一株对硝基苯胺降解菌Microbacterium sp. PNA8的分离鉴定及其降解条件研究

从污泥中分离得到一株能以对硝基苯胺为唯一碳源、氮源和能源生长的细菌菌株PNA8。经过对其形态特征、生理生化特性、以及16S rRNA序列分析, 该菌株初步鉴定为Microbacterium sp.。进一步研究表明, 菌株PNA8利用对硝基苯胺生长和降解的最适温度和pH分别是30°C和7.0。培养基中添加定量酵母膏有利于菌株的生长及其对对硝基苯胺的降解。最适条件下, 在培养液中添加0.4 g/L酵母膏, 4 d内0.3 mmol/L对硝基苯胺降解率可达100%。     

腾冲热海七株高温厌氧菌的分离及鉴定

摘要：【目的】了解中国云南腾冲热海环境中厌氧菌的分类学特征及生理生化特性。【方法】利用Hungate厌氧操作技术,从云南腾冲热海80-93℃温泉中分离出7株高温厌氧菌,对其进行形态、生长特征及16S rRNA基因序列分析,确定菌株的系统发育地位。【结果】7株菌株细胞形态均为杆状,不产芽孢,革兰氏阴性,严格厌氧,在70℃生长良好。其中较典型的菌株RH0802能在55-80℃温度范围内生长,最适生长温度为70℃;生长pH值范围为5.5-8.5,最适pH值为7.0,能利用葡萄糖、淀粉、甘露醇、甘露糖、核糖、麦芽糖、纤维二糖、木糖、果糖、半乳糖、木聚糖、甘油,不能利用蔗糖、丙酮酸。16S rRNA基因序列相似性的比较分析表明,其中5株菌与Caldanaerobacter属菌株的最高相似性均在98%以上,而RH0804与RH0806分别为96%和93%。菌株RH0802-RH0808的序列登录号分别为FJ748766、FJ748762、FJ748761、FJ748763、FJ748765、FJ748764和FJ748767,在中国普通微生物菌种保藏中心的保藏号分别为CGMCC1.5134-1.5140。【结论】分离自腾冲热海的7株嗜热厌氧菌与Caldanaerobacter属菌株有较高相似性,可将这7株菌株鉴定为Caldanaerobacter属菌株（Caldanaerobacter. sp）,其中菌株RH0804和RH0806有成为新种的可能。     

一株解磷中度嗜盐菌的分离鉴定及解磷特性分析

从四川自贡某盐井壁植物根系土壤中分离得到1株中度嗜盐解磷菌QW1011。该菌细胞呈线状, 大小为0.8 μm×30 μm~100 μm, 革兰氏染色为阳性、最适NaCl生长浓度为10%, NaCl最高耐受浓度15%。好氧生长, 酪素水解、硝酸还原和接触酶阴性。菌株的16S rRNA基因序列(接受号：EF647207)与Bacillus megatherium ATCC 14581的16S rRNA相似性为100%, 其16S-23S rRNA间区(ISR)的PCR扩增片的PAGE指纹图谱与参考菌株Baci     

腾冲热海一株嗜酸热硫化叶菌的分离与鉴定

从云南腾冲热海酸性温泉中分离纯化出一株极端嗜酸热菌株K4-1,并对其进行形态观察、生长特征、碳源和能源利用及16S rRNA基因分析.该菌株细胞形态为不规则球形,有单生鞭毛,严格好氧,兼性自养,能利用元素硫作为能源,也能利用酵母膏、精氨酸或核糖作为碳源和能源.其最适生长温度为75℃,最适pH为3.5.通过16S rRNA基因序列相似性对比对该菌株进行鉴定,结果表明该菌株与硫化叶茵属标准菌株的相似性介于86.6%～94.3%之间,与分离自腾冲热海的腾冲硫化叶菌Sulfolobus tengchongensis RT8-4菌株序列相似性最高,达到98.9%,可将菌株K4-1鉴定为硫化叶菌属菌株.菌株K4-1的16S rRNA基因序列号为EU729124.     

