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1.
用PCR方法扩增了一株石油降解菌株G5的16S rRNA基因全序列,并对其进行了克隆和测序.对该序列在GenBank中的BLAST结果表明,所有与该序列高度同源的序列都是假单胞菌的16S rRNA基因.其中假单胞菌的代表菌株Pseudomonas aeruginosa,P.fluoroscens,P .putida,P.syringae的16S rRNA基因序列与G5的16S rRNA基因序列同源性分别为93.4%,98.4%,96.3%,97.5%.对G5和其他39株假单胞菌的16S rRNA基因序列进行聚类分析,获得的系统发育树与RDP(Ribosomal Database Project)报道的系统发育树基本一致,其中菌株G5与5株P.chlororaphis聚类在一起.图2参7  相似文献   

2.
为了解煤层中产甲烷菌的生理生化特性,结合厌氧培养箱和平板划线分离技术从山东兖煤菏泽能化公司赵楼煤矿距离地表936 m处坑道顶板取得40℃煤层水样品中微生物进行富集分离纯化.结果表明:在该条件下存在产甲烷微生物,并分离得到一株产甲烷菌株M-3,该菌株呈短杆状,菌体大小约(1.0-2.0)μm×0.5μm;革兰氏染色显阳性;在平板上生长出圆形黄色菌落,边缘光滑整齐;可以利用乙酸、甲酸、甲醇和H2+CO2(V/V=2:1)作为唯一碳源生长;最适生长温度为45℃;对酸碱具有良好的适应性,中性条件下甲烷产量最多;最适NaCl浓度为0.2-0.6 mol L-1.对菌株M-3的16S rRNA基因序列同源性分析表明该菌株与Methanobacterium bryantii同源性高达99%,G+C含量32.9%.本研究通过形态、生理生化特性以及16S rRNA分析,鉴定菌株M-3为M.brytantii.  相似文献   

3.
为促进嗜盐微生物及其所产蛋白酶的工业应用,采用脱脂奶粉培养基从我国天津近海盐田富集筛选得到产蛋白酶中度嗜盐菌株SY-7.通过对其形态特征、生理生化、16S rRNA基因序列及系统进化树进分析,确定菌株的分类.利用Plackett-Burman(PB)设计和响应面优化法(Response surface methodology)优化其发酵条件.结果表明:菌株SY-7为革兰氏阳性菌,最适生长温度为30℃、最适生长pH 7.5,最适NaCl为10%.16S rRNA基因序列分析显示,菌株SY-7与Thalassobacillus devorans亲缘性最近,16S rRNA基因序列相似性为98.88%.利用PB设计从众多影响因素中筛选出影响较大的3个因素:葡萄糖含量、接种量和装液量;再利用响应面法中的杂合设计进优化,通过拟合得到响应曲面函数,获得了最佳的产酶条件.在该实验条件下酶活从870 U/mL提高到1 390 U/mL,实际酶活力达到理论预测值的98.3%,优化效果较好.  相似文献   

4.
苯酚降解菌UW7的鉴定及对苯酚的降解作用   总被引:5,自引:0,他引:5  
从焦化厂污水处理曝气池泥样中分离出具有降解苯酚能力的UW7菌株,根据形态、生理生化性状初步鉴定为不动杆菌属(Acinetobacter).该菌16S rRNA基因序列(在GenBank中的登录号为GU083586)与多株鲁氏不动杆菌(A.lowffii)的相似性在99%以上.结合形态、生理生化特性,鉴定UW7菌株为鲁氏不动杆菌(A.lowffii UW7),该种细菌具有降解苯酚的特性尚未见报道.该菌株降解苯酚的最适温度为30℃,最适生长pH 7.0,对2.5 g/L浓度的苯酚能够有效降解,对3.5~4.0 g/L浓度的苯酚有较强的耐受能力,是处理高浓度苯酚废水的良好菌种资源.  相似文献   

5.
为开发极端环境工业用酶,从新疆盐碱土微生物中分离得到一株中度嗜盐高产淀粉酶活性菌株H3,其能耐受30%盐浓度和pH 11的极端环境.通过形态特征、生理生化实验及16S rRNA基因序列分析,确定H3属于Gracilibacillus属.该菌株能在盐浓度为0~30%的培养基上生长,最适生长盐浓度为5%~10%,最适pH值为8.5.在最适生长盐浓度、pH条件下,其淀粉酶活性可达到4 830个活力单位,可用于高盐高碱环境下淀粉的水解.  相似文献   

