Characterization of Extracellular Polymeric Substances Produced by Pseudomonas fragi Under Air and Modified Atmosphere Packaging

Extracellular polymeric substances (EPS) play an important role in bacterial biochemical properties. The characteristics of EPS from 2 strains of Pseudomonas fragi cultured in meat aerobically (control) and in modified atmosphere packaging (MAP) were studied. The amount and components of EPS, the surface properties, and the effect on biofilm formation of several spoilage organisms were evaluated. The results showed that MAP inhibited the growth of the P. fragi strains. Compared with the control, more loose and less bound EPS (containing protein and carbohydrate) were produced by P. fragi in MAP samples. MAP also caused increased cell autoaggregation and surface hydrophobicity. After the removal of the EPS, the surface property changes were strain‐dependent, suggesting that membrane compositions were also changed. In addition, the EPS displayed significant antibiofilm activity on Pseudomonas fluorescens and Serratia liquefaciens. In conclusion, P. fragi strains not only modified the amount, components, and surface properties of EPS but also changed the cell membrane compositions to adapt to MAP stress. Moreover, EPS may play an important role in microbial community competitions.     

Characterization of Phenol Metabolization by P. stutzeri N2

Characterization of phenol metabolization by P. stutzeri N2 was proposed based on the identification of relevant metabolites and enzyme activities of catechol 1,2-oxygenase and catechol 2,3-dioxygenase. P. stutzeri N2 started to grow on 400 mg l^?1 phenol with no obvious lag phase, and then phenol decomposition was rapidly completed within 32 h. Results of enzyme activities, UV spectrums of cultures, and the mass spectrum identification of cis, cis-muconic and 2-HMS, which showed that catechol C12O and C23O were acting simultaneously on the fission of the phenol ring. The existence of P-hydroxybenzoic acid and straight-chain saturated dicarboxylic acids in the culture medium showed that carboxylation and hydrogenation were involved during the phenol degradation. Metabolites containing five or four carbons indicated that catechol and its immediate ring-cleavage intermediates could be furthermore metabolized by P. Stutzeri N2. Therefore, P. stutzeri N2 which could degrade phenol with high efficiency and diverse pathways could have potential applications in the treatment of wastewater containing phenol.     

Achieving the Best Yield in Glycolipid Biosurfactant Preparation by Selecting the Proper Carbon/Nitrogen Ratio

Pseudomonas aeruginosa RS29, the native biosurfactant-producing strain isolated from the oil fields of Assam, India was used to investigate the influence of the carbon nitrogen ratio on production of the biosurfactant. The biosurfactant producing ability of the strain was measured based on surface tension (ST) reduction of the culture medium and the emulsification (E24) index. Production was greatly influenced by the sources of nitrogen and carbon as well as the carbon to nitrogen (C/N) ratio. Sodium nitrate was the best nitrogen source and the water miscible carbon source, glycerol was observed as the best carbon source for maximum biosurfactant production. The C/N ratio 12.5 allowed the maximum production of biosurfactant by the RS29 strain. At this C/N ratio, 55 % ST of the culture medium was reduced by the produced biosurfactant. Concentrations of crude and rhamnolipid biosurfactant obtained at this particular C/N ratio were 5.6 and 0.8 g/l respectively. The RS29 strain was novel as it was able to produce a sufficient amount of biosurfactant utilizing a much lower amount of the water miscible carbon source, glycerol. Extraction of the biosurfactant by a chloroform–methanol (2:1) mixture was the best method to obtain the highest biosurfactant from the culture medium of the strain. The biosurfactant was confirmed as a mixture of mono and di-rhamnolipid congeners, Rha–C₁₀–C₁₀–CH₃ being the most abundant one. The biosurfactant was a good foaming and emulsifying agent.     