一株石油烃降解菌新种Marinobacter sp. PY97S的鉴定

【目的】为了对1株从黄海沉积物中分离到的石油烃降解菌新种PY97S进行分类学鉴定。【方法】采用16S rRNA基因序列同源性分析、生理生化指标测定、抗生素抗性实验、DNA G+C含量测定、全细胞脂肪酸组成测定、碳源利用实验、呼吸醌测定以及DNA杂交实验等多种方法对该菌株进行鉴定,并通过降解实验测定其对烷烃的利用情况。【结果】菌株PY97S为海杆菌(Marinobacter),革兰氏阴性,接触酶阳性,氧化酶阳性,主要呼吸醌为Q-9。在GenBank中与其16S rRNA基因序列相似度最高的模式株为Marinobacter koreensisDD-M3T(96.93%),两者DNA-DNA同源性仅为46.7%。菌株PY97S的温度生长范围为15℃-35℃(最适为30℃),NaCl浓度生长范围是0%-10%(最适为0%),初始pH生长范围为pH 6.0-9.0(最适为初始pH7.0)。该菌株可以利用多种糖类和有机酸类的碳源,并对氨苄青霉素、氧哌嗪青霉素等多种抗生素敏感。其DNA G+C含量为48.2 mol%。其主要脂肪酸组成为2-methyl C15∶0(29.97%)、C16∶1ω7c(27.22%)、C12∶0(22.22%)和C16∶1ω9c(5.73%)。【结论】菌株PY97S是1株能够降解多种多环芳烃和烷烃的海洋石油烃降解菌新种,具有应用到溢油污染海洋环境生物修复的潜力。     

制革废水中壬基酚聚氧乙烯醚降解菌的分离及特性

从长期受壬基酚聚氧乙烯醚污染的制革废水中采集水样,通过富集培养的方法从中筛选到一株可以壬基酚聚氧乙烯醚为唯一碳源生长的降解菌OPQa3。通过形态学特征观察和生理生化试验,结合16S rRNA基因序列分析,初步鉴定菌株OPQa3属于短波单胞菌属Brevundimonas sp.,其16S rRNA基因序列与Brevundimonas diminuta(EU434566.1)的相似性最高(99%)。降解菌OPQa3的生长周期为24 h;以2%的量接种菌株OPQa3,保证最终OD600=0.70,于746 mg/L的壬基酚聚氧乙烯醚培养基中,120 h内的降解率可达84.5%,菌OPQa3生长的最佳温度为30°C,最适pH值在7左右。降解性质粒检测结果表明,控制菌株OPQa3降解壬基酚聚氧乙烯醚的基因位于质粒上。     

同步脱氮除硫菌的筛选鉴定及其生长特性研究

为促进反硝化脱硫工艺的工程化应用,从稳定运行的反硝化脱硫UASB中筛选出2株高活性自养反硝化菌H3和H7,并对其进行了鉴定和生长及反硝化特性的研究。结果表明,2株菌均为革兰氏阴性菌,16S r DNA序列分析表明,分别与Pseudomonas stutzeri CONC12和Pseudomonas stutzeri 19smn4相似性为99.6%和98.9%。结合生理生化特性和16S r DNA序列分析,确定两菌株都为Pseudomonas stutzeri。对两菌株的生长和反硝化特性研究表明,菌株H3和H7的生长最适起始p H为6.94和6.88,最适反硝化p H分别为6.77和6.56。H3和H7的最适生长温度为30.4℃和30.6℃,最适反硝化温度分别为31.4℃和31.2℃,两株菌种都为非耐盐菌株。     

酯酶产生菌的分离与酶学性质研究

从餐馆附近下水道收集到的土壤中分离获得6株酯酶产生菌,其中S2菌株活性最高,从其形态特征、生理生化试验以及16S rRNA序列分析等方面,初步鉴定S2菌株为假单胞属(Pseudomonas sp.)。对该菌株的部分酶学性质进行了研究,发现该菌株所产的酯酶最适反应温度为40℃,最适反应pH为8.0;且该酯酶在温度为60℃以下和pH 7.0～10.0具有良好的稳定性。     

