石油降解菌XD-1产表面活性剂的性能

从含油废水中筛选分离到1株原油降解菌XD-1,鉴定为假单胞菌（Pseuomonas sp.）.初步实验表明菌XD-1具有较强的产表面活性剂乳化原油的作用,对该菌的产表面活性剂性能进行了研究.实验证明,菌XD-1所产表面活性剂为脂肽类物质,菌在生长对数期产表面活性剂,表面活性剂的产生为生长相关型;充足的碳源是产表面活性剂的必需条件,菌利用原油为碳源时能持续大量地产表面活性剂;原油和尿素为产表面活性剂的最适碳源和氮源,菌XD-1产表面活性剂的最佳营养培养基组成为葡萄糖10 g,尿素4 g,磷酸二氢钾1 g,微量元素液4 mL,水1 L,pH 8.0.     

海洋红球菌SY095产生物表面活性剂发酵培养基的优化

为提高海洋红球菌SY095发酵产生物表面活性剂的能力,研究发酵培养基中碳源及氮源的影响,并通过响应面分析法优化培养基各组分的含量。首先通过单因素实验,确定培养基最佳碳源、氮源分别为大豆油和尿素,在此基础上,利用Plackett-Burman实验设计筛选出2个对表面活性剂产量有显著影响的因子为尿素和K2HPO_4,通过最陡爬坡实验和响应面分析法对显著影响因子进行优化,获得海洋红球菌产表面活性剂的最佳培养基配方为:大豆油2%(体积分数),尿素2.35 g·L~(-1),K_2HPO_40.69 g·L~(-1),KH_2PO_41.0 g·L~(-1),Mg SO_40.5 g·L~(-1)。使用该优化培养基配方,海洋红球菌发酵液的表面张力为29.254 m N·m~(-1),与预测值接近,表面张力活性比优化前提高了12.9%。     

石油降解菌产表面活性剂的条件优化

为了了解陕北石油降解菌产表面活性剂的情况,对实验菌株CT-6以原油为唯一碳源时产表面活性剂的排油活性、表面张力和乳化性能进行了研究,并对实验菌株产表面活性剂的条件进行了优化.结果表明,排油圈直径达到8.0cm,表面张力降低到33 mN/m,乳化指数达89.9％,石油降解率达到68.0％;优化后的条件为:接种量10％、pH为8、温度28℃、转速220 r/min,该条件下,排油圈直径增加了13.1％,7d时乳化指数达到93.0％.     

一株生物表面活性剂产生菌的筛选鉴定及其特性

利用蓝色凝胶平板筛选法,从华北某油田受污染的土壤中分离筛选得到1株优良的生物表面活性剂产生菌H1.通过生理生化特征及16S rRNA基因序列相似性分析,将其鉴定为肺炎克雷伯氏菌(Klebsiella pneumoniae).薄层层析和红外光谱分析表明,该菌株所产生物表面活性剂为磷脂类生物表面活性剂.对其所产生物表面活性剂的稳定性进行研究,并考察影响该生物表面活性剂合成的因素.稳定性实验显示,该生物表面活性剂可耐受90℃的高温,对pH有较广泛的适应性(pH6.5～11.0),NaCl浓度对其生物活性影响不大.以蔗糖为碳源,硝酸铵为氮源,初始pH 6.5 ～7.0,30℃的培养条件有利于该生物表面活性剂的合成,生物表面活性剂的产量可达0.742 g/L.该研究成果可为原油的驱油降黏提供有效的菌源和理论依据.     

一株产表面活性剂石油降解菌筛选及其特性

为了从含有原油的样品中筛选出降解性能良好的产生表面活性剂菌株。利用富集、驯化培养、不同配方培养基筛选和排油活性测定,筛选出产高效生物表面活性剂的菌株。在形态特征和生理生化鉴定的基础上,对其16S rDNA序列进行了分析,并分别采用称重法和薄层层析法(TLC)测定其原油降解率和鉴别其表面活性剂的类型。进一步考察了不同种类的碳源和氮源、不同梯度的温度、盐度、酸碱度对其生长和排油活性的影响。筛选出一株高效脂肽类生物表面活性剂的菌株A-08,对其进行形态学、生理生化特性及16S rDNA序列同源分析,此菌株为荧光假单胞菌(Pseudomonas fluorescens)。A-08最适条件为碳源是食用油和液体石蜡,氮源为(NH_4)_2SO_4,温度40℃,盐度20%,pH 7.0,培养7 d后原油降解率为40.37%。实验筛选的菌株能适应较广泛的环境条件,可以对石油污染进行有效生物修复,具有较好的意义和应用价值。     