Susceptibility of Mycobacterium immunogenum and Pseudomonas fluorescens to Formaldehyde and Non-Formaldehyde Biocides in Semi-Synthetic Metalworking Fluids

Mycobacterium immunogenum, a newly identified member of the Mycobacterium chelonae_M. abscessus complex is considered a potential etiological agent for hypersensitivity pneumonitis (HP) in machine workers exposed to contaminated metalworking fluid (MWF). This study investigated the biocidal efficacy of the frequently applied commercial formaldehyde-releasing (HCHO) biocides Grotan and Bioban CS 1135 and non-HCHO type biocides Kathon 886 MW (isothiazolone) and Preventol CMK 40 (phenolic) toward this emerging mycobacterial species (M. immunogenum) in HP-linked MWFs, alone and in presence of a representative of the Gram-negative bacterial contaminants, Pseudomonas fluorescens, using two semi-synthetic MWF matrices (designated Fluid A and Fluid B). Relative biocide susceptibility analysis indicated M immunogenum to be comparatively more resistant (2-1600 fold) than P. fluorescens to the tested biocides under the varied test conditions. In terms of minimum inhibitory concentration, Kathon was the most effective biocide against M. immunogenum. Fluid factors had a major effect on the biocide susceptibility. Fluid A formulation provided greater protective advantage to the test organisms than Fluid B. Fluid dialysis (Fluid A) led to an increased biocidal efficacy of Grotan, Kathon and Preventol against M. immunogenum further implying the role of native fluid components. Used fluid matrix, in general, increased the resistance of the two test organisms against the biocides, with certain exceptions. M. immunogenum resistance increased in presence of the co-contaminant P. fluorescens. Collectively, the results show a multifactorial nature of the biocide susceptibility of MWF-colonizing mycobacteria and highlight the importance of more rigorous efficacy testing and validation of biocides prior to and during their application in metalworking fluid operations.     

可得然多糖生产菌株的筛选、鉴定及其培养条件的研究

可得然多糖是一种可溶性多糖,是由葡萄糖残基以β-1,3糖苷键连接组成的直链无分支葡聚糖.这种多糖被广泛应用在食品行业中.聚合度的不同导致这种多糖具有不同的理化性质.因此发现具有合成可得然多糖的不同微生物资源是非常重要的,但到目前为止,只报道了粪产碱杆菌和农杆菌具有合成这种多糖的能力.研究从土壤中筛选到一株能合成可得然多糖的细菌,通过16S rDNA扩增、序列比对,发现该菌株(Cu-4)是革兰氏阳性的假单胞菌属(Pseudomonas sp.).通过对影响该菌株合成可得然多糖的因素研究,发现葡萄糖是最适碳源,酵母粉是最适氮源,培养基初始最适pH为7.0.文章首次报道了Pseudomonas sp.也能合成可得然多糖.     

门多萨假单胞菌Pseudomonas mendocina P10脂肪酶基因的克隆及其在酵母中的表达

采用同源克隆法获得了门多萨假单胞菌Pseudomonas mendocina P10脂肪酶基因的全长序列,DNA测序结果表明,该基因的DNA全长序列为801bp,编码267个氨基酸,推测蛋白相对分子量为29.95ku.将该脂肪酶基因连接到pPICZαA载体上得到重组表达质粒,转化Pichia pastoris X-33.脂肪酶基因在Pichia pasttoris X-33中实现了分泌性表达,摇瓶发酵液上清酶活最大可达8U/mL.     

石油脱硫菌Pseudomonas stutzeri UP-1的固定化研究

利用从胜利油田的土壤和污水中筛选得到的能降解二苯并噻吩(DBT)的Pseudomonas stutzeri UP-1菌株进行了固定化研究，考察了固定化条件对菌种脱硫性能的影响，对比了固定化前后该菌种的脱硫性能。结果表明。选取海藻酸钠为包埋法固定化载体，固定化最佳操作条件为4℃交联、海藻酸钠质量分数为3％、胶液体积(ml)与细菌质量(g)比为20时，固定化细胞仍然具有良好的的脱硫性能，固定化细胞的重复使用性能和使用寿命大大高于未固定化细胞，通过再生活化其使用寿命可以达到700h。