鼠李糖脂对铜绿假单胞菌在堆肥颗粒介质中吸附与传输影响的初步研究

以铜绿假单胞菌(Pseudomonasaeruginosa)AB93066及其发酵生产的鼠李糖脂(rhamnolipid)为研究对象,通过柱淋洗实验考察了不同浓度鼠李糖脂对堆肥颗粒中单一菌属铜绿假单胞菌吸附传输情况的影响.实验结果显示,一定浓度(100～400mg·L-1)的鼠李糖脂能够有效地减弱细菌在堆肥颗粒介质中的吸附作用,增强细菌在介质中的传输和分散,使细菌得以传输到样品柱的更深层.这将有利于深层堆肥有机物的生物降解.原因可能是鼠李糖脂的存在改变了菌体和堆肥颗粒的表面性质,并且使菌体和颗粒之间产生位阻效应,减弱了它们之间的亲合性.

Reuse of waste frying oil for production of rhamnolipids using Pseudomonas aeruginosa zju.u1M

In this work,rhamnolipid production was investigated using waste frying oil as the sole carbon source. By culture in shaking flasks,a naturally isolated strain synthesized rhamnolipid at concentration of 12.47 g/L and its mutant after treatment by UV light increased this productivity to 24.61 g/L. Fermentation was also conducted in a 50 L bioreactor and the productivity reached over 20 g/L. Hence,with a stable and high productive mutant strain,it could be feasible to reuse waste frying oil for rhamnolipid production on industrial scale.

Optimizing rhamnolipid production by Pseudomonas aeruginosa ATCC 9027 grown on waste frying oil using response surface method and batch-fed fermentation

Rhamnolipid production by Pseudomonas aeruginosa ATCC 9027 with waste frying oil as sole carbon source was studied using response surface method. Cultures were incubated in shaking flask with temperature, NO3- and Mg2+ concentrations as the variables. Meanwhile, fed-batch fermentation experiments were conducted. The results show that the three variables are closely related to rhamnolipid production. The optimal cultivation conditions are of 6.4 g/L NaNO3 , 3.1 g/L MgSO4 at 32 ℃, with the maximum rhamnolipid production of 6.6 g/L. The results of fed-batch fermentation experiments show that feeding the oil in two batches can enhance rhamnolipid production. The best time interval is 72 h with the maximum rhamnolipid production of 8.5 g/L. The data are potentially useful for mass production of rhamnolipid on oil waste with this bacterium.

Batch washing of saturated hydrocarbons and polycyclic aromatic hydrocarbons from crude oil contaminated soils using bio-surfactant

Desorption of total saturated fractions(i.e. SAT, defined for this study as the summation of the concentrations of the saturated hydrocarbon from n-C10 to n-C26) and polycyclic aromatic fractions(i.e. PAH, defined as the summation of the concentrations of all polycyclic aromatic fractions including the 16 EPA priority PAH) in two types of soils subjected to the changes of p H and salinity and different bio-surfactant concentrations were investigated. In general, compared with the experiments without bio-surfactant addition, adding rhamnolipid to crude oil-water systems at concentrations above its critical micelle concentration(CMC) values benefits SAT and PAH desorption. The results indicate that the change of p H could have distinct effects on rhamnolipid performance concerning its own micelle structure and soil properties. For loam soil, the adsorption of non-aqueous phase liquid(NAPL) and rhamnolipid would be the principle limiting factors during the NAPL removal procedure. For sand soil, less amount of rhamnolipid is adsorbed onto soil. Thus, with the increase of salinity, the solubilization and desorption of rhamnolipid solution are more significant. In summary, the p H and salt sensitivity of the bio-surfactant will vary according to the specific structure of the surfactant characteristics and soil properties.