6.
萘降解菌NAP_A的分离、降解性能和分子系统学研究   总被引:2,自引:0,他引:2  
张春杨  彭振兴 《生态环境》2008,17(1):109-112
该文研究降解多环芳烃类环境污染物的微生物资源、降解活性和分子系统特征.利用萘-无机盐选择性培养基分离萘降解菌,用培养技术和气相色谱法检测菌株对底物的利用和降解情况,用分子克隆技术获得菌株的16S rRNA基因并测序,用DNAMAN软件对菌株的16S rRNA基因序列进行比对和系统发育分析.从淄博张店污水处理厂的活性污泥中分离到一株能降解萘的菌株NAP_A.此菌株在30 ~35 ℃和pH 7条件下较快的降解底物萘,其中30 ℃下,10 d内可以将初始质量浓度为320 mg·L-1的萘降解90%± 4.5%.对菌株NAP_A的16S rRNA基因进行了克隆和测序(EU142847),基于菌株NAP_A和相关菌株的16S rRNA基因进行系统发育分析,结果表明菌株NAP_A位于苍白杆菌属的分枝中,其中与假中间苍白杆菌种的同源性最高,可达99%,因此推断NAP_A菌是一株假中间苍白杆菌.此前并无苍白杆菌属成员降解多环芳烃的报道.这是首例苍白杆菌属成员降解多环芳烃的报道,对今后在环境污染防治中开发利用此类细菌具有指导意义.  相似文献   

7.
青海湖嗜盐菌的分离与优势菌株QHL5的特性   总被引:1,自引:0,他引:1  
采用高盐选择性培养基从20份青海湖水样中筛选分离获得35株青海湖嗜盐微生物.耐盐梯度实验分析表明:水体中以中度嗜盐菌为主,约占65.7%;弱嗜盐菌次之,约占25.7%;非嗜盐菌与耐盐菌相对较少,约占8.5%.其中优势菌QHL5的生长盐度耐受范围为0.04~3.0 mol/L,最适生长范围为0.3~0.9 mol/L,隶属于中度嗜盐菌.菌体形态和生长特性分析表明:QHL5菌落大小为4~6 mm,圆形或椭圆形,边缘整齐厚实,乳白色,中间微隆起,不透明,革兰氏染色阴性.显微形态呈杆状,最适生长pH为8.5~9.5,最适生长温度为35℃.生理生化、16S rRNA的全序列Blast和系统发育分析表明:QHL5菌株分类定位于γ变形菌纲海洋螺菌目盐单胞菌属樊氏盐单胞菌(Halomonas ventosae).通过80%乙醇法抽提胞内相溶物质,采用TLC法检测出该菌积聚四氢嘧啶.  相似文献   

8.
泸州古酿酒窖池中两株产甲烷杆菌比较研究   总被引:1,自引:0,他引:1  
采用厌氧操作技术,从泸州老窖古酿酒窖池窖泥中分离到两株产甲烷杆菌0372-D1和0072-D2.0372-D1菌体形态为长杆状,略弯,两端整齐,不运动,可由多个菌体形成长链;在固体培养基中难以长出菌落,只利用H2+CO2产生甲烷.0072-D2菌体形态为弯曲杆状,淡黄色圆形菌落,利用H2+CO2或甲酸盐作为唯一碳源生长.两株菌最适生长温度均为35℃、菌株0372-D1最适生长pH为6.5~7.0,生长pH范围5.0~8.0;菌株0072-D2最适生长pH则为7.5.在各自最适条件下培养,两株菌的最短增代时间分别为19 h和8 h.通过形态、生理生化特征和16S rDNA序列的同源性分析,表明菌株0372-D1为产甲烷杆菌属的一个新种.0072-D2则为甲酸甲烷杆菌(Methanobacterium formicicum)的新菌株,相似性为99%.  相似文献   

