龙须藤叶粗提物的分离纯化及生物活性初探

为进一步研究和开发新植物源农药,拓宽龙须藤(Bauhinia championii)的生物活性研究,探索其不同组分潜在的杀菌和除草活性,该研究通过常温冷浸提取法提取、真空浓缩得到甲醇提取物。结果表明:用硅胶柱层析分离纯化,经TLC检识和碘缸显色后整合得到9个组分。反复重结晶7号和8号组分中析出的物质,经TLC检识和测熔点,得到1个纯化合物,编为33号,经波谱数据分析与molbase库对照,鉴定该化合物为(1R,2S,3S,4S,5S,6S)-6-甲氧基-1,2,3,4,5-环己烷五醇,是一种重要的工业原料。杀菌和除草活性试验结果显示,粗提物在1 000μg·m L~(-1)时对水稻稻瘟病菌的抑制率为(40.84±1.00)%,对稗草根的抑制率为(49.18±2.33)%;各组分在500μg·m L~(-1)时,4号组分对水稻稻瘟病菌的抑制率达到(44.19±0.76)%,2号和3号组分对稗草根的抑制率分别为(88.92±1.31)%和(90.99±1.45)%,3号和6号组分对马齿苋根的抑制率分别为(72.06±1.31)%和(89.92±1.73)%。这表明龙须藤叶提取物对水稻稻瘟病菌、稗草根和马齿苋根有良好的抑制效果,可进一步分离2号、3号、4号、6号组分,以获得高活性的单体化合物。     

一组降解纤维素细菌的分离筛选及产酶特性研究

用稀释法从腐殖泥中分离菌株,根据它们在羧甲基纤维素钠(CMC-Na)-刚果红培养基上的透明圈直径及对滤纸的崩解能力,获得7株纤维素分解细菌(编号:B-37、B-35、B-31、B-25、B-17、Z-a和Z-b),其中菌株Z-b的纤维素崩解能力最强,分子鉴定结果表明它与Stenotrophomonas maltophilia的16S rDNA序列有99.8%的同源性,初步确定为嗜麦芽窄食单胞菌。适合7个菌株生长的C源为马铃薯浸出液,无机盐组分为:CaCl_2 0.20、MgSO_4 1.25、NaCl 5.00、(NH_4)_2SO_4 1.30、KH_2PO_4 1.35、FeSO_4·7H_2O 0.015、Na-EDTA 0.02g/L。在滤纸为唯一碳源的培养基中,菌株Z-b的滤纸酶(FPase)和CMC酶(CMCase)活性最大,为0.099 U/mL和0.075 U/mL,而在固体PSA上,菌株B-31和B-37的FPase和CMCase活性最高,为0.131 U/mL和0.175 U/mL。7个细菌单独发酵,Z-b对滤纸的崩解能力最强,滤纸块完全崩解成粉未状;与真菌34混合发酵,菌株组合Z-a+34、Z-b+34、B-31+34、B-25+34将滤纸完全水解为水溶性物质。可见,各菌株的纤维素酶活与培养条件密切相关,某些真菌、细菌间存在协同作用,它们混合发酵可大大提高纤维素的水解效率。     

南方红豆杉内生真菌的分离鉴定及其细胞毒活性研究

采用平板培养法对采自广东省的南方红豆杉进行内生真菌的分离,并通过形态学和分子生物学方法进行分类鉴定;共分离鉴定内生真菌22株,主要为刺盘孢属(Colletotrichum)、球座菌属(Guignardia)、座腔菌属(Botryosphaeria)、节菱孢属(Arthrinium)、炭角菌属(Xylaria)、毛壳菌属(Chaetomium)、交链孢属(Alternaria)、拟茎点霉属(Phomopsis)、长蠕孢属(Helminthosporium)等属。还采用MTT法测定了这些内生真菌发酵粗提物的细胞毒活性,活性研究发现有6个菌株的粗提物在作用浓度为100μg/mL时,对至少1种受试肿瘤细胞株的抑制率在50%以上,其中菌株A223和A240对人乳腺癌细胞MCF-7的抑制率在90%以上。表明南方红豆杉内生真菌多样性丰富,有些菌株具有良好的细胞毒活性,值得进一步深入研究。     

