Simultaneous degradation of nitroaromatic compounds TNT,RDX, atrazine,and simazine by Pseudomonas putida HK‐6 in bench‐scale bioreactors

BACKGROUND: Environmental contamination by nitroaromatic compounds such as 2,4,6‐trinitrotoluene (TNT), hexahydro‐1,3,5‐trinitro‐1,3,5‐s‐triazine (RDX), atrazine, and/or simazine (TRAS) generated as waste from military and agricultural activities is a serious worldwide problem. Microbiological treatment of these compounds is an attractive method because many explosives and herbicides are biodegradable and the process can be made cost‐effective. We explored the feasibility of using cultures of Pseudomonas putida HK‐6 for simultaneous degradation of TRAS with the aim of microbial application in wastewater treatment in bench‐scale bioreactors. RESULTS: Experiments were conducted to study the effects of supplemental carbons, nitrogens, and Tween‐80 on the degradation of Ps. putida HK‐6 in media containing TRAS as target substrate(s). The most effective TRAS degradation was shown in the presence of molasses. Addition of nitrogen sources produced a delayed effect for the target substrate(s). Tween‐80 enhanced the degradation of target substrate(s). Simultaneous degradation of these compounds proceeded to completion within the given period. CONCLUSIONS: Ps. putida HK‐6 was capable of growth with TRAS, and the effects of supplements on TRAS degradation and simultaneous TRAS degradation were evaluated in bench‐scale bioreactors. The results of this study have practical applications in the processes of industrial waste stream treatment where the disposal of TRAS may be problematic. Copyright © 2008 Society of Chemical Industry     

Surface tension as a limiting factor for aerobic n-alkane biodegradation

A new mathematical model for n-alkane biodegradation in crude oil, heavy oil and paraffinic mixtures is described. The pattern of n-alkane degradation as a function of the inverse of hydrocarbon chain length reported in this paper can be considered as general behaviour for many aerobic n-alkane biodegradation processes. A new interpretation of n-alkane biodegradation as a function of surface tension, is given. A mathematical expression was obtained starting from the degradation values of n-alkane and relative surface tension, which is a parameter independent of fermentation conditions. An interesting parameter, b, was identified which represented the accelerating conversion factor for n-alkane biodegradation. The findings suggested that the n-alkane biodegradation. The findings suggested that he n-alkane biodegradation rate may be affected by the fermentation condition (agitation, aeration, etc.) and by the strain of microorganism, while the behaviour pattern of n-alkane degradation was essentially linked to the substrate characteristics (molecular structure, molecular weight and density).     

Microstructured Lipid Carriers (MLC) Based on N-Acetylcysteine and Chitosan Preventing Pseudomonas aeruginosa Biofilm

The aim of this work was the development of microstructured lipid carriers (MLC) based on chitosan (CH) and containing N-acetylcysteine (NAC), a mucolytic and antioxidant agent, to inhibit the formation of Pseudomonas aeruginosa biofilm. MLC were prepared using the high shear homogenization technique. The MLC were characterized for morphology, particle size, Z potential, encapsulation efficiency and drug release. The antioxidant properties of NAC-loaded microstructured carriers were evaluated through an in vitro spectrophotometer assay. Finally, the activity of NAC-CH-MLC on biofilm production by Pseudomonas aeruginosa was also evaluated. Results obtained from this study highlighted that the use of chitosan into the inner aqueous phase permitted to obtain microstructured particles with a narrow size range and with good encapsulation efficiency. NAC-loaded MLC showed higher antioxidant activity than the free molecule, demonstrating how encapsulation increases the antioxidant effect of the molecule. Furthermore, the reduction of biofilm growth resulted extremely high with MLC being 64.74% ± 6.2% and 83.74% ± 9.95%, respectively, at 0.5 mg/mL and 2 mg/mL. In conclusion, this work represents a favorable technological strategy against diseases in which bacterial biofilm is relevant, such as cystic fibrosis.     

Analysis of the Structure and Biosynthesis of the Lipopolysaccharide Core Oligosaccharide of Pseudomonas syringae pv. tomato DC3000

Lipopolysaccharide (LPS), the major component of the outer membrane of Gram-negative bacteria, is important for bacterial viability in general and host–pathogen interactions in particular. Negative charges at its core oligosaccharide (core-OS) contribute to membrane integrity through bridging interactions with divalent cations. The molecular structure and synthesis of the core-OS have been resolved in various bacteria including the mammalian pathogen Pseudomonas aeruginosa. A few core-OS structures of plant-associated Pseudomonas strains have been solved to date, but the genetic components of the underlying biosynthesis remained unclear. We conducted a comparative genome analysis of the core-OS gene cluster in Pseudomonas syringae pv. tomato (Pst) DC3000, a widely used model pathogen in plant–microbe interactions, within the P. syringae species complex and to other plant-associated Pseudomonas strains. Our results suggest a genetic and structural conservation of the inner core-OS but variation in outer core-OS composition within the P. syringae species complex. Structural analysis of the core-OS of Pst DC3000 shows an uncommonly high phosphorylation and presence of an O-acetylated sugar. Finally, we combined the results of our genomic survey with available structure information to estimate the core-OS composition of other Pseudomonas species.     