9.
采用改进的亨盖特(Hungate)厌氧技术,从西藏林芝厌氧消化系统中分离到一株产甲烷菌菌株LZ-6.该菌株为革兰氏阴性,不运动,球形,直径约0.3~0.6μm.该菌株利用H2/CO2,微利用甲酸生长,不利用乙酸、甲醇、甲醇/H2、三甲胺、甲胺,最适生长pH为6.8~7.2,最适生长温度25℃,最适Na+浓度0.2mol/L.菌株LZ-6的16SrRNA基因序列与小甲烷粒菌(Methanocorpusculum parvum)相似性为99%.生理、形态结构特征等生物学特性研究表明此株产甲烷古菌为兼性嗜冷产甲烷古菌.  相似文献   

10.
为揭示多环芳烃(Polycyclic aromatic hydrocarbons,PAHs)降解微生物资源的多样性和石油降解菌群的优势菌,研究了从南海沉积物中分离得到的一株PAHs降解菌D22F的降解特性及其在石油降解菌群D22-1中的生态位.对菌株D22F进行16S rRNA基因同源性分析及透射电镜观察以初步确定其种属,通过培养法、气相色谱质谱联用(GC-MS)测定其多环芳烃降解范围和降解率,通过简并引物PCR扩增其PAHs双加氧酶大亚基基因片段并进行系统发育分析,采用变性梯度凝胶电泳(DGGE)监测石油降解菌群中的优势菌.结果表明,与菌株D22F的16S rRNA基因相似度最高的模式株为产卟啉杆菌属Porphyrobacter tepidarius DSM 10594T(AF465839;98.55%).该菌株能降解萘、甲基萘、苊、硫芴、菲、蒽等;对初始浓度为0.2 g/L菲10 d后的降解率可达90%以上.从其基因组DNA中克隆到的PAHs起始双加氧酶大亚基基因phnAc与Novosphingobium aromaticivorans DSM 12444中的质粒pNL1(CP000676)上的bphA1f基因相似度最高,达到99.41%.DGGE谱图显示,菌株D22F是石油降解菌群中的3种优势菌之一,在传代菌群中可稳定存在.Porphyrobacter sp.D22F为产卟啉杆菌属(Porphyrobacter)中首株以低分子量PAHs为唯一碳源和能源的菌株,是石油降解菌群的优势菌.  相似文献   

11.
The total lipid and wax ester content as well as the fatty acid and alcohol composition of all copepodid stages and adults of Calanus finmarchicus s.l. were investigated at different locations in the North Sea in 1983 and 1984. Total lipid and the wax ester proportion increased exponentially until Copepodid V. The females were sometimes lower in lipids than the Stage V. The wax ester proportion reached about 90% of total lipids in males and Copepodid V and up to 40% in Copepodid I. The major fatty acids were 16:0, 20:5, and 22:6 and the major fatty alcohols were 16:0, 20:1 and 22:1. At one station the 18:4 acid became one of the dominant acids, because of a Phaeocystis sp. bloom, indicating that the fatty acids of the diet are incorporated mostly unchanged into the lipids of the copepods. The other main fatty acids 20:1 and 22:1 are probably synthesized de novo, serving as precursors for the principal alcohols 20:1 and 22:1. Their levels decreased in the younger stages due to increases in 16:0 alcohol. The fatty alcohol-forming enzyme seems to be specific for saturated and monounsaturated acids, which may be synthesized de novo or derived from diet.  相似文献   