黄蜻幼虫肠道分离菌QTYC38活性代谢产物的分离和鉴定

【目的】黄蜻幼虫肠道分离菌QTYC38菌株的鉴定及抗菌除草活性代谢物的研究。【方法】通过形态学观察及分子生物学ITS序列分析,对菌株QTYC38进行鉴定。利用生长速率法和琼脂扩散法测定菌株粗提物及其单体化合物的抗菌活性,结合培养皿生物分析法测定菌株粗提物及其单体化合物的除草活性。同时运用多种色谱方法分离发酵产物中的活性成分,并根据质谱和核磁共振谱数据确定其结构。【结果】菌株QTYC38被鉴定为浅黄新萨托菌Neosartorya aureola,在供试浓度为100μg/mL时,其粗提物对稗草和反枝苋的生长抑制活性较好,抑制率均大于65%;当供试浓度为30μg/滤纸片时,其对金黄色葡萄球菌(Staphyloccocus aureus)具有较好的抑菌活性,抑菌圈直径为22.7 mm,与阳性对照药(硫酸庆大霉素23.2 mm)活性相当。从该菌固体发酵产物中分离纯化得到4个单体化合物:helvolic acid(1),aromatic lactones(2),questin(3)和erogosterol(4)。其中,化合物1对枯草芽孢杆菌(Bacillus subtilis)和金黄色葡萄球菌(S.aureus)均具有较好的生长抑制活性,最低抑菌浓度分别为3.1和1.5μg/mL;当供试浓度为100μg/mL时,化合物3和化合物4分别对杨树溃疡病菌(Dothiorella gregaria)和小麦赤霉病菌(Fusarium graminearum)具有较好的抑制效果,抑制率分别为52.4%和72.3%。【结论】菌株QTYC38具有开发为微生物源除草剂和新型杀菌剂的潜能。     

叶柄粘球菌STXZ77的分离鉴定及抗肿瘤活性

目的：从土样中分离纯化粘细菌,对其进行鉴定与归类,以丰富粘细菌菌种资源,并对其进行抗肿瘤活性初步研究,为抗肿瘤药物开发奠定基础。方法：采用灭活大肠杆菌诱导法,从土样中分离纯化粘细菌,结合形态观察、生理生化特征及16S rRNA基因序列同源性分析进行菌株鉴定;向发酵液上清中加入XAD-16大孔吸附树脂提取发酵产物粗提物,CCK-8法进行体外抗肿瘤活性研究;RP-HPLC分离抗肿瘤活性组分,LC-MS/MS分析其分子质量。结果：分离并鉴定了STXZ77菌株,命名为Myxococcus stipitatus STXZ77。该菌株的发酵产物XAD-16树脂粗提物对小鼠黑色素瘤细胞B16、小鼠乳腺癌细胞4T1、人肝癌细胞SMMC-7721、人宫颈癌细胞HeLa、人结肠癌细胞SW480等多种肿瘤细胞具有较好的细胞毒性,作用24h的IC₅₀值分别为5.34μg/ml、13.50μg/ml、11.93μg/ml、28.70μg/ml、48.09μg/ml,而对正常细胞人脐静脉血管内皮细胞HUVEC的毒性较小,IC₅₀值为17.09μg/ml,小于B16、4T1及SMMC-7721的细胞毒性。RP-HPLC分离得到抗肿瘤活性组分AP-C,质谱分析其分子质量为422.99m/z。结论：从土样中分离得到粘细菌Myxococcus stipitatus STXZ77,从该菌中分离得到抗肿瘤活性组分AP-C,具有开发成抗肿瘤药物的潜在价值。     

