灵芝TR6号漆酶的分离纯化及性质研究

采用硫酸铵盐析、DEAE Sepharose Fast Flow柱层析和Sephadex G100凝胶柱层析对灵芝TR6号漆酶发酵液进行分离纯化,得到电泳纯漆酶.SDS-PAGE电泳结果显示,该漆酶表观分子量为62×103.Native-PAGE鉴定该漆酶由一种同工酶组成,为单体酶.试验结果表明,该漆酶的最适反应温度为60℃,与ABTS的最适反应pH为4.5,Km为0.188 mmol/L,在pH 6.0～8.0之间较稳定.Fe2 、SDS和Tris对该漆酶活性具有抑制作用,EDTA和Tween80则对该漆酶活性具有促进作用.图8表2参10     

灵芝深层发酵生产胞外多糖和灵芝酸的动力学分析

利用Sigmoidi函数构建了灵芝深层发酵生产胞外多精和灵芝酸的非结构动力学模型,并根据Boltzmann拟合求解出模型参数,模型预测值能够较好地吻合实验所测值.细胞最大比生长速率цmax为4.63x10-2 h-1,葡萄糖最大比消耗速率qs,max为6.70x10-2 h-1,胞外多糖最大比合成速率qEPs,max为4.65x10-3h-1,灵芝酸最大比合成速率qGA,max为9.09xlO-4 h-1.灵芝胞外多糖的合成与细胞的生长呈现部分偶联关系;灵芝酸的合成与细胞的生长呈现偶联关系,偶联系数αGA为0.020 4 g g-1.胞外多糖对葡绚糖的最大得率系数(YEPS/S)为O.214 g g-1;灵芝发酵40～80 h代谢碳流迅速从菌体自身生长迁移至胞外多糖合成,用于合成胞外多糖的最大碳流为23.60/0.     

灵芝中重金属的检测及其健康风险初步评价

为了研究灵芝中重金属的污染状况,对北京市一些药店及部分中医门诊部所售的不同产地灵芝中的As、Hg、Pb等重金属元素含量进行了测定.结果表明,灵芝中As含量范围为0.016￣0.239mg·kg-1,平均值0.117mg·kg-1,Hg含量范围从未检出到0.43mg·kg-1,平均值0.115mg·kg-1,Pb含量范围从未检出到0.256mg·kg-1,平均值0.047mg·kg-1,As、Pb含量均符合我国《药用植物及制剂进出口绿色行业标准》,Hg有5例超标,占样品总数的25%,主要是野生灵芝.健康风险初步评价结果表明,服用灵芝的人群,成人每人每日通过灵芝摄入As、Hg、Pb分别为0.18￣2.3μg、0.17￣2.3μg、0.07￣0.94μg,分别占每日允许摄入量(ADI)的0.14%￣1.9%、0.4%￣5.4%、0.03%￣0.4%,对人体健康风险不大.但是对于个别野生和人工种植灵芝而言,每日摄入总汞量可达0.47￣6.24μg,占ADI的1.1%￣15%,对人体健康存在一定的风险.     

女贞枝叶挥发物对小白鼠自发活动的影响

采用旷场实验,通过连续6天的实验观察,分析昆明种小白鼠在不同浓度女贞枝叶挥发气味环境中处理后其行为指标的变化,研究了女贞枝叶挥发物对小白鼠自发活动的影响,以此来客观评价女贞挥发物的保健功能。结果表明:1)处理组小白鼠运动总路程大于对照组,表明女贞挥发物有助于提高小白鼠的兴奋性。2)中央活动路程和站立次数两项指标呈现阶段性变化,探究适应阶段处理组小白鼠的指标数值显著高于对照组,熟悉环境阶段组间差异减小,但处理组的指标数值下降幅度小于对照组,表明女贞挥发物有利于促进小白鼠的探究性行为,提高其感知能力、学习和记忆力水平。3)实验过程中处理组小白鼠的粪便粒数始终比对照组少,表明女贞枝叶挥发物有利于缓解小鼠对新环境的紧张状态。     

Discharge characteristics of nematocysts isolated from acontia ofCalliactis parasitica