12.
Lipid compositions of the dominant Antarctic copepods Calanoides acutus, Rhincalanus gigas and Calanus propinquus from the Weddell Sea have been investigated in great detail. Copepods were collected during summer in 1985 and late spring/early winter in 1986. The analyses revealed specific adaptations in the lipid biochemistry of these species which result in very different lipid components. The various copepodite stages of C. acutus synthesize wax esters with long-chain monounsaturated moieties and especially the alcohols consisted mainly of 20:1(n-9) and 22:1(n-11). R. gigas also generates wax esters, but with moieties of shorter chain length. The fatty alcohols consisted mainly of 14:0 and 16:0 components, while the major fatty acids were 20:5, 18:4 and 22:6, of which 18:4 probably originated from dietary input. In contrast, C. propinquus accumulates triacylglycerols, a very unusual depot lipid in polar calanoid copepods. Major fatty acids in C. propinquus were the long-chain monounsaturates 22:1(n-9) and 22:1(n-11), which may comprise up to 50% of total fatty acids. In C. acutus and C. propinquus there was a clear increase of long-chain fatty acids with increasing developmental stage. In contrast, the fatty acid and alcohol composition of the R. gigas copepodite stages were characterized by the dominance of the polyunsaturated fatty acids as well as high amounts of the monounsaturates 18:1(n-9) and 16:1(n-7). There was a considerable decrease of the dietary fatty acid 18:4(n-3) towards the older stages during summer; in late winter/early spring 18:4 was only detected in very low amounts. This tendency was also found in the other two species, but was less pronounced. In all three species dry weight and lipid content increased exponentially from younger to older stages. The highest portion of wax esters, or of triacylglycerols in C. propinquus, was found in the adults. Dry weight and lipid content were generally higher during summer. In late winter/early spring the variability was more pronounced and lipid-rich specimens showed a selective retention of long-chain monounsaturated fatty acids, whereas in lipid-poor specimens these fatty acids were very much depleted.  相似文献   

13.
The variation in the concentration and fatty acid composition of lipid classes during the molting cycle of the prawn Penaeus japonicus was investigated. The lipid concentration of the whole body reached a maximum at mid-premolt (Stage D2) and then decreased to low level at late premolt (Stage D3–4). The accumulation of lipids during the premolt period seemed to be attributable to the increase of both polar and neutral lipids. The increase of neutral lipids at Stage D2 was derived from not only triglycerides but also free sterols and free fatty acids. Regarding the fatty acid composition of every lipid class, a marked variation occurred mainly at the intermolt (Stage C). In this stage, the polar lipids were rich in monoenoic acids such as 18:1 and poor in polyenoic acids such as 20:53 and 22:63. The triglycerides were rich in polyenoic acids at Stage C, but poor in monoenoic acids such as 16:1 and 18:1. The steryl esters contained large amounts of saturated acids such as 16:0 and 18:0 throughout the molting cycle, however the level of polyenoic acids increased at Stage C.  相似文献   

14.
The fatty acids of two composts of active sludge with palm tree waste were investigated by thermochemolysis coupled to gas chromatography-mass spectrometry. This method (tetramethylammonium hydroxide-pyrolysis-gas chromatography-mass spectrometry) allowed the direct determination of total fatty acids (analysed as fatty acid methyl esters: FAMEs) present in the organic matter of the samples without any separation procedures. Mixture A was 1/3 sludge+2/3 palm waste and mixture B was 1/2 sludge+1/2 palm waste. The level of FAMEs rose by 8.4–33.3% and 10.8–13.4% in mixtures A and B, respectively, after 6 months of co-composting. Branched FAMEs of bacterial origin (iC15:0) rose during the thermophilic phase, in mixture A the aC17:0/aC15:0 ratio increased during the co-composting process, also in mixture B the aC16:0/C16:0 ratio rose but only during the thermophilic phase. All the FAMEs identified showed a drop at the end of co-composting except for C18:0 and C16:0. The stabilisation phase was characterised by a significant rise in the length of the aliphatic chains; the carbon preference index thus increased at the end of the composting process, indicating that the final product was proportionally richer in fatty acids of plant origin.  相似文献   

