Biosurfactant production by <Emphasis Type="Italic">Bacillus coagulans</Emphasis>

A new biosurfactant producer, Bacillus coagulans, was isolated from soil. Its 24-h-old culture broth had a low surface tension (27–29 mN/m). Optimization of cell growth of this bacterium led to maximal biosurfactant production with glucose or starch as the organic carbon source, a pH in the range 4.0–7.5, and incubation temperatures from 20 to 45°C. The crude biosurfactants obtained after neutralization and lyophilization of the acid precipitate yielded a minimal aqueous solution surface tension value of 29 mN/m and an interfacial tension value of 4.5 mN/m against hexadecane. The critical micelle concentration of the crude biosurfactants was 17 mg/L. Addition of NaCl to the aqueous solution of the crude product caused lowering of surface tension at both the aqueous solution-air and aqueous solution-n-hexadecane interfaces. These results indicate that the biosurfactants obtained have potential environmental and industrial applications and may have uses in microbially enhanced oil recovery.     

一株脂肽类生物表面活性剂产生菌的性能及其影响因素研究

通过对实验筛选出一株产脂肽类生物表面活性剂的菌株BW-23,进行性能及其影响因素研究.结果表明:脂肽表面活性剂的CMC浓度为70 mg/L时,表面张力为31.0 mN/m;发酵液稀释10倍后,乳化指数仍为24.72％;在10～60℃间菌株BW-23所产的生物表面活性剂的耐热性较好;菌株BW-23在pH值为5.0～8.0时表面活性最好;当NaCl浓度小于15％时,菌株BW-23水溶液的表面张力变化幅度最小;当Ca2+浓度达到5000 mg/L时,菌株BW-23水溶液的表面张力为30.5 mN/m.     

Sunflower Acid Oil-Based Production of Rhamnolipid Using Pseudomonas aeruginosa and Its Application in Liquid Detergents

Utilization of industrial waste as substrates for the rhamnolipid synthesis by Pseudomonas aeruginosa is a worthy alternative for conventionally used vegetable oils and fatty acids to reduce the production cost of rhamnolipid. Sunflower acid oil (SAO), a by-product of the oil industry, contains 70% 18:0 fatty acid, with oleic acid as a major component. In this scope, production and analysis of rhamnolipid was successfully demonstrated using SAO as a new substrate. Pseudomonas aeruginosa produced rhamnolipid (a glycolipid biosurfactant) at a maximum concentration of 4.9 g L⁻¹ with 60 g L⁻¹ of SAO in the medium. Structural properties of rhamnolipid biosurfactant are confirmed using thin layer chromatography (TLC), high performance liquid chromatography (HPLC), and fourier transformed infrared spectroscopy (FTIR) analysis. Further surface-active properties of the crude rhamnolipid were evaluated by measuring surface tension and emulsification properties. The synthesized rhamnolipid reduced the surface tension of water to 30.12 mN m⁻¹ and interfacial tension (against heptane) to 0.52 mN m⁻¹. Moreover, rhamnolipid shows the highest emulsification index (above 80%) for vegetable oils. This study confirms the use of SAO as a potential substrate for rhamnolipid production. The synthesized rhamnolipid was incorporated in liquid detergent formulation along with alpha olefin sulfonate (AOS) and sodium lauryl ether sulfate (SLES). The performance properties including foaming and cleaning efficiency of liquid detergent were compared.     

脂肽-糖脂混合型生物表面活性剂的性能及助剂

分离提取了枯草芽孢杆菌Bacillus subtilis THY-8发酵产物中的生物表面活性剂,采用飞行时间质谱鉴定为鼠李糖脂和表面活性素、芬芥素等的混合物。考察了该混合型生物表面活性剂的性能,研究了其对油砂原油的驱油效率,并探索了合适的助剂种类及配伍浓度。结果表明,THY-8所产的脂肽-糖脂混合型生物表面活性剂具有良好的表面活性、热稳定性和乳化性,pH值为5.5时表面张力为27.59 mN/m,临界胶束浓度为15 mg/L;在70 ℃放置5天,表面张力基本不变;可将液体石蜡乳化形成粒径10～30 μm的乳液。正辛醇助剂与该混合生物表面活性剂复配后界面张力降低到10?3 mN/m,驱油效率提高3.2倍。含0.9 g/L生物表面活性剂的发酵液驱油效果与正辛醇-生物表面活性剂复配体系相当。     

Optimization and Characterization of Biosurfactant Rhamnolipid Production by Pseudomonas aeruginosa Isolated from an Artificially Contaminated Soil