Experiments were performed on specimens ofCalliactis parasitica collected near Plymouth in April 1987 and in the Mediterranean between September 1987 and April 1988. Undischarged nematocysts (basitrichous isorhizas) were separated from the acontia of by using 1M glycerol, 1M citrate, 0.5% Nonidet or 0.5% Triton X-100 as isolating agents, or by freezing the acontial tissue. The mechanisms of extrusion were not studied. The effectiveness of 50 mM thioglycolate in discharging nematocysts isolated by the above methods and suspended in Ca- and Mg-free artificial sea water (ASW) was investigated. Nematocysts extruded in glycerol were more responsive to thioglycolate than those extruded in citrate. Capsules isolated in non-ionic detergents, however, and those obtained by freezing were not discharged by thioglycolate. If capsules extruded in glycerol were suspended in ASW and this was replaced with distilled water, they did not respond to thioglycolate. The effect of distilled water was only partly reversible. It was observed that these nematocysts, unlike those ofPelagia noctiluca andAiptasia mutabilis previously investigated, did not release measurable amounts of free Ca²⁺ during discharge. It is suggested that if calcium is not involved in the mechanism of discharge of isolatedC. parasitica nematocysts then some other ionic species may stabilize the resting condition of these capsules.     

生物与化学表面活性剂对多氯联苯的协同增溶作用

采用室内序批试验比较研究了单一的生物/非离子化学表面活性剂(SAA)与生物-非离子混合表面活性剂对商业用多氯联苯(PCBs)Aroclor1242的增溶作用.结果表明,生物表面活性剂鼠李糖脂(RL)对Aroclor1242的增溶作用要高于三种非离子SAA,三种非离子SAA对Aroclor1242的增溶作用顺序为POE(6)＞POE(10)＞Brij35,与其亲水亲油平衡值(HLB)呈负相关;鼠李糖脂与非离子SAA对Aroclor1242的增溶存在协同效应,混合表面活性剂溶液中Aroclor1242的摩尔增溶比(MSR)、胶束相/水相间的分配系数(Kmc)均大于对应的单一表面活性剂,其协同增溶作用顺序为RL-Brij35＞RL-POE(10)＞RL-POE(6),协同增溶作用的大小与其中的非离子表面活性剂的HLB值呈正相关.     

New denitrifying bacteria isolated from Red Sea sediments

Sediments of various newly discovered deeps in the Red Sea were analyzed for the occurrence of denitrifying bacteria. The samples were collected in 1972 during the Valdivia cruises. Among the 27 different samples investigated, 17 revealed both coccoid and rod-shaped bacteria when enriched in complex nutrient broth (with 10% NaCl). Denitrifiers were recorded abundantly in the sediments, their population decreasing from some 10⁶/g in the surface material to only a few in the subsediment. A total of 16 pure cultures of denitrifying bacteria were isolated from the Suakin-and Thetis-deeps and studied morphologically, physiologically and biochemically. Genetics (molar percentage of guanine plus cytosine, % GC) and numerical taxonomy were included to reveal relationships and improve taxonomic classification. Fifteen isolates were described as Gram-negative, aerobic and facultative anaerobic (with NO ₃ ^- as H⁺-acceptor), polarly flagellated rods (Pseudomonas spp.); one was an inmotile, Gram-positive, facultative anaerobic coccus. None of the 15 Pseudomonas-isolates could be identified with one of the denitrifying species so far described and recognized. The strains should be regarded as hitherto undescribed denitrifying marine bacteria.Supported by the Bundesministerium für Forschung und Technologie, Bonn, Germany (FRG).     