增强UV-B辐射和接种稻瘟病菌对2个水稻品种幼苗叶片3个生理指标的影响

通过盆栽实验研究了单一增强UV-B辐射(2.5、5.0和7.5 kJ·m-2·d-1)以及增强UV-B辐射和稻瘟病菌(Magnaporthe grisea,菌株Y98-16T和Y99-63C)侵染复合胁迫对水稻(Oryza sativa L. )品种'黄壳糯'('Huangkenuo')和'合系41'('Hexi-41')幼苗叶片相对电导率、MDA含量和SOD活性3个生理指标的影响.结果表明:在低剂量(2.5和5.0 kJ·m-2·d-1)单一UV-B辐射条件下,2个水稻品种幼苗叶片的3个生理指标均较对照(自然光)有所降低;而在高剂量(7.5 kJ·m-2·d-1)单一UV-B辐射条件下,2个水稻品种幼苗叶片的3个生理指标均较对照有所增加.与对照(仅接种稻瘟病菌)相比,经低剂量(2.5和5.0 kJ·m-2·d-1) UV-B辐射后再接种稻瘟病菌,'黄壳糯'幼苗叶片的3个生理指标总体上均降低;而经高剂量(7.5 kJ·m-2·d-1)UV-B辐射后再接种稻瘟病菌,'黄壳糯'幼苗叶片的3个生理指标均有所增加.经不同剂量UV-B辐射后再接种稻瘟病菌,'合系41'幼苗叶片的3个生理指标总体上均有所增加,且2个水稻品种幼苗叶片的相对电导率与SOD活性均有显著的正相关性(P<0.05).与对照(仅接种稻瘟病菌)相比,接种稻瘟病菌后再经不同剂量UV-B辐射处理,'黄壳糯'幼苗叶片的3个生理指标均有所增加;而 '合系41'幼苗叶片的3个生理指标变化各异.接种稻瘟病菌菌株Y98-16T后再用UV-B辐射处理,2个水稻品种幼苗叶片的3个生理指标间均有显著或极显著的正相关性(P<0.05或P<0.01);而接种稻瘟病菌菌株Y99-63C后再用UV-B辐射处理,2个水稻品种幼苗叶片的3个生理指标间无明显的相关性.研究结果显示,在增强UV-B辐射和稻瘟病菌侵染复合胁迫条件下,因2个胁迫因子的作用顺序不同、稻瘟病菌菌株不同、UV-B辐射量不同以及水稻品种的差异,水稻幼苗叶片的细胞膜透性、MDA含量和SOD活性呈现出不同的变化趋势.     

云南稻瘟病菌系谱与致病型的关系

为探究稻瘟病菌无性世代DNA水平的变异,寻找云南稻瘟病菌谱系(genetic lineage,G)和致病型之间的对应关系,根据稻瘟病菌散布的重复序列Pot2(Pyricularia oryzac transposon),对云南水稻主产区稻瘟病菌菌株DNA进行了rep-PCR(repetitive polymerase chain reaction)扩增,获得rep-PCR指纹。聚类分析将134个稻瘟病菌代表菌株划分为G1～G8等8个谱系,揭示云南水稻主产区稻瘟病菌无性系丰富的遗传多样性。进一步接种分析了8个谱系的29个稻瘟病菌菌株对33个云南主产区水稻品种的亲和性,依其毒性谱,采用STATISTICAL5．0软件的UP-GMA程序进行聚类分析,将其划分为P1～P6等6个致病型群(pathotype group)。结果表明同一谱系的稻瘟病菌菌株多数对应2～3个致病型群,少数1个或4个致病型群;但G1～G8等8个谱系中的部分菌株都可对应致病型群P2。因此,云南水稻主产区稻瘟病菌谱系和致病型群之间属于复杂关系类型。此外,33个水稻品种中的合系16和京国92抗全部29个稻瘟病菌株,云粳20和合系30对全部供试菌株表现感病,这对云南水稻主产区品种布局提供了稻瘟病抗性方面的依据。因此,从育种应用和生产实际需要出发,水稻品种抗瘟谱测定仍然必要。     

番茄内生菌St24的鉴定及其对灰霉病的生防作用

从番茄植株根茎部分离到1株有抑菌活性的植物内生放线菌菌株St24,对其分类地位以及对灰霉病菌的防治效果进行研究.结果表明: 菌株St24为酒红链霉菌.St24发酵液经石油醚萃取得到的粗提物对多种病原菌有抑制作用,其中对灰霉病菌的抑制作用最强,抑制菌丝生长的EC₅₀为11.78 mg·L^-1.粗提物处理灰霉菌后,菌丝量减少,菌丝体皱缩、断裂、原生质外渗,菌体细胞表面有许多瘤状畸形.处理后的灰霉病菌培养液在260 nm处比对照多出现一吸收峰,说明粗提物对病原菌的细胞膜透性有影响.经盆栽试验, St24发酵液对番茄灰霉病有保护和治疗的作用,100 mg·L^-1粗提物叶面喷雾的保护作用效果最好,24 h后防效达到94.3%,120 h后为85.4%.     