一株阿维菌素降解菌AW1-18的筛选与分类鉴定

利用富集培养法从长期施用阿维菌素农药的菜豆土壤中分离出18株能利用阿维菌素为唯一碳源和氮源生长的细菌.采用紫外分光光度法对菌株的阿维菌素降解能力进行测定,结果有4株菌有较好的降解率,其中降解率最好的菌株命名为AW1-18.通过高效液相色谱法对AW1-18的降解能力进行测定,结果表明,当接种量为2.5％时,该菌株在含100 mg/L的阿维菌素无机盐基础培养液中,30℃,150 r/min,pH7的条件下培养6d后,对阿维菌素的降解率达75％以上.用16S rRNA通用引物,经PCR扩增、测序得到AW1-18的16S rRNA序列(GenBank登录号为JQ316540),在NCBI中通过BLAST后发现与不动杆菌属(Acinetobacter)的16S rRNA序列相似性达到99％,结合形态特征和生理生化特性分析结果,确定菌株AW1-18为Acinetobacter tandoii.     

一株丁草胺降解菌的分离鉴定及其降解特性的研究

利用富集培养技术从长期施用丁草胺的稻田土壤中分离得到能够降解丁草胺的细菌1株, 标记为LYC-1。经形态特征、生理生化特征和16S rRNA序列分析, 将该菌株鉴定为不动杆菌属(Acinetobacter sp.), 菌株LYC-1的最适生长温度为30°C, 最适pH值为7.5。当接种量为5%时, 该菌株在含100 mg/L 的丁草胺无机盐基础培养液中培养7 d后, 可使丁草胺降解达80%以上。     

Polyhydroxyalkanoate (PHA) production using waste vegetable oil by Pseudomonas sp. strain DR2

To produce polyhydroxyalkanoate (PHA) from inexpensive substrates by bacteria, vegetable-oil-degrading bacteria were isolated from a rice field using enrichment cultivation. The isolated Pseudomonas sp. strain DR2 showed clear orange or red spots of accumulated PHA granules when grown on phosphate and nitrogen limited medium containing vegetable oil as the sole carbon source and stained with Nile blue A. Up to 37.34% (w/w) of intracellular PHA was produced from corn oil, which consisted of three major 3-hydroxyalkanoates; octanoic (C8:0, 37.75% of the total 3-hydroxyalkanoate content of PHA), decanoic (C10:0, 36.74%), and dodecanoic (C12:0, 11.36%). Pseudomonas sp. strain DR2 accumulated up to 23.52% (w/w) of PHAMCL from waste vegetable oil. The proportion of 3- hydroxyalkanoate of the waste vegetable-oil-derived PHA [hexanoic (5.86%), octanoic (45.67%), decanoic (34.88%), tetradecanoic (8.35%), and hexadecanoic (5.24%)] showed a composition ratio different from that of the corn-oil-derived PHA. Strain DR2 used three major fatty acids in the same ratio, and linoleic acid was the major source of PHA production. Interestingly, the production of PHA in Pseudomonas sp. strain DR2 could not occur in either acetate- or butyrate-amended media. Pseudomonas sp. strain DR2 accumulated a greater amount of PHA than other well-studied strains (Chromobacterium violaceum and Ralstonia eutropha H16) when grown on vegetable oil. The data showed that Pseudomonas sp. strain DR2 was capable of producing PHA from waste vegetable oil.     

柴油降解菌的筛选及降解能力研究

从修造船业周围油污污染土样中分离纯化出9株以柴油为唯一碳源的高效降解菌,其中2~#菌为微球菌属,确定为优势菌株,其降解率高达到65.7%;分析了接种量、柴油浓度、pH、温度、转速对2~#微球菌降解柴油的影响.结果表明,该菌株最适宜生长条件接种量为1.0ml/L、柴油浓度为1.4g/L、pH7.0、温度为35℃、转速为160 r/min.     