Biosurfactants are surfactants biologically produced by microorganisms, presenting several advantages when compared to synthetic surfactants. Pseudomonas aeruginosa is known for producing rhamnolipids, considered one of the most interesting types of biosurfactants due to their high yields, when compared to other types. In this work, the production of rhamnolipid from P. aeruginosa was optimized. At first, the Plackett–Burman design was used to select most significant variables affecting the biosurfactant production yield among nine variables—carbon–nitrogen ratio, carbon concentration, nitrogen source, pH, cultivation time, potassium and magnesium concentrations, agitation, and temperature. Then, using main variables, a central point experimental design aiming to optimize rhamnolipid production was performed. The maximum biosurfactant concentration obtained was 0.877 mg L⁻¹. The rhamnolipid also displayed a great emulsification rate, reaching approximately 67%, and the ability to reduce water surface tension from 72.02 to 35.26 mN m⁻¹ at a critical micelle concentration (CMC) of 127 mg L⁻¹, in addition to presenting a good stability when exposed to wide pH and salinity ranges. The results suggest that rhamnolipids are promising substitutes for synthetic surfactants, especially due to lower impacts on the environment.     

Recycling of Bakery Waste as an Alternative Carbon Source for Rhamnolipid Biosurfactant Production

Production of a rhamnolipid biosurfactant (RBS) using discarded mixed bakery waste (BW) employing bacterial strain Pseudomonas aeruginosa strain PG1 (identified by 16 s rDNA sequencing) was investigated for bioconversion of the food waste. Dry and powder form BW was supplemented with mineral salt media (MSM) as a sole carbon source for production of RBS. RBS production was measured based on the drop collapse assay and surface tension (ST) reduction of the culture media. Production of RBS in the culture media was enhanced by optimizing the carbon source (BW) concentration and the proper nitrogen source along with the pH of the MSM. Under optimized culture conditions, 11.56 g L⁻¹ day⁻¹ crude biosurfactant (BS) was achieved. The RBS had the ability to reduce the ST of the optimized MSM from 72.0 to 25.8 mN m⁻¹ during culture, where the critical micelle concentration (CMC) of the biosurfactant was found to be 100 mg L⁻¹. Liquid Chromatography Mass Spectroscopy (LC-MS), Fourier Transform Infrared spectroscopy (FTIR), and scanning electron microscopy (SEM)–energy dispersive X-ray spectroscopy (EDS) analyses of the purified BS confirmed that it is of rhamnolipid in nature and it is made up of both monorhamnolipid and dirhamnolipid congeners. Furthermore, the RBS did not express any cytotoxic effect on the cell line of mouse L292 fibroblastic cell indicating the biosafety nature of the high-value biomolecule.     

假单胞菌O-2-2产鼠李糖脂的结构表征及理化性质

利用液相色谱/质谱联用仪分析了铜绿假单胞菌O-2-2以正十八烷为碳源所产鼠李糖脂生物表面活性剂的组成。共检出21种鼠李糖脂的同系物,都由1~2分子的鼠李糖和1~2个含β-羟基的碳链长度为8~12的饱和或不饱和脂肪酸构成;主要组分为α-L-吡喃鼠李糖苷-β-羟基癸酰-β-羟基癸酸和2-O-α-L-吡喃鼠李糖苷-α-L-吡喃鼠李糖苷-β-羟基癸酰-β-羟基癸酸。该糖脂类生物表面活性剂可将水的表面张力降至28·6mN/m,临界胶束浓度为1·3×10-4mol/L,在120℃加热4h或者在ρ(NaCl)=100g/L或ρ(CaCl2)=20g/L的盐溶液中仍能保持表面活性。     

Use of biosurfactant in the removal of oil from contaminated sandy soil

The effectiveness of cell‐free rhamnolipid biosurfactant, derived from the culture medium at the end of fermentation was investigated for the removal of two different kinds of oil from contaminated sandy soils. The crude cultivation medium, containing 13.2 g L^?1 of rhamnolipids, had a surface tension, interfacial tension and critical micellar concentration of 30 mN m^?1, 2 mN m^?1 and 60 mg L^?1, respectively. The evaluation of biosurfactant in the culture medium (BM) and oil concentrations in the removal of oil from different contaminated sandy soil was performed using a statistical experimental design tool. Oil in sandy soil, containing predominantly aromatic or paraffinic hydrocarbons (5 to 10% w/w), was removed by as much as 91 and 78%, respectively, in the presence of reduced amounts of BM (6.3 to 7.9 g L^?1). The progress of oil removal was monitored for 101 days and results indicated that removal efficiency in sandy soil with aromatic characteristics was relatively stable over the entire period. Based on these studies, it is concluded that use of a BM was effective in reducing oil concentrations in contaminated sandy soil. Copyright © 2007 Society of Chemical Industry     