内生黑曲霉JS-1对稻瘟病的拮抗作用研究

利用平板对峙法和牛津杯法,从疏花水柏枝、金银花、秋华柳的内生菌中,筛选出1株对稻瘟病菌具有很强抑制作用的菌株JS-1。经生理生化实验和18S rDNAITS序列分析,确定该菌株为黑曲霉(Aspergillus niger)。实验结果表明,JS-1发酵液作用稻瘟病菌后,稻瘟病菌的菌丝变细,分支减少,菌丝基质颜色变浅,作用72 h后干重显著降低。进一步实验表明,该菌产生的活性物质位于其发酵液的乙酸乙酯酯相部位,对稻瘟病病菌抑制率高达96.1%。大田实验数据（天然接种圃）显示,添加该物质后,丰两优4号（中感）和广陆矮4号（易感）叶瘟病情指数分别只有16.25%和32.48%,对稻瘟病的防治取得了很好的效果,说明该菌株具有开发成高效生物农药的巨大潜能。     

桦褐孔菌活性物质的提取工艺及体外抗糖尿病活性

对桦褐孔菌活性物质的提取工艺及其体外抗糖尿病活性进行研究。桦褐孔菌子实体用乙醇浸提后,乙醇粗提物用不同有机溶剂萃取,醇提残渣再以热水浸提,用标准曲线法测定活性组分中活性成分的含量,并检测活性物质对羟基自由基(·OH)、超氧阴离子自由基(O2-·)的清除效果以及对关键糖代谢酶α-淀粉酶、α-葡萄糖苷酶的抑制作用。结果表明:4种活性组分(乙酸乙酯相、正丁醇相、水相和粗多糖)对羟基自由基(·OH)和超氧阴离子自由基(O2-·)都有较强的清除作用,其中乙酸乙酯组分的活性最高,乙醇粗提物萃取组分对α-淀粉酶有抑制活性,而粗多糖对α-葡萄糖苷酶有抑制作用;桦褐孔菌具有抗氧化和抗糖尿病活性,其活性与活性物质种类及其含量具有相关性。     

海洋芽孢杆菌(Bacillus marinus)B-9987菌株抑制病原真菌机理

[目的]探讨海洋芽孢杆菌(Bacillus marinus)B-9987菌株的代谢产物BMME-1,对植物病原真菌茄链格孢菌的抑菌作用机理.[方法]分别使用分光光法、气相色谱-质谱GC-MS联用技术、红外光谱法等,检测了BMME-1处理病原真菌后,菌体渗透性、细胞壁及细胞膜成份的变化.[结果]BMME-1对茄链格孢菌的抑菌中浓度(MIC_(50))为6.2 mg/L,最小杀菌浓度(MFC)为50 mg/L,在MIC_(50)浓度或高于此浓度处理靶标菌,将导致菌体蛋白质、核酸等大分子物质的外流;处理菌株葡聚糖结构β-型糖苷键、碳-氧键(C-O)、碳-氢键(C-H)等基团的特征吸收强度降低,-OH、C=O的伸缩振动吸收强度升高;菌体细胞壁几丁质结构中酰胺I键吸收强度发生变化;与对照菌株的麦角甾醇含量(62.52±3.31%)相比,处理菌株麦角甾醇减少为(56.36±2.52)%,同时出现麦角固醇合成中间产物粪甾醇.[结论]BMME-1对病原真菌的抑制表现为:干扰细胞膜麦角甾醇的合成从而改变了细胞的通透性;对细胞壁葡聚糖结构的影响较大而几丁质次之.     

Mycosporine-like amino acid (MAAs) production by Heterocapasa sp. (Dinophyceae) in indoor cultures