Enzymatic reaction of ethanol and oleic acid by lipase and lignin peroxidase in rhamnolipid(RL) reversed micelles

An environment friendly bio-surfactant of rhamnolipid(RL) was used as a solvent. The enzymatic reaction of oleic acid catalyzed by lipase and lignin peroxidase(lip) was evaluated. The optimum conditions of enzymatic reaction catalyzed by lipase(lip) were water to amphiphile molar ratio of 30(20), RL of 60(60) critical micelle concentration(CMC), pH of 7.0(3.0) and temperature of 40(30) °C, respectively. The change of enzyme conformation indicates that, for catalytic of lipase, water content is the most important factor of the enzymatic reaction of oleic acid, and p H for lip. With individual optimum conditions, the enzymatic efficiency of oleic acid catalyzed by lipase is higher than that by lip. In the presence of ethanol, the enzymatic reaction of oleic acid catalyzed by lipase suits Ping-Pong Bi-Bi mechanism. As an alternative to chemical reversed micelles, the RL reversed micelles are promising methods to enzymatic reaction of oleic acid.

Production and Recovery of Rhamnolipids Using Sugar Cane Molasses as Carbon Source

Biosurfactants were synthesized by Pseudomonas aeruginosa （P.A.）, using sugar cane molasses as carbon source. Assays were conducted in a shaker with agitation speed of 200 rpm, temperature of 38 ℃ and aeration ratio （Vm/Vf） of 0.4 and 0.6. A concentration of 3.0% was used for the carbon and energy source （molasses） and of 0.3% for the nitrogen source （NaNO3）. Samples were removed at regular times until 96 hours of cultivation. The reduction in surface tension was measured using the ring method; cell concentration was obtained by the dry mass and substrate consumption by the DNS method. The metabolite produced was extracted and quantified by the thioglycolic method. The results showed a maximum surface tension reduction of 46.57% after 60 h, 3.63 g/L of biomass after 8 h （μXmax =0.15 h^-1）, 79.60% of substrate consumption （μs= 0.67 h-1） and 4.47 g/L of rhamnolipid （μp=0.029 h^-1）.

Biosurfactant Production in Aerobic and Anaerobic Conditions by Different Species of the Genus Pseudomonas

In a time where surface active agents are capable of reducing the energy of the bonds between water molecules by interacting with them to reduce surface tension, it would be unwise not to be able to generate these in masses. Different Pseudomonas species were grown in MSP （minimal sulphate phosphate） media containing salts, glycerol and glucose. P. aeruginosa grown aerobically in the presence of glycerol as carbon source showed the highest emulsion percentage （81.48%）, most significant decrease in surface tension （20 mN/m） and rhamnose production of 2.86 mg/mL. However, in anaerobic conditions there was no emulsion, rhamnolipid production or decrease in surface tension. The rhamnolipids were molecularly characterized using ESI-MS （electrospray ionization-mass spectrometry）, P. aeruginosa CVCM 411 is able to produce mono-rhamnolipids and di-rhamnolipids, being rhamnolipid RhC10C12.1 the predominant monomer. The specific growth rate for isolates ofP. aeruginosa and P.fluorescens in MSP are 0.6732 ht and 0.2181 h^-1, respectively. In conclusion, the formation of rhamnolipids by P. aeruginosa is linked to its growth （depending on μ）, and its ability to generate about 35% of the μmax in the presence of glucose （carbon source） and glycerol （applied as pulses）.

Biotreatment of oily wastewater by rhamnolipids in aerated active sludge system

Oily wastewater generated by various industries creates a major ecological problem throughout the world. The tra- ditional methods for the oily wastewater treatment are inefficient and costly. Surfactants can promote the biodegradation of pe- troleum hydrocarbons by dispersing oil into aqueous environment. In the present study, we applied rhamnolipid-containing cell-free culture broth to enhance the biodegradation of crude oil and lubricating oil in a conventional aerobically-activated sludge system. At 20 ℃, rhamnolipids （11.2 mg/L） increased the removal efficiency of crude oil from 17.7% （in the absence of rham- nolipids） to 63%. At 25 ~C, the removal efficiency of crude oil was over 80% with the presence of rhamnolipids compared with 22.3% in the absence of rhamnolipids. Similarly, rhamnolipid treatment （22.5 mg/L） for 24 h at 20℃ significantly increased the removal rate of lubricating oil to 92% compared with 24% in the absence of rhamnolipids. The enhanced removal of hydrocarbons was mainly attributed to the improved solubility and the reduced interfacial tension by rhamnolipids. We conclude that a direct application of the crude rhamnolipid solution from cell culture is effective and economic in removing oily contaminants from wastewater.