重组铜绿假单胞菌外毒素A工程菌发酵及表达产物的纯化

目的建立稳定、高效表达重组铜绿假单胞菌外毒素A(rEPA)的工程菌发酵及表达产物纯化工艺。方法规模化发酵表达rEPA的重组大肠杆菌rPE553D,离心收集菌体,渗透压调解使菌体周质间隙蛋白释放后,高速离心收集蛋白溶液。经DEAESepharoseFF、PhenylSepharose6FF疏水层析和SOURCE30Q强离子交换层析,超滤浓缩纯化rEPA。用HPLC、SDS-PAGE和Westernblot等方法检定生化和免疫学特性,并用小鼠和Vero细胞检定毒性。结果每55L培养基中rEPA产量超过4g,纯度在95%以上,细胞毒性降低了至少32000倍。其余各项指标均符合《中国生物制品规程》要求。结论已建立了收率高、纯度好、稳定、适合规模化生产rEPA的工艺。     

荧光假单胞菌RB5产嗜铁素的发酵条件

荧光假单胞菌RB5是从土壤中分离得到的一株对禾谷丝核菌有拮抗作用的细菌.通过CAS法检测分析,发现RB5菌株能产生嗜铁素.为明确荧光假单胞菌RB5产嗜铁素的发酵条件,采用摇瓶培养发酵,利用特异光谱吸收法,研究了不同培养条件对RB5菌株嗜铁素产量的影响.最终得出RB5产嗜铁素的最佳发酵条件:起始pH 7.0,接种量2％,瓶装量90 mL/250 mL,发酵时间48 h,Fe3+质量浓度为0.8 mg/L.抑菌试验表明,优化发酵条件后的RB5发酵液(OD400值2.35)有很显著的抑菌效果,抑制率达65.28％.     

Biodegradation of catechol by Pseudomonas fluorescens isolated from petroleum‐impacted soil

Bioremediation strategies have been applied to clean up petroleum hydrocarbon (PHC) impacted sites. Introducing PHC degrading microorganisms (bioaugmentation) and enhancing the in‐situ nutrients availability (biostimulation) are widely used strategies. These strategies can be combined to lead to a better bioremediation performance. In this work, Pseudomonas fluorescens was isolated from a PHC impacted site. Through a 2³ factorial design plan, the effect of various combinations of nitrate, sulphate, and phosphate ions on the PHC bioremediation performance by P. fluorescens was investigated using catechol, an essential metabolic intermediate of BTEX degradation, as the sole carbon source. The maximum specific catechol degradation rate was chosen as the response to evaluate the catechol bioremediation performance. The ANOVA results indicated that the presence of nitrate ions alone lowered the maximum specific catechol degradation rate, which can be explained by the accumulation of nitrites and ammonia during the denitrification process by P. fluorescens. It was noted that dosing sulphate ions alone did not affect the bioremediation performance, which indicates P. fluorescens can grow in a sulphur‐limited environment. In contrast, the presence of sulphate and nitrate ions together can lead to a higher specific catechol degradation rate. This may be caused by the presence of sulphate that can suppress the production of nitrites. The importance of phosphate ions on catechol biodegradation was investigated. The absence of phosphate led to incomplete biodegradation. Introducing phosphate ions can accelerate catechol degradation, which can be explained by the secretion of organic acids.     

低温等离子体处理模式对冷藏南美白对虾中假单胞菌的抑菌效果及机理研究

摘 要：目的 探究低温等离子体(cold plasma, CP)处理模式对冷藏南美白对虾中常见荧光假单胞菌(Pseudomonas fluorescens)的抑菌效果及其作用机制。方法 通过CP直接处理和循环处理P. fluorescens,研究了两种处理模式下臭氧含量动态变化对P. fluorescens的生长曲线、细胞活力,生物膜形成、细胞壁、细胞膜完整性和南美白对虾菌落总数及假单胞菌数等指标的影响。结果 两种处理模式在CP处理3 min或3 cycles后,包装内臭氧含量达到最高值,分别为(850±10) mg/m3和(874±20) mg/m3。CP循环处理模式使得臭氧含量随处理循环数递增,因此获得更长的臭氧存在时间从而具有更大的抑菌能力。P. fluorescens生长曲线表明CP处理使得菌体延迟期变长且对数生长期推迟。此外,CP处理后的P. fluorescens细胞活力显著下降(P<0.05),CP-1 min,CP-3 min和CP-3 cycles组的细胞活力分别为33.03%、5.90%和4.82%。同时相比CP-3 min组,CP-3 cycles组的P. fluorescens生物膜OD值下降27.61%。碱性磷酸酶(alkaline phosphatase, AKP)活性和核酸蛋白泄漏量结果表明,细胞壁和细胞膜完整性受损可能是P. fluorescens失活的直接原因。对虾保鲜测试结果证实,贮藏第6 d,CP-3 cycles组虾体中的菌落总数和假单胞菌数相比CP-3 min组分别降低了58.02%和79.54%。结论 CP循环处理模式通过延长对臭氧与对虾的暴露时间,提高了对P. fluorescens的灭活效果,同时还具有更优越的保鲜能力。本研究为开发基于CP技术的新型保鲜技术应用提供了理论参考。 关键词：低温等离子体;荧光假单胞菌;抑菌机制;保鲜     

碱性脂肪酶生产菌假单孢菌4631菌株的筛选

从３２个土样中分离获得碱性脂肪酶产生菌１６６株，其中４６３１号菌株产酶能力最高，达到９．１ＩＵ／ｍｌ。初步鉴定为假单孢菌属。对研究结果统计分析表明，Ｄ／ｄ值与ＬＡ呈线性相关关系。