The possibility of using mycosporine-like amino acids (MAAs), with an apparent sunscreen function in nature, as ultraviolet radiation (UVR) blockers to prevent skin injury has been raised by diverse authors. Production of MAAs by the dinoflagellate Heterocapsa sp. (Dinophyceae) is shown here. Three major peaks with absorption maxima at 330.8, 332.0 and 333.2 nm were detected by high performance liquid chromatography (HPLC) analysis of methanolic extracts in all tested conditions. Analysis of crude extract by mass spectroscopy with electrospray ionization (MS-EI) showed a set of molecular ions ([M+H](+)) with main peaks being at m/z 242.4, 288.4, 303.3 and 333.3 u.m.a. According to these data, along with retention times, the MAA profile of Heterocapsa sp. is assumed to be composed of shinorine (lambda(max)=334 nm), mycosporine-2-glycine (lambda(max)=331 nm) and palythinol (lambda(max)=332 nm). A constitutive MAA content of about 4 microg (10(6) cells)(-1) was measured under exposure to PAR only. A maximal accumulation of MAA per culture volume of 1.1 mg l(-1) was obtained after 72 h of exposure to PAR+UVA, while the highest production rate (0.025 mg l(-1) h(-1)) was computed after 24 h of exposure to PAR+UVA+UVB.     

Genetically engineered biosynthesis of macrolide derivatives including 4-amino-4,6-dideoxy-L-glucose from Streptomyces venezuelae YJ003-OTBP3

Two sugar biosynthetic cassette plasmids were used to direct the biosynthesis of a deoxyaminosugar. The pOTBP1 plasmid containing TDP-glucose synthase (desIII), TDP-glucose-4,6-dehydratase (desIV), and glycosyltransferase (desVII/desVIII) was constructed and transformed into S. venezuelae YJ003, a strain in which the entire gene cluster of desosamine biosynthesis is deleted. The expression plasmid pOTBP3 containing 4-aminotransferase (gerB) and 3,5-epimerase (orf9) was transformed again into S. venezuelae YJ003- OTBP1 to obtain S. venezuelae YJ003-OTBP3 for the production of 4-amino-4,6-dideoxy-L-glucose derivatives. The crude extracts obtained from S. venezuelae ATCC 15439, S. venezuelae YJ003, and S. venezuelae YJ003-OTBP3 were further analyzed by TLC, bioassay, HPLC, ESI/MS, LC/MS, and MS/MS. The results of our study clearly shows that S. venezuelae YJ003-OTBP3 constructs other new hybrid macrolide derivatives including 4-amino-4,6-dideoxy-L-glycosylated YC-17 (3, [M+ Na+] m/z=464.5), methymycin (4, m/z=480.5), novamethymycin (6, m/z=496.5), and pikromycin (5, m/z=536.5) from a 12- membered ring aglycon (10-deoxymethynolide, 1) and 14-membered ring aglycon (narbonolide, 2). These results suggest a successful engineering of a deoxysugar pathway to generate novel hybrid macrolide derivatives, including deoxyaminosugar.     

Assessment of protein spot components applying correspondence analysis for peptide mass fingerprint data

Proteins separated by two-dimensional gel electrophoresis (2-DE) may be distributed over several spots. Otherwise, one spot may contain more than one component. The same protein occurring in several spots supposedly represents differently modified protein species that might be of biological relevance. Identification of spots with peptide mass fingerprinting and database searching leads only to the detection of the major spot components. If a spot also contains additional minor protein components, quantitation of spots with protein staining techniques or antibody detection becomes misleading. In order to find spots containing minor components we applied correspondence analysis, a multivariate data exploration method, to peptide mass fingerprint data. Correspondence analysis using peak lists revealed groups of spots containing the same protein with their characteristic mass-to-charge ratio (m/z) values. In order to detect different protein spot components an interactive threshold setting and removal of m/z values with subsequent recalculation of the correspondence analysis using our software tool CorrAn are performed. The usefulness of this methodical approach was shown by a data set of peptide mass fingerprints of 284 spots of Helicobacter pylori 26695 separated by 2-DE.     

Metabolite profiling of plant carotenoids using the matrix-assisted laser desorption ionization time-of-flight mass spectrometry

Although modern MS has facilitated the advent of metabolomics, some natural products such as carotenoids are not readily compatible to detection by MS. In the present article, we describe how matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI/TOF-MS) can be utilized to acquire mass spectra of carotenoids effectively. The procedure is sensitive (pmole range), reduces 'spot to spot' variation and provides high mass accuracy, thus aiding identification. The technique has been applied in vivo to the analysis of carotenoids in isolated plant cells and in vitro to three applications: (i) to show compatibility with purification methods such as LC, TLC and HPLC; (ii) for the rapid identification and quantification (by isotope dilution) of carotenoids present in crude extracts from plant tissues and whole cells; (iii) simultaneous semi-quantitative determination of carotenoids metabolites (m/z values) in crude plant extracts. Multivariate analysis of the recorded m/z values shows the effectiveness of the procedure in distinguishing genotypes from each other. In addition, the utility of the technique has been demonstrated on two mutant tomato populations, to determine alterations in carotenoid content, and a comparison made with traditional HPLC-photodiode array analysis. These data show that MALDI/TOF-MS can be used to rapidly profile, identify and quantify plant carotenoids reproducibly, as well as detecting other metabolites (m/z) in complex biological systems.     