E. coli O157:H7 Desorption by Rhamnolipid Biosurfactant in Water-Unsaturated Porous Media

Animal waste is a valuable resource, which can add organic matter, nitrogen, phosphorous and potassium as well as other nutrients needed for plant growth when it is used in the agricultural field. When applied to an agricultural field, infectious agents or pathogenic organisms in the animal waste are usually retained in the soil. At the same time, surfactants are popularly utilized in pesticide applications to aid pesticide solubility and mobility. Consequently, the retained pathogenic strains might be flushed out of the soil matrices with a possibility of contaminating the groundwater. In this research, we investigated desorption of E. coli O157:H7 pre-deposited in silica sand and its subsequent release and transport in the presence of rhamnolipid biosurfactant in unsaturated laboratory columns. Based on the experimental observations, it was discovered that retained E. coli O157:H7 was released during flushing with rhamnolipid biosurfactant solutions. E. coli O157:H7 release rate coefficient increased both with the increase of rhamnolipid biosurfactant concentration and the increase of water saturation. With the increase of rhamnolipid biosurfactant concentration, the air–water surface tension decreased. With the increase of water saturation, the air–water interfacial area decreased. Both the decreased air–water surface tension and air–water interfacial area favored E. coli O157:H7 release. Increase of E. coli O157:H7 release rate coefficient with the decrease of air–water surface tension was more pronounced for low air–water interfacial area (i.e., high water saturation). Similarly, increase of E. coli O157:H7 release rate coefficient with the decrease of air–water interfacial area was more pronounced for low air–water surface tension (i.e., high rhamnolipid biosurfactant concentration). E. coli O157:H7 retention in unsaturated systems was found to be controlled by the capillary force, which was greatly impacted by water saturation and air–water surface tension. Increase of water saturation and rhamnolipid biosurfactant concentration both contributed to the decrease of the capillary force, resulting in increased E. coli O157:H7 release rate.     

生物破乳菌的培养及代谢产物分析

从胜利油田采出水及其它样品中筛选得到破乳效果较好的D3破乳菌,考察了该破乳菌的培养条件及代谢产物。结果表明,D3菌的适宜生长条件为:温度40℃,pH 7~9,培养时间72 h。D3菌为革兰氏阴性细菌,归为沙雷氏菌属;其代谢产物主要含有:鼠李糖酯、脂肽类、醛、胺。D3菌质量浓度10 g/L时,油水的界面张力由7.65 mN/m降至0.98 mN/m;当菌浓度在0.5~1.0 g/L时,含油污水的除油率大于50%。     

无患子皂素水提液发酵精制联产表面活性剂鼠李糖脂

以无患子皂素水提水解液为底物,发酵生产生物表面活性剂,在精制无患子皂素的同时发酵联产生物表面活性剂鼠李糖脂。经铜绿假单胞菌发酵后发酵完成液中不含葡萄糖,葡萄糖的消耗速率与接种量成正相关,在发酵液中额外添加大豆油可促进表面活性剂亲油基团的生成,溶液表面活性进一步提高。发酵后溶液中表面活性物质浓度达到50.8g/L,比对照组提高了16.1%,溶液表面张力明显降低。接种10%的菌种发酵获得的无患子皂素复合产物干粉其临界胶束浓度由10g/L降低到2.5g/L,临界胶束浓度下的复合产物水溶液表面张力比未接种菌种的低18.67%,复合产物具有很好的起泡性能及更高的泡沫稳定性。     

不同腰果酚磺酸盐的合成及界面活性

以天然生物腰果酚为原料,经过磺化和中和两步反应合成了两种腰果酚磺酸盐表面活性剂.FT-IR光谱证实两种腰果酚磺酸盐的化学结构.采用滴体积表面张力仪和全自动旋滴界面张力仪测定了两种腰果酚磺酸盐水溶液的表面张力和油水界面张力,结果表明,两种生物质腰果酚磺酸盐具有良好的表面活性和界面活性,25℃时侧链不饱和的腰果酚磺酸盐和侧链饱和的腰果酚磺酸盐的临界胶束浓度分别为38.1mg/L和28.2mg/L,此浓度下的表面张力分别38.54mN/m和37.35mN/m;饱和腰果酚磺酸盐质量分数高于0.8%时,可将油水界面张力降低至10^-3mN/m,但不饱和腰果酚磺酸盐仅能将油水界面张力降低至10^-1mN/m.侧链饱和的腰果酚磺酸盐的表面活性和界面活性均优于侧链不饱和的腰果酚磺酸盐.     