原生质体紫外诱变选育可利用廉价碳源的高产油新菌株

【目的】研究并建立利用原生质体紫外诱变技术选育可利用廉价碳源发酵的高产油新菌株的方法。【方法】采用1.5%蜗牛酶和1.0%纤维素酶混合液水解去除细胞壁得到2A00015(近平滑假丝酵母,Candida parapsilosis)的原生质体,将其放于紫外灯下诱变及再生壁培养,筛选获得可利用廉价碳源发酵的高产油酵母,并采用气相色谱质谱联用法(GC-MS)测定其脂肪酸组成。【结果】突变效果最好的突变菌株2A00015/25用葡萄糖发酵培养7 d后,其生物量、油脂产率和产油量分别为17.77 g/L、58.12%和10.32 g/L,较原始菌株分别提高了12.45%、23.32%和38.68%;利用废糖蜜发酵培养,其生物量、油脂产率和产油量分别为18.54 g/L、49.44%和9.17 g/L,较原始菌株分别提高了9.09%、21.16%和32.18%。利用废糖蜜培养其产油效率虽低于利用葡萄糖培养,但从环境保护及原材料成本的角度考虑,用废糖蜜作为碳源发酵培养产生油脂更具优势。诱变菌株利用废糖蜜发酵后产生油脂经检测含有8种脂肪酸,其脂肪酸组成与植物油近似,其中不饱和脂肪酸含量占脂肪酸总量的82.4%。【结论】通过利用原生质体紫外诱变技术,成功选育出一株新的可利用廉价碳源的高产油海洋菌株,产油率达到49.4%,提高了21.2%。     

高产不饱和油脂海鞘真菌桔青霉Asc-2-4的诱变选育

诱变育种是获得高产菌株,实现微生物工业化生产油脂的重要措施。以前期获得的高产不饱和油脂菌株桔青霉（Penicillium citrinum）Asc-2-4为出发菌株,利用丙二酸建立快速筛选高产不饱和脂肪酸突变菌的方法,通过紫外线 氯化锂复合诱变得到1株高产油脂突变菌Asc-2-4-1,油脂含量比出发菌株提高了92.98%。经过初步的培养基无机盐优化,其油脂得率和不饱和脂肪酸产量达到了7.10 g/L和3.84 g/L,与Asc-2-4相比,分别提高了84.42%和77.78%。结果表明,通过复合诱变选育技术可选育出高产突变菌株,选育的Asc-2-4-1可望作为产油微生物被开发利用。     

Investigation of alkane biodegradation using the microtiter plate method and correlation between biofilm formation, biosurfactant production and crude oil biodegradation

Among 25 crude oil-degrading bacteria isolated from a marine environment, four strains, which grew well on crude oil, were selected for more study. All the four isolated had maximum growth on 2.5% of crude oil and strain BC (Pseudomonas) could remove crude oil by 83%. The drop collapse method and microtiter assay show that this strain produces more biosurfactant, and its biofilm formation is higher compared to other strains. Bacterial adhesions to crude oil for strains CS-2 (Pseudomonas), BC, PG-5 (Rhodococcus) and H (Bacillus) were 30%, 46%, 10% and 1%, respectively. Therefore, strain H with a low production of biosurfactant and biofilm formation had showed the least growth on these compounds. PCR analysis of these four strains showed that all isolates had alk-B genes from group (III) alkane hydroxylase. All isolate strains could utilize cyclohexan, octane, hexadecane, octadecan and diesel fuel oil; however, the microtiter plate assay showed that strain BC had more growth, respiration and biofilm formation on octadecan.     

Petroleum Degradation,Biosurfactant and Laccase Production by Fusarium neocosmosporiellum RH-10: A Microcosm Study

The aim of this study was to evaluate the potential of crude oil removal by fungal strains isolated from Arak refinery. The results showed that the RH10 strain is a potent strain as a surfactant producer and degrader of petrochemical hydrocarbons. The strain was identified as a Fusarium neocosmosporiellum and could degrade 58% of hydrocarbons in the minimal medium and reduce the surface tension from 45 to 26.5 mN m^?1. Moreover, residual crude oil analysis with Fourier transform infrared spectrophotometry showed that this strain was able to degrade 50% of aliphatic compounds. To investigate the mechanism of degradation, oxidase enzymes were assayed and it was found that F. neocosmosporiellum can produce 1.94 U L^?1 of laccase in 10 g L^?1 crude oil. Carbon, nitrogen, phosphorus and soil pattern optimization in a microcosm study showed that this strain removed 44% and 27% of the crude oil from contaminated soil in 1% and 5% crude oil concentrations, respectively. Under optimum condition, 9.66 g kg^?1 crude oil was removed by F. neocosmosporiellum when the initial oil concentration was 50 g kg^?1, at the end of 150 days microcosm experiment. The results demonstrated the promising potential of fungi strain for cleaning of contaminated soil.     