Intact glucosinolate analysis in plant extracts by programmed cone voltage electrospray LC/MS: performance and comparison with LC/MS/MS methods

We present a comprehensive, sensitive, and highly specific negative ion electrospray LC/MS method for identifying all structural classes of glucosinolates in crude plant extracts. The technique is based on the observation of simultaneous maxima in the abundances of the m/z 96 and 97 ions, generated by programmed cone voltage fragmentation, in the mass chromatogram. The abundance ratios lie in the range 1:2-1:4 ([m/z 96]/[m/z 97]). Examination of the corresponding full-scan mass spectra allows individual glucosinolates of all structural classes to be identified rapidly and with confidence. The use of linearly programmed cone voltage fragmentation enhances characteristic fragment ions without compromising the abundance of the analytically important [M - H]- ion and its associated (and analytically useful) sulfur isotope peaks. Detection limits are in the low nanogram range for full-scan, programmed cone voltage spectra. Comparison of the technique with LC/MS/MS methods (product ion, precursor ion, and constant neutral loss scans) has shown that the sensitivity and selectivity of the programmed cone voltage method is superior. Data obtained on a variety of plant extracts confirmed that the methodology was robust and reliable.     

Determination of Simvastatin in human plasma by liquid chromatography-mass spectrometry

A simple, sensitive and selective liquid chromatography coupled with electrospray ionization mass spectrometry (LC/ESI/MS) method for the determination of simvastatin (I) has been developed. After extraction by ethyl acetate, using lovastatin (II) as internal standard, solutes are separated on a C(18) column with a mobile phase consisting of methanol-water (9:1). Detection is performed on an atmospheric pressure ionization single quadruple mass spectrometer equipped with an ESI interface and operates in positive ionization mode. Simvastatin quantification was realized by computing peak area ratio (I/II) of the extracts analyzed in SIM mode (m/z: 441 and m/z: 427 for I and II, respectively) and comparing them with calibration curve (r=0.9997). Accuracy and precision for the assay were determined by calculating the intra-batch and inter-batch variation at three concentrations 0.1, 5.0, 10.0 ng/ml; the intra batch relative standard deviation (RSD) was less than 10% and ranged from 1.8 to 8.5%, respectively; the inter-batch RSD was less than 20% and ranged from 4.1 to 16.5%. The limit of detection was 0.05 ng/ml.     

Evaluation of the Ecotoxicity of Sediments from Yangtze River Estuary and Contribution of Priority PAHs to Ah Receptor-Mediated Activities

In this study, in vitro bioassays were performed to assess the ecotoxicological potential of sediments from Yangtze River estuary. The cytotoxicity and aryl hydrocarbon receptor (AhR)-mediated toxicity of sediment extracts with rainbow trout (Oncorhynchus mykiss) liver cells were determined by neutral red retention and 7-ethoxyresorufin-O-deethylase assays. The cytotoxicity and AhR-mediated activity of sediments from the Yangtze River estuary ranged from low level to moderate level compared with the ecotoxicity of sediments from other river systems. However, Yangtze River releases approximately 14 times greater water discharge compared with Rhine, a major river in Europe. Thus, the absolute pollution mass transfer of Yangtze River may be detrimental to the environmental quality of estuary and East China Sea. Effect-directed analysis was applied to identify substances causing high dioxin-like activities. To identify unknown substances contributing to dioxin-like potencies of whole extracts, we fractionated crude extracts by open column chromatography. Non-polar paraffinic components (F1), weakly and moderately polar components (F2), and highly polar substances (F3) were separated from each crude extract of sediments. F2 showed the highest dioxin-like activities. Based on the results of mass balance calculation of chemical toxic equivalent concentrations (TEQs), our conclusion is that priority polycyclic aromatic hydrocarbons indicated a low portion of bio-TEQs ranging from 1% to 10% of crude extracts. Further studies should be conducted to identify unknown pollutants.     