利用不同组分原油逐级驯化筛选高效石油烃降解混菌

利用不同组分原油逐级驯化的方法对克拉玛依油田的石油污染土样进行石油烃降解混菌的富集驯化,得到一组对稀油和稠油均具有高效降解能力的混菌M3。与采用单一原油驯化方法相比,混菌M3对稀油和稠油的降解率分别提高了12.5%和22%。该混菌具有较强的产表面活性剂的能力,能够使发酵液的表面张力从69.8 mN·m~(-1)降至27.9 mN·m~(-1)。通过混菌M3的生长条件优化实验得出:在温度30℃、pH 7～8、盐度1%、氮源选择尿素的条件下,混菌M3对原油的降解率最高。通过考察混菌M3在污染土壤中对原油的降解效果,发现:在修复期间,土壤脱氢酶呈先升高后降低的趋势;混菌M3可使饱和烃组分增加,并使芳香分、胶质和沥青质组分降低,对重质组分具有较好的降解效果。混菌M3的加入改变了原油性质,促进了土壤中原油的降解,经过56 d修复,土壤中原油降解率达到55.3%。     

产表面活性剂菌筛选及其对柴油降解影响研究

运用微生物培养方法,从饭店排污口污泥中筛选了7株产表面活性剂菌株。为了研究产表面活性剂菌在柴油降解过程中所产生的影响,分别将这些菌株与柴油降解菌(实验室提取的另一株假单胞菌)共同作用,其中一株(B_(26))不但可以将降解诱导期从6 d缩短至4 d,而且能将降解率由原来的71.1%提高至80.6%,具有较大的研究价值。实验着重对B_(26)的作用机理、柴油降解的影响因素等进行了分析研究。经形态学和生理生化实验鉴定,B_(26)属于芽孢菌属,其代谢过程中产生脂肽类表面活性剂,利用冷冻干燥法提取生物表面活性剂产量为1.57 g/L。     

影响原油生物降解因素的实验研究

以原油为唯一碳源,从长期被石油污染土壤中筛选出6株原油降解菌SY1~SY6,通过单因素实验研究初始pH、温度、充氧量(摇床转数)、盐浓度、氮源和磷源等环境因素和营养条件对各菌株生物降解原油的影响。实验结果表明:6株原油降解菌在初始pH7~9,温度30℃,摇床转速180 r/min时生长良好,且能有效地降解石油类污染物,其平均降解率为50%以上。6菌株在盐浓度在1%时生长良好,SY3菌和SY4菌能在盐浓度10%以上生长,具有一定的耐盐能力。同时,6菌株以氯化铵(NH4Cl)为氮源,磷酸氢二钾(K2HPO4)和磷酸二氢钾(KH2PO4)的混合物(2∶1)为磷源时生长良好,因此可作为各菌生长的最适氮源和磷源。研究结果可以为含油废水的处理提供微生物基础。     

培养基对菌产微生物絮凝剂的影响研究

研究了培养基成分种类及其用量对菌产微生物絮凝剂的影响。结果表明,较高的C/N比对菌产絮凝剂有利,碳源中的蔗糖是菌株Aeromonassp.N11产絮凝剂的良好碳源,氮源以采用复合氮源为佳。培养基中加入酵母膏有利于絮凝剂的产生。NaCl能够促进所产絮凝剂的絮凝活性,碳源蔗糖和氮源脲对菌产絮凝剂的影响最大。利用正交实验得出产絮凝剂培养基的最佳配比为蔗糖20g/L,酵母膏08g/L,脲05g/L,硫酸胺05g/L,NaCl7g/L。     

PVA降解混合菌系生长和产酶的营养条件分析

研究碳源、氮源以及不同金属离子对PVA降解混合菌系生长和产酶的影响。结果表明,混合菌系生长过程中无机氮源优于有机氮源;补充碳源可促进混合菌系的生长,但酶活比不添加时稍有降低。金属离子对酶活影响的正交实验表明Mg2＋浓度影响最大,其次是Fe2＋和K＋浓度,Ca2＋、Mn2＋、Zn2＋和Cu2＋浓度影响较小;最佳浓度分别为Mg2＋0.05 g/L、Fe2＋0.04 g/L、K＋2 g/L和Ca2＋0.05 g/L。     