Screening and Evaluation of Biosurfactant-Producing Strains Isolated from Oilfield Wastewater

The six biosurfactant-producing strains, isolated from oilfield wastewater in Daqing oilfield, were screened. The production of biosurfactant was verified by measuring the diameter of the oil spreading, measuring the surface tension value and emulsifying capacity against xylene, n-pentane, kerosene and crude oil. The experimental result showed three strains (S2, S3, S6) had the better surface activity. Among the three strains, the best results were achieved when using S2 strain. The diameter of the oil spreading of the biosurfactant produced by S2 strain was 14 cm, its critical micelle concentration (CMC) was 21.8 mg/l and the interfacial tension between crude oil and biosurfactant solution produced by S2 strain reduced to 25.7 mN/m. The biosurfactant produced by S2 strain was capable of forming stable emulsions with various hydrocarbons, such as xylene, n-pentane, kerosene and crude oil. After S2 strain treatment, the reduction rate of oil viscosity was 51 % and oil freezing point reduced by 4 °C.     

卡门柏青霉-PG3脂肪酶的研究

从242株青霉属菌株中筛选出脂肪酶产生菌青霉-PG3。经鉴定,定名为卡门柏青霉(Penicillium camembertii Thom)。卡门柏青霉-PG3在由4％豆饼粉,0.5％糊精,0.75％橄榄油,0.5％K_2HPO_4,0.1％(NH_4)_2SO_4组成的液体培养基中,28℃,振荡培养96小时,发酵液脂肪酶活力(39℃,pH7.0)达60U/ml。PG3脂肪酶以橄榄油为底物,水解反应最适温度为48℃,最适pH为8.0。pH稳定范围6.0—11.0。Cu~(2+),Ca~(2+),Fe~(2+),Pb~(2+)等金属离子对酶活力有抑制作用。PG3脂肪酶对椰子油、菜籽油、亚麻油等油脂的水解率分别达到96％,94％和90％。     

以脂肪酶为指标筛选赤霉素高产菌株的研究

以藤仓赤霉菌978对978#菌丝机械断裂后,采用紫外(30W,20cm)诱变处理60s,以脂肪酶活性为初筛指标筛选得到4株产赤霉素效价比出发菌株高的突变株,其中菌株GL-2在添加3%豆油的发酵培养基中产赤霉素能力比菌株978#在全淀粉发酵培养基发酵产素能力提高了1.2倍.调整补油工艺后,菌株GL-2产素能力进一步提高到...     

The effect on some enzymes of rat tissue of diets low in fat content

1. Rats of two strains were kept on three different diets; one was a commercial diet of rat pellets, one contained about 80% of sucrose and 20% of casein and was supplemented with corn oil, and the third was a similar diet without the corn oil. 2. On the commercial diet, the specific activities of pyruvate kinase, glucose 6-phosphate dehydrogenase and fructose 1,6-diphosphatase in the livers of one strain of rats (strain A) were 1.5-3 times those in the other strain (strain B). When the diet high in sucrose and supplemented with corn oil was given, there were large increases in the specific activity of pyruvate kinase, glucose 6-phosphate dehydrogenase and fructose 1,6-diphosphatase in the livers of strain A rats. With strain B rats the increases were much smaller. Omission of corn oil from the diet caused a threefold increase in the specific activity of glucose 6-phosphate dehydrogenase in strain B rats, but had little effect on other enzymes. 3. The enzymes of the kidneys and hearts of strain A rats were also more active than those of strain B rats. In strain A rats, the specific activities of pyruvate kinase and fructose 1,6-diphosphatase in the kidney increased when the sucrose content of the diet was high, but in the kidneys of strain B rats there was little change. 4. In strain A rats, the specific activity of pyruvate kinase in the heart more than doubled with the high-sucrose-corn oil diet and increased threefold when corn oil was omitted. No changes were seen in strain B rats. 5. In strain A rats, omission of corn oil from the diet increased the ability of the kidneys to synthesize glucose from lactate. 6. In strain B rats, addition of corn oil to the diet resulted in a decrease in the liver in the specific activity of ATP citrate lyase and in the ability to incorporate acetate into lipid.