Analysis of polar lipids from some representative enterobacteria, Plesiomonas and Acinetobacter by fast atom bombardment-mass spectrometry.

Fast atom bombardment-mass spectrometry (FAB-MS) was used to analyse lipid extracts of bacteria to assess its usefulness for analysing anionic phospholipids of potential chemotaxonomic value. The following micro-organisms were tested: Acinetobacter calcoaceticus, Acinetobacter sp., Citrobacter freundii, Enterobacter cloacae (2 strains), Escherichia coli (3 strains), Hafnia alvei, Klebsiella oxytoca, Klebsiella pneumoniae, Morganella morganii, Plesiomonas shigelloides, Proteus mirabilis (3 strains), Serratia liquefaciens and Serratia marcescens. Negative-ion spectra provide data for twenty-seven major carboxylate anions (m/z 209-325) and for thirty-seven major phospholipid anions (m/z 645-774). Generally, the largest carboxylate peaks were due to 16:1, 16:0, cyc17 and 18:1 while the largest phospholipid anion peaks were due to PE(32:1), PE(33:1), PE(34:1), PE(34:2), PG(30:2), PG(31:2), PG(32:2), PG(34:1) and PS(33:0). However, quantitative differences were observed. For example, Acinetobacter lacked PE (33:1) but had exceptionally high peaks at m/z 748, PS(33:0), and m/z 281, octadecanoate. Unknown 'carboxylate' peaks were detected at m/z 254, 256, 261, 268, 282 and 301. In some cases, unknown peaks appeared to constitute possible homologous series being separated by delta m/z of 14(identical to methylene). For chemotaxonomic purposes, the complexity of the data required numerical analysis. Using the Pearson coefficient of linear correlation, as a measure of association, it was possible to compare all strains analysed. Typical results for strain comparisons were as follows: Ent. cloacae vs Ent. cloacae, r = 0.90 (Ent. cloacae vs Ac. calcoaceticus, r = 0.46). Thus FAB-MS represents an excellent means of obtaining large quantities of data on polar lipids of a range of bacterial isolates, which may be suitable for chemotaxonomic purposes.     

Analysis of polar lipids from some representative enterobacteria, Plesiomonas and Acinetobacter by fast atom bombardment-mass spectrometry

Fast atom bombardment-mass spectrometry (FAB-MS) was used to analyse lipid extracts of bacteria to assess its usefulness for analysing anionic phospholipids of potential chemotaxonomic value. The following micro-organisms were tested: Acinetobacter calcoaceticus, Acinetobacter sp., Citrobacter freundii, Enterobacter cloacae (2 strains), Escherichia coli (3 strains), Hafnia alvei, Klebsiella oxytoca, Klebsiella pneumoniae, Morganella morganii, Plesiomonas shigelloides, Proteus mirabilis (3 strains), Serratia liquefaciens and Serratia marcescens. Negative-ion spectra provide data for twenty-seven major carboxylate anions (m/z 209–325) and for thirty-seven major phospholipid anions (m/z 645–774). Generally, the largest carboxylate peaks were due to 16: 1, 16: 0, cyc17 and 18: 1 while the largest phospholipid anion peaks were due to PE(32: 1), PE(33: 1), PE(34: 1), PE(34: 2), PG(30: 2), PG(31: 2), PG(32: 2), PG(34: 1) and PS (33: 0). However, quantitative differences were observed. For example, Acinetobacter lacked PE (33: 1) but had exceptionally high peaks at m/z 748, PS(33: 0), and m/z 281, octadecanoate. Unknown 'carboxylate' peaks were detected at m/z 254, 256, 261, 268, 282 and 301. In some cases, unknown peaks appeared to constitute possible homologous series being separated by Δ m/z of 14(≡ methylene). For chemotaxonomic purposes, the complexity of the data required numerical analysis. Using the Pearson coefficient of linear correlation, as a measure of association, it was possible to compare all strains analysed. Typical results for strain comparisons were as follows: Ent. cloacae vs Ent. cloacae, r = 0.90 ( Ent. cloacae vs Ac. calcoaceticus, r = 0.46). Thus FAB-MS represents an excellent means of obtaining large quantities of data on polar lipids of a range of bacterial isolates, which may be suitable for chemotaxonomic purposes.