Evaluation of biosurfactants for crude oil contaminated soil washing

An evaluation of the ability of aqueous biosurfactant solutions (aescin, lecithin, rhamnolipid, saponin and tannin) for possible applications in washing crude oil contaminated soil was carried out. The biosurfactants behaviour in soil-water, water-oil and oil-soil systems (such as foaming, solubilization, sorption to soil, emulsification, surface and interfacial tension) was measured and compared with a well-known chemical surfactant (sodium dodecyl sulphate, SDS) at varying concentrations. Results showed that the biosurfactants were able to remove significant amount of crude oil from the contaminated soil at different solution concentrations for instance rhamnolipid and SDS removed up to 80% oil and lecithin about 42%. The performance of water alone in crude oil removal was equally as good as those of the other biosurfactants. Oil removal was due to mobilization, caused by the reduction of surface and interfacial tensions. Solubilization and emulsification effects in oil removal were negligible due to the low crude oil solubilization of 0.11%. Therefore, these studies suggest that knowledge of surfactants' behaviour across different systems is paramount before their use in the practical application of oil removal.     

Improved bioavailability and biodegradation of a model polyaromatic hydrocarbon by a biosurfactant producing bacterium of marine origin

Polyaromatic hydrocarbons (PAHs) are organic pollutants mostly derived from the processing and combustion of fossil fuels and cause human health hazards. In the present study a marine biosurfactant producing strain of Bacillus circulans was used to increase the bioavailability and consequent degradation of a model polyaromatic hydrocarbon, anthracene. Although the organism could not utilize anthracene as the sole carbon source, it showed better growth and biosurfactant production in an anthracene supplemented glycerol mineral salts medium (AGlyMSM) compared to a normal glycerol mineral salts medium (GlyMSM). The biosurfactant product showed high degree of emulsification of various hydrocarbons. Analysis by gas chromatography (GC), high performance thin layer chromatography (HPTLC) and Fourier transform infrared spectroscopy (FTIR) showed that the biosurfactant could effectively entrap and solubilize PAH. Thin layer chromatographic analysis showed that anthracene was utilized as a carbon substrate for the production of biosurfactant. Thus organic pollutant anthracene was metabolized and converted to biosurfactants facilitating its own bioremediation.     

Factors affecting biosurfactant production by oil degrading Aeromonas spp. isolated from a tropical environment

An Aeromonas spp. was isolated from tropical estuarine water. The organism grew on crude oil and produced biosurfactant that could emulsify hydrocarbons. The peak growth and biosurfactant production was on the 8th day. The organism grew on a range of hydrocarbons that include crude oil and hexadecane while no growth was recorded on some hydrocarbons that include benzene. The biosurfactant produced by the organism emulsified a range of hydrocarbons with diesel (E24=65) as the best substrate and hexane (E24=22) as the poorest. After purification, the biosurfactant was found to contain about 38% carbohydrate and an unidentified lipid. No protein was present in the purified biosurfactant. Production of biosurfactant was highest in medium with glucose and lowest in the medium with diesel+acetate. Soybean was the best nitrogen source for biosurfactant production. The activity of the biosurfactant was enhanced optimally at NaCl concentration of 5%, pH of 8.0 and temperature of 40 degrees C. The biosurfactant retained 77% of its original activity after 120 min of exposure to heat at a temperature of 100 degrees C. Biosurfactant may be produced with this organism using non-hydrocarbon substrates such as glucose and soybean that are readily available and would not require extensive purification for use in food and pharmaceutical industries.     

Characterization of hydrocarbon-degrading and biosurfactant-producing Pseudomonas sp. P-1 strain as a potential tool for bioremediation of petroleum-contaminated soil

The Pseudomonas sp. P-1 strain, isolated from heavily petroleum hydrocarbon-contaminated soil, was investigated for its capability to degrade hydrocarbons and produce a biosurfactant. The strain degraded crude oil, fractions A5 and P3 of crude oil, and hexadecane (27, 39, 27 and 13 % of hydrocarbons added to culture medium were degraded, respectively) but had no ability to degrade phenanthrene. Additionally, the presence of gene-encoding enzymes responsible for the degradation of alkanes and naphthalene in the genome of the P-1 strain was reported. Positive results of blood agar and methylene blue agar tests, as well as the presence of gene rhl, involved in the biosynthesis of rhamnolipid, confirmed the ability of P-1 for synthesis of glycolipid biosurfactant. ¹H and ¹³C nuclear magnetic resonance, Fourier transform infrared spectrum and mass spectrum analyses indicated that the extracted biosurfactant was affiliated with rhamnolipid. The results of this study indicate that the P-1 and/or biosurfactant produced by this strain have the potential to be used in bioremediation of hydrocarbon-contaminated soils.     