15.
Surface sediments from the Svartnes basin (195 m deep) in Balsfjorden, northern Norway (ca. 70°N), were partially characterized to assess the nature and origin of the organic material present and its potential nutritive value for sediment-ingesting animals. Seasonal analyses were carried out on material collected between May 1979 and August 1980 for total organic matter, organic C and N, acid-extractable amino acids and lipids extractable with chloroform:methanol. Little or no seasonal variation was seen in any of the parameters analysed. The mean apparent organic content was 9.3% of sediment dry weight, the organic C content was 2.38%, the organic N content was 0.26% and the C:N ratio was 9.1:1. Acid hydrolysis of sediment yielded 575 mg of amino acids and 41 mg of NH3 per 100 g sediment dry weight, the composition of the amino acids being similar to that of nutritionally highquality animal or microbial protein. Glucoseamine was not detected in acid hydrolysates of sediment, consistent with the absence of chitin. Chloroform:methanol extraction yielded 133 mg of material per 100 g sediment dry weight, 62% of which was accounted for by saponifiable lipids (fatty acids) and non-saponifiable lipids present in approximately equal amounts. Fatty alcohols accounted for 30% of the non-saponifiable lipids and phytol accounted for 40% of the fatty alcohols. Small amounts of very long-chain fatty alcohols characteristic of terrestrial plants were present, but long-chain monounsaturated fatty alcohols characteristic of marine zooplankton were essentially absent. Very small quantities of long-chain ketones characteristic of marine coccolithophores were detected. The major fatty acids present in sediments were 16:0, 14:0, 16:1 (n-7) and 18:1 (n-9), and 11% of the total fatty acids were comprised of a mixture of odd-numbered straight-chain and branched-chain moieties characteristic of micro-organisms. The data point to small amounts of material characteristic of marine and terrestrial photosynthetic organisms being present in sediments at any given time, whereas material characteristic of marine zooplankton is not present. The presence of material characteristic of micro-organisms is consistent with conversion of sedimenting material into a pool of sediment micro-organisms. Although the biological availability to sediment-ingesting organisms of the esterified fatty acids and the acid-extractable amino acids in sediments remains to be assessed, the maximum amounts available are equivalent to approximately 60 mg of polar lipid and 575 mg of protein per 100 g sediment dry weight. These amounts are small in relation to the production rates in the photic zone of the fjord, implying that the bulk of the energy flow in Balsfjorden occurs in its relatively short pelagic food chains.  相似文献   

16.
二十二碳六烯酸(Docosahexaenoic acid,DHA C22:6n-3)是具有各种重要生理功能的高度不饱和脂肪酸.分别以质粒pYTFD5和pYFAD4为模板,扩增获得860 bp的△5-延长酶基因(elo5)和1 600 bp的△4-脱饱和酶基因(fad4).利用重叠延伸PCR构建elo5-fad4融合基因,Hind Ⅲ/Sph Ⅰ双酶切后连接到经同样处理过的pYES2.0载体,构建重组表达质粒pYEL05-FAD4.转化酿酒酵母尿嘧啶缺陷型菌株INVScl,通过缺少尿嘧啶的选择性培养基筛选阳性克隆子.添加外源脂肪酸C20:5底物.半乳糖诱导表达.气相色谱-质谱分析表明,重组酵母总脂肪酸中出现了DHA(二十二碳六烯酸,C22:6n-63)新产物,融合基因elo5-fad4在酿酒酵母中得到了表达.图7表1参18  相似文献   

17.
Mid-water, oceanic crustaceans were either fed food labelled with (14C) palmitic acid under controlled conditions, or injected directly with (14C) palmitic acid. Lipid classes and their constituent fatty acids and fatty alcohols were subsequently separated and assayed for radioactivity. Significant levels of radioactivity were present in the 16:01 alcohols and 18:1 acids of wax esters, and in the 16:0/16:1, 18:0/18:1, 20:5 and 22:6 acids of both triglycerides and phospholipids. It was concluded that these crustaceans were capable of biosynthesis of wax esters and higher polyunsaturated fatty acids.  相似文献   