N-丙基-N-全氟辛基磺酰基甘氨酸盐类表面活性剂的合成方法改进及性能

以国产全氟辛基磺酰氟为原料 ,与正丙胺在异丙醚中 5 0℃下反应制得到N 丙基全氟辛基磺酰胺 ,再与氯乙酸乙酯在碱性条件下回流缩合得N 丙基 N 全氟辛基磺酰基甘氨酸乙酯 ,水解后得到N 丙基 N 全氟辛基磺酰基甘氨酸 ,总产率 79%。其锂、钠、钾盐的CMC值分别是 6 72mg/L、2 1 66mg/L及 3 0 79mg/L ,在质量浓度为 80mg/L时表面张力均降至 2 0mN/m以下 ,说明该类产品具有良好的表面活性     

菌株Pseudomonas aeruginosa BC1发酵-渗透汽化耦合制备鼠李糖脂的研究

采用发酵-渗透汽化耦合(fermentation coupling with pervaporation,FCP)系统和分批补料式发酵系统培养Pseudomonas aeruginosa BC1制备生物表面活性剂鼠李糖脂,两轮发酵过程分别持续124 h和107 h。在FCP系统中,最大细胞吸光度OD600为1.06;生物表面活性剂OD600为0.61;鼠李糖脂最终产量为1.3 g/L,相比于分批补料式发酵系统提高38%;对比纯水的表面张力67.52 m N/m,发酵第29 h的发酵上清液表面张力为22.56 m N/m。同时,该菌的发酵上清液对液体石蜡、机油、柴油、正己烷、十六烷均有较好的乳化能力。经GC-MS结合SPME分析发现,FCP系统分离出的渗透蒸汽冷凝液中含有乙醇、戊醇等有机物。实验结果表明,相比分批补料式发酵系统,FCP系统能够分离发酵过程中产生的一部分挥发性代谢产物,减轻这些物质对细胞生长的抑制,使细胞浓度和鼠李糖脂产量都有明显提高。     

Biodegradation of Petroleum Oil Effluents and Production of Biosurfactants: Effect of Initial Oil Concentration

Biosurfactants have been used in the petroleum oil industry for oil storage tank cleaning, for reducing the viscosity of heavy oil, thereby facilitating recovery, transportation, and pipelining. Also, they are used for microbial enhanced oil recovery either from residual oil in reservoirs or from oily wastewater. The present study is an investigation of biodegradation of petroleum oil effluents using Pseudomonas aeruginosa ATCC 9027 for producing rhamnolipid biosurfactants. The processes were performed in a mechanically agitated, fully baffled, air-sparged 10 L glass fermenter with a 5 L working volume. The effect of oil concentration (1%, 1.5%, 2%, and 2.5%) on the efficiency of oil biodegradation, rhamnolipid production, surface tension, and bacterial biomass was studied. The fermentation with 1.5% oil concentration gave the highest biodegradation for aliphatic hydrocarbons (98.85%), followed by 2% oil concentration (95% degradation), then with 2.5% oil concentration (80% degradation), and finally with 1% oil concentration (66.8% degradation). Also, the complete biodegradation of polyaromatic hydrocarbons was achieved for all tested oil concentrations. On the other hand, maximum rhamnolipid production of 2.7 g L⁻¹ as rhamnose equivalent and lower surface tension (30.2 mN m⁻¹) were achieved at 2% oil concentration.     

沥青质模型油/MD膜驱剂溶液的界面张力

从MD膜驱剂与原油界面活性组分沥青质模型油的界面张力出发,考察了作用时间、水相pH、MD膜驱剂质量浓度、盐浓度、沥青质含量、芳香度、温度等对模型油/水界面张力的影响,并根据扩散控制机理解释了动态界面张力初期过程,进一步揭示了MD膜驱油技术的机理。结果表明,沥青质模型油/MD膜驱剂溶液的界面张力先随时间增加而降低,约15min后达到平衡,达到平衡之前的过程基本上符合扩散控制过程;MD膜驱剂的加入并不能改变pH对模型油/水界面张力的影响趋势;在所考察的条件范围内,沥青质模型油/水溶液的界面张力不随MD膜驱剂质量浓度增加而改变,其值约为19 65mN/m;NaCl对界面张力的影响不明显;沥青质质量浓度从0增加到1000mg/L时,模型油/水和模型油/MD膜驱剂溶液的界面张力分别从23 4mN/m和22 0mN/m逐渐下降至20 0mN/m和18 8mN/m;温度从25℃升高到45℃时,模型油/水溶液的界面张力降低;但芳香度从0增加到100%时,其界面张力均从21 0mN/m增加至31 5mN/m。MD膜驱剂是表面非活性物质,在驱油时不存在低界面张力提高采收率的机理。     

