsic Antibiotic Resistance in Pseudomonas aeruginosa,but Modulates the Bacterial Quorum Sensing Response

Multidrug efflux pumps are critical elements in both intrinsic and acquired antibiotic resistance of bacterial populations. Consequently, most studies regarding these protein machineries focus on this specific phenotype. Nevertheless, different works show that efflux pumps participate in other aspects of bacterial physiology too. Herein, we study the Pseudomonas aeruginosa multidrug efflux pump MexJK. Previous studies, using model strains lacking MexAB-OprM and MexCD-OprJ efflux pumps, support that MexJK can extrude erythromycin, tetracycline, and triclosan. However, the results here reported indicate that this potential increased extrusion, in a mutant overexpressing mexJK, does not alter the antibiotics susceptibility in a wild-type genetic background where all intrinsic multidrug efflux pumps remain functional. Nevertheless, a clear impact on the quorum sensing (QS) response, mainly in the Pqs-dependent QS regulation network and in the expression of Pqs-regulated virulence factors, was observed linked to mexJK overexpression. The production of the siderophore pyoverdine strongly depended on the level of mexJK expression, suggesting that MexJK might participate in P. aeruginosa pyoverdine-dependent iron homeostasis. All in all, the results presented in the current article support that the functions of multidrug efflux pumps, as MexJK, go beyond antibiotic resistance and can modulate other relevant aspects of bacterial physiology.     

利用废弃碳资源生产鼠李糖脂的研究进展

鼠李糖脂是微生物合成的一类糖脂类生物表面活性剂,具有优异的表面活性、理化性质和广阔的应用前景。铜绿假单胞菌是鼠李糖脂生产的主要发酵菌株,能够以多种水溶性碳源或疏水性碳源为底物发酵生产鼠李糖脂。当前,废弃碳资源导致的浪费和污染问题日益严峻,基于鼠李糖脂的特性与生产菌株的底物偏好性,以废弃碳资源为原料生产鼠李糖脂具有很大的发展潜力。本文分别介绍以废油脂(含油废水、食品加工废油和餐厨废油)、含糖废弃物(含糖的食品加工副产物、含糖果蔬废弃物和木质纤维)和废塑料等高聚废弃物为原料合成鼠李糖脂的研究进展,重点总结了不同底物和生产菌株对鼠李糖脂生产的影响。通过分析废弃碳资源当前利用现状,发现繁多的种类、较低的分类程度和难降解的组分是限制以废弃碳资源作为底物生产鼠李糖脂进一步发展的主要因素,并提出以废弃碳资源生产鼠李糖脂的未来发展方向。     

气调箱贮藏对鲜切黄瓜品质的影响及对假单胞菌的抑制作用

鲜切黄瓜在国内外具有很大的消费市场,但在贮藏期间极易受微生物污染而腐败变质。变形假单胞菌是低温贮藏条件下导致鲜切黄瓜腐败的主要微生物之一。该研究以变形假单胞菌为实验菌株,接种至鲜切黄瓜表面后,探讨3%O₂、7%CO₂气调箱贮藏对菌株生长和侵染情况,对黄瓜细胞壁降解和细胞壁降解引起的生理生化变化的影响。结果表明,与对照组相比,3%O2、7%CO₂气调贮藏抑制了变形假单胞菌的生长,保持了细胞壁的完整性,延缓了原果胶的降解,降低了纤维素酶活性,抑制了β-半乳糖苷酶活性的增加和可溶性果胶的产生,从而保持了硬度,延缓了丙二醛含量和相对电导率的增加,维持了鲜切黄瓜的外观和色泽。总之,3%O₂、7%CO₂气调箱贮藏抑制了由变形假单胞菌导致的鲜切黄瓜细胞壁的降解,延缓了腐败变质。     

Pseudomonas aeruginosa DnaK Stimulates the Production of Pentraxin 3 via TLR4-Dependent NF-κB and ERK Signaling Pathways

Microbe-derived factors trigger innate immune responses through the production of inflammatory mediators, including pentraxin 3 (PTX3). PTX3 is a soluble pattern recognition molecule that stimulates the clearance of clinically important bacterial pathogens such as Pseudomonas aeruginosa. However, the P. aeruginosa factors responsible for the production of PTX3 have not been elucidated. In this study, we found that P. aeruginosa DnaK, a homolog of heat shock protein 70, induced PTX3 production. Induction was mediated by intracellular signals transmitted through the Toll-like receptor 4 (TLR4) signaling pathway. Following receptor engagement, the stimulatory signals were relayed initially through the nuclear factor kappa B (NF-κB) signaling pathway and subsequently by extracellular signal-regulated kinases (ERK), which are mitogen-activated protein kinases. However, ERK activation was negatively controlled by NF-κB, implying the existence of negative crosstalk between the NF-κB and the ERK pathways. These data suggest that P. aeruginosa DnaK acts as a pathogen-associated molecular pattern to trigger modulation of host defense responses via production of PTX3.     

Effects of Pseudomonas chlororaphis and gaseous chlorine dioxide on the survival of Salmonella enterica on tomatoes

Control of Salmonella enterica on tomatoes is important for food safety. The aim of this research was to evaluate the survival of Salmonella enterica serovars Montevideo (SM) and Typhimurium (ST) on tomatoes exposed to gaseous chlorine dioxide and Pseudomonas chlororaphis (Pc). Pc was applied to stem scars of tomatoes prior to inoculations with SM and ST. Tomatoes were treated with gaseous ClO₂ at 0.4 mg L^?1 for 2 and 4 h (90% R.H. 13 °C), respectively. At 4 h of ClO₂ treatment, SM and ST populations were reduced to 0.82 and <0.30 log CFU g^?1, respectively. Tomatoes treated with SM and ST had 5.42 and 5.37 log CFU g^?1 of Salmonella. Tomatoes treated with Pc + Salmonella count was 2.59 (treated) and 5.83 log CFU g^?1 (control). Salmonella survival was similar at 2 and 4 h of ClO₂ treatment. Application of ClO₂ and Pc may reduce contamination of tomatoes by Salmonella serovars.     

耐低温菌株的鉴定、低温生长特征及耐低温机理初步研究

该研究以分离自长白山常年积雪覆盖的苔原带土样的耐低温菌株7a-3为研究对象，采用形态观察、生理生化试验及分子生物学技术对其进行菌种鉴定，并对其生长特性及耐低温机制进行初步研究。结果表明，菌株7a-3被鉴定为莓实假单胞菌（Pseudomonas fragi），其生长温度范围为4～33 ℃，最适生长温度为30 ℃，表面活性剂不能促进该菌株在37 ℃条件下生长，但其自身低温发酵代谢物可促进该菌在37 ℃条件下生长。相对于30 ℃，4 ℃条件下，菌株7a-3细胞膜的脂肪酸种类由长链脂肪酸向短链脂肪酸转化，不饱和脂肪酸的相对含量增加，且单不饱和脂肪酸相对含量的变化较大，细胞膜较为粗糙，初步推测其耐低温能力与细胞膜不饱和脂肪酸含量增加导致细胞膜流动性增强相关。