18.
Carbon assimilation and lipid production were studied in phytoplankton in Balsfjorden and Ullsfjorden, northern Norway, during the exponential growth phase of the spring bloom in 1983 (6–7 April). In Balsfjorden, phytoplankton biomass was constant with depth and equivalent to 1.5 g chlorophyll a 1-1. Phytoplankton biomass in Ullsfjorden varied with depth, with a maximum of ca. 7 g chlorophyll a 1-1 occurring at 5 to 10 m. Particulate carbon-14 assimilation was about 18 mg C per m-2 h-1 in Balsfjorden and about 39 mg C per m-2 h-1 in Ullsfjorden over the depth range 4 to 8 m. In Balsfjorden, the percentage of total fixed carbon recovered as total lipid was 14.7 and 20.4% at 4 and 8 m depth, respectively. In Ullsfjorden, the corresponding values were 8.8 and 28.1% at 4 and 8 m, respectively. The percentages of total fixed carbon present as fatty acids were 1.1 and 1.6% at 4 and 8 m, respectively, in Balsfjorden, and 0.8 and 6.4% at 4 and 8 m in Ullsfjorden. The majority of the radioactivity in lipid at both locations and at both depths was present as polar lipid, with small percentages present in triacylglycerols and very small percentages present in free fatty acids. On average, about 18% of the total carbon-14 incorporated into phytoplankton over a 6 to 7 h mid-day period was recovered as total lipid and its percentage tended to increase with depth. The relatively low percentage of incorporated carbon-14 present as fatty acids in total lipid implies that most of the radioactivity is present in glyceryl and/or glucosyl moieties and that measurement of total radioactivity in total lipid does not necessarily give an accurate estimation of lipogenesis in phytoplankton. Fatty acid analyses of total phytoplankton in Balsfjorden and Ullsfjorden in 1983 and of a surface slick at the end of a bloom of Phaeocystis pouchetii in Balsfjorden in May 1980 showed an abundance (more than 40% of the total) of (n-3) polyunsaturates in all cases. C-18 polyunsaturates, especially 18:4 and 18:5, were very abundant (about 30% of the total) in the P. pouchetii surface slick in Balsfjorden in 1980. Both P. pouchetii biomass and C-18 polyunsaturates were more abundant in Ullsfjorden than in Balsfjorden (1983). Lipids from the P. pouchetii surface slick were deficient in C-16 polyunsaturates and relatively deficient in C-20 polyunsaturates, but both these classes of fatty acids were abundant in Balsfjorden and Ullsfjorden in 1983. The phytoplankton in both locations in 1983 was dominated by P. pouchetii and diatoms; Chaetoceros socialis was especially abundant in Balsfjorden. The results are discussed in terms of the fatty acids present in herbivorous zooplankton in northern Norwegian fjords.  相似文献   

19.
Anthopleura elegantissima containing zooxanthellae, as well as isolated zooxanthellae, incubated with acetate-1-14C under both light and dark conditions readily incorporate radioactivity into their total lipid pools. In both cases, the specific activity was greatly increased in the light. Dark-incubated anemones and isolated zooxanthellae incorporate activity predominantly into polar lipid; the remainder being present principally in the triglyceride moiety. Light-incubated organisms, however, show a dramatic redistribution of isotope towards greatly increased triglyceride and was ester incorporation, with a concomitant drop in polar lipid. onder light conditions, 70 to 75% of the radioactivity found in the fatty acids of the total zooxanthellae lipid was present in hexadecanoic (16:0) and octadecenoic (18:1) fatty acids. These are also the two major fatty acids by mass. Octadecanoic acid (18:0) is less than 5% by mass. Isotope incorporation patterns suggest that octadecenoic acids arise by elongation of hexadecenoic acids and that this conversion is blocked in the dark. Isotope incorporation patterns for anemones suggest that fatty acids, primarily in the form of saturated or monoenoic fatty acids, are translocated from algal to animal cells. No activity was found in either octadecadienoic (18:2) or octadecatrienoic (18:3) acids. The significance of these data is discussed.  相似文献   

20.
This study investigated incorporation and metabolism of saturated [(1-14C) 16:0] and unsaturated [(1-14C) 18:26 and (1-14C) 18:33] fatty acids in adult eastern oysters,Crassostrea virginica Gmelin (spawned from parents obtained in 1986 from Mobjack Bay, Virginia, USA), and the influence of temperature on these processes. InC. virginica, incorporation of injected palmitic (16:0) and linolenic (18:33) acids was increased when oysters which had been grown in warm water (22 to 23°C) were transfered to cold water (5 to 7°C) for 8 to 18 d. Incorporation of linoleic acid (18:26) was unchanged under these conditions. The changes in concentration may have been linked to depression of metabolism in these oysters, in particular that of 16:0, which was reduced by 90%. Oxidation of incorporated fatty acids was much higher in warm than in cold water. Cold-temperature conditioning ofC. virginica altered the distribution of fatty acids among the neutral and polar lipid fractions. Long-term exposure to cold water increased the proportion of fatty acids in the polar fraction, which may be related to maintenance of membrane fluidity. Short-term exposure to cold water had the opposite effect, which may be due to increased energy requirements as the oyster adapts to new conditions. Reutilization of14C-acyl groups demonstrated de novo synthesis of 16:0 and 18:0 fatty acids. Only limited elongation and no desaturation of the administered fatty acids was observed.  相似文献   

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