表面活性剂的复配及对甲维盐微乳剂物理稳定性的影响

通过测定不同类型的单一及复配型表面活性剂水溶液的临界胶束浓度和表面张力,研究了它对w(甲维盐)=1％的微乳剂物理稳定性的影响。膦酸酯类阴离子表面活性剂A的临界胶束浓度为1．79×10-4 mol／L,表面张力为28．90 mN/m;苄基酚聚氧乙烯聚氧丙烯醚嵌段型非离子表面活性剂B(EPE型)的临界胶束浓度为 1.91×10-4 mol/L,表面张力为20．70 mN/m;按m(A):m(B)=2:3形成的复配型表面活性剂2#的水溶液的临界胶束浓度为9．30×10-5 moL/L,表面张力为25．66 mN/m。当w(2#)=10％时,配制w(甲维盐)=1％的微乳剂物理稳定性最佳,各项指标均合格。     

Biosurfactant from <Emphasis Type="Italic">Lysinibacillus chungkukjangi</Emphasis> from Rice Bran Oil Sludge and Potential Applications

A newly discovered bacterium, Lysinibacillus chungkukjangi, was isolated from the sludge of rice bran oil processing. When the bacterium was grown on rice bran, it produced biosurfactant which reduced the surface tension of the media to 27.9 from 72 mN/m. The biosurfactant was recovered by a solvent extraction method and characterized with the help of various structure elucidation techniques viz. FTIR, ¹H- and ¹³C-NMR spectroscopy and LC–MS analysis. The combined results of FTIR and NMR revealed the presence of carbonyl, olefinic and aliphatic groups, with the typical spectra of lipids. Moreover, LC–MS analysis also supported the same information. The biosurfactant was also studied for its anti-oxidant and microbial enhanced oil recovery (MEOR) potential. The anti-oxidant activity was observed by the DPPH free radical scavenging method using ascorbic acid as the standard. The IC₅₀ (the half maximal inhibitory concentration) was calculated and for standard, it was 0.056 mg/mL and for the biosurfactant it turned out to be 1.3 mg/mL which shows its good anti-oxidant potential. The sandpack test was performed to check its MEOR potential and kerosene was recovered up to 90 %, which shows its excellent applicability in the MEOR processes.     

Production and stability studies of the biosurfactant isolated from marine Nocardiopsis sp. B4

A potential biosurfactant producing strain, marine Nocardiopsis B4 was isolated from the West coast of India. Culture conditions involving variations in carbon and nitrogen sources were examined at constant pH, temperature and revolutions per min (rpm), with the aim of increasing productivity in the process. The biosurfactant production was followed by measuring the surface tension, emulsification assay and emulsifying index E24. Enhanced biosurfactant production was carried out using olive oil as the carbon source and phenyl alanine as the nitrogen source. The maximum production of the biosurfactant by Nocardiopsis occurred at a C/N ratio of 2:1 and the optimized bioprocess condition was pH 7.0, temperature 30° C and salt concentration 3%. The production of the biosurfactant was growth dependent. The surface tension was reduced up to 29 mN/m as well as the emulsification index E24 was 80% in 6 to 9 days. Properties of the biosurfactant that was separated by acid precipitation were investigated. The biosurfactant activity was stable at high temperature, a wide range of pH and salt concentrations thus, indicating its application in bioremediation, food, pharmaceutical and cosmetics industries.     

一种含氟烷基磷酸酯类氟碳表面活性剂的复配研究

研究了含氟烷基磷酸单酯类表面活性剂[分子式为H(CF2)6CH2OPO(ONa)2,记为DFH-PS]与无机盐和普通碳氢表面活性剂的复配性能,研究结果表明,DFH-PS水溶液最低表面张力为23.73 mN/m;当NaCl浓度为0.2 mol/L时,可使DFH-PS水溶液最低表面张力下降到21.62 mN/m;阴离子碳氢表面活性剂十二烷基硫酸钠(SDS)对该含氟表面活性剂影响显著,当n(DFH-PS)∶n(SDS)=5∶1时,可使水溶液表面张力在很低浓度时降至22.22 mN/m;与非离子表面活性剂辛基酚聚氧乙烯(10)醚(OP-10)混合,当n(DFH-PS)∶n(OP-10)=8∶1时,可使水溶液表面张力降至27.0 mN/m。