The production of rhamnolipid by a Pseudomonas aeruginosa strain isolated from a southern coastal zone in Brazil

The production and properties of a rhamnolipid-type biosurfactant, synthesized by the Pseudomonas aeruginosa LBM10 strain, isolated from a southern coastal zone in Brazil, were investigated. The assays were conducted in a rotary shaker at 30 degrees C and 180 rpm for a period of 96 h. Soybean oil and sodium nitrate were the best sources of carbon and nitrogen, respectively. A nitrogen-limiting condition (C/N ratio of 100) was favorable to biosurfactant production. The formation of stable emulsions was better in saline concentrations below 0.5%, pH values in the range from 6 to 9 and temperatures in the range from 35 to 40 degrees C, maintaining about 80% of its original activity for salinity up to 3% and 120 min of exposure at 100 degrees C. The biosurfactant may be produced with this microorganism using renewable substrates that are readily available, reaching values of 1.42 g l(-1) measured as rhamnose. This biosurfactant has interesting and useful properties for many industrial applications.     

Isolation and characterization of a potential paraffin-wax degrading thermophilic bacterial strain Geobacillus kaustophilus TERI NSM for application in oil wells with paraffin deposition problems

Paraffin deposition problems, that have plagued the oil industry, are currently remediated by mechanical and chemical means. However, since these methods are problematic, a microbiological approach has been considered. The bacteria, required for the mitigation of paraffin deposition problems, should be able to survive the high temperatures of oil wells and degrade the paraffins under low oxygen and nutrient conditions while sparing the low carbon chain paraffins. In this study, a thermophilic paraffinic wax degrading bacterial strain was isolated from a soil sample contaminated with paraffinic crude oil. The selected strain, Geobacillus TERI NSM, could degrade 600mg of paraffinic wax as the sole carbon source in 1000ml minimal salts medium in 7d at 55 degrees C. This strain was identified as Geobacillus kaustophilus by fatty acid methyl esters analysis and 16S rRNA full gene sequencing. G. kaustophilus TERI NSM showed 97% degradation of eicosane, 85% degradation of pentacosane and 77% degradation of triacontane in 10d when used as the carbon source. The strain TERI NSM could also degrade the paraffins of crude oil collected from oil wells that had a history of paraffin deposition problems.     

高效石油烃降解菌SKD-1的分离、筛选及其降解性能

从孤岛油田石油污染土壤中分离到一株高效石油降解菌,命名为SKD-1。该菌株菌落表面湿润光滑、边缘整齐、圆形、不透明、乳黄色,能够利用葡萄糖和淀粉作为其生长的碳源和能源,其最适生长环境为碱性（pH8-10）,在分别以正十六烷烃和原油为惟一碳源,温度为30℃,摇床（180r／min）培养的条件下,菌株SKD-1的降解率分别为66．1％和36．9％。16SrRNA基因序列分析表明,菌株SKD-1与不动杆菌AcinetobactercalcoaceticusSY-1同源性达99％。结合菌株SKD．1菌落形态、理化性质以及系统发育分析,可以鉴定菌株SKD-1属于不动杆菌属（Acinetobactersp．）,序列登录号为AB774229。     

炼厂"三泥"的生物处理研究

从大港油田筛选和分离出3株以原油为碳源的石油降解菌,初步鉴定为假单胞菌属(Pseudomonas sp.),用这3株菌混合处理某炼厂的"三泥",在适宜的降解条件下,仅需投加混合菌液,无需添加营养物(N,P),可使含油污泥A的残油量从67g/kg降至7.7g/kg,混合油泥试样AB的残油量从19.7g/kg降至3.1g/kg,平均去除率为84%以上,可以实现含油污泥的较为经济的、有效的处理.经生物处理后含油污泥的恶臭味完全消失.     

Isolation, identification and characterization of Bacillus amyloliquefaciens BZ-6, a bacterial isolate for enhancing oil recovery from oily sludge

Over 100 biosurfactant-producing microorganisms were isolated from oily sludge and petroleum-contaminated soil from Shengli oil field in north China. Sixteen of the bacterial isolates produced biosurfactants and reduced the surface tension of the growth medium from 71 to <30 mN m⁻¹ after 72 h of growth. These bacteria were used to treat oily sludge and the recovery efficiencies of oil from oily sludge were determined. The oil recovery efficiencies of different isolates ranged from 39% to 88%. Bacterial isolate BZ-6 was found to be the most efficient strain and the three phases (oil, water and sediment) were separated automatically after the sludge was treated with the culture medium of BZ-6. Based on morphological, physiological characteristics and molecular identification, isolate BZ-6 was identified as Bacillus amyloliquefaciens. The biosurfactant produced by isolate BZ-6 was purified and analyzed by high performance liquid chromatography-electrospray ionization tandem mass spectrometry. There were four ion peaks representing four different fengycin A homologues.