Low-temperature adaptation of deep-sea bacteria isolated from the Japan Trench

Montastraea annularis, M. faveolata, and M.␣franksi are three recently separated species that together dominate reefs of the tropical western Atlantic. Despite morphological, life-history, ecological and genetic differences, the legitimacy of their status as separate species has been questioned. This controversy stems from both the scarcity of unambiguous, diagnostic differences among them, and from the possibility of extensive hybridization associated with their approximately synchronous reproduction in sympatry. Here we report on fertilization trials and the timing of spawning, both of which suggest that the potential for hybridization may be limited. Crosses between M. faveolata (the most genetically distinctive taxon) and the other two species were largely unsuccessful, as were selfed matings for all three species. M. annularis and M. franksi showed no evidence of fertilization barriers, but the timing of spawning typically differed between them by 1 to 2 h. We also found that spawning times in the field of M. annularis and M. faveolata were non-overlapping in 1995, and that the timing of spawning in M. annularis could be experimentally shifted forward by simulating earlier than natural sunsets. These findings from Panamá and Honduras, particularly given their consistency with comparable observations and experiments elsewhere, provide evidence of reproductive isolation and support the separate species status of these three taxa. Some hybridization may occur under natural conditions at these sites, particularly between M. annularis and M.␣franksi. Rampant genetic interchange among these taxa in Central America seems unlikely, however, in the light of the concordant associations between morphology, reproductive biology and other characters which are observed. Received: 22 July 1996 / Accepted: 2 October 1996     

Characterisation and identification of carbofuran-utilising bacteria isolated from agricultural soil

A total of 96 bacterial cultures were isolated from soil. Seventeen bacterial isolates were selected following their cultivation on solid media containing 100 mg · L^?1 carbofuran as the sole source of carbon and nitrogen. Of the 17 isolates, 10F, 11M, 17N, 23B and 26M were specifically chosen because of their relatively higher growth efficiency and genetic diversity based on Box-polymerase chain reaction analysis. These bacterial cultures had 16S rRNA gene sequences that were most similar to Acinetobacter baumannii (10F), Agrobacterium tumefaciens (11M), Ochrobactrum anthropi (17N), Escherichia coli (23B) and Agrobacterium tumefaciens (26M) with 97, 95, 93, 95 and 94% similarity in their 16S rDNA gene sequence, respectively. Degradation rates of carbofuran in soil inoculated with these isolates were 1.9, 1.5, 1.6, 1.7 and 1.6 times, respectively, faster in comparison with uninoculated soil after 10 days of incubation. The maximum degradation rates of carbofuran (45 and 91%) were detected in soil inoculated with A. baumannii (10F) after 10 and 20 days’ incubation, respectively. These data indicate that these isolates may have the potential for use in bioremediation of pesticide contaminated soil.     

不同类型表面活性剂对1，2，4-三氯苯的增溶作用

研究了阴离子表面活性剂十二烷基硫酸钠、阳离子表面活性剂十六烷基三甲基溴化铵和非离子表面活性剂脂肪醇聚氧乙烯醚LAE对1,2,4-三氯苯（TCB）的增溶作用。讨论了表面活性剂类型对增溶作用的影响。比较发现,非离子型表面活性剂的增溶能力高于离子型表面活性剂。其原因主要是疏水有机物在非离子表面活性剂胶束中的增溶方式不同造成的。3种表面活性剂对TCB的增溶数据还表明,增港作用不仅在临界胶束浓度（MC）以上发生,而且在表面活性剂浓度低于CMC时亦有明显的增溶作用。     

Effect on fishes of two toxic polypeptides isolated from Anemonia sulcata

The effects of two toxic polypeptides isolated from Anemonia sulcata Pennant (Anthozoa) were tested on Gadus morhua, Pleuronectes platess and Platichthys flesus. In tramuscular injections caused paralysis and death, with symptoms similar to those of death caused by lack of oxygen. In cod, the LD₅₀ dose of Toxin II was 50 times higher chan that of Toxin I (Toxin I=6.9 mg/kg, Toxin II=0.14 mg/kg). Flatfish were less sensitive. The LD₅₀ of Toxin II was 0.37 mg/kg for Platichthys flesus and 0.38 mg/kg for Pleuronectes platessa. Both toxins are less lethal to fishes than to crustaceans.     

Effect of neem extract against the bacteria isolated from marine fish

Marine ornamental fishes are exceedingly valuable due to their high demand in domestic and international markets. There is a growing global interest to rear the fishes in captivity. But problem due to bacteria and fungi are the major hitch in captive condition. Since, the use of antibiotics is banned, an attempt was made to ascertain in vitro assay of the neem leaves extract against the bacterial pathogens isolated from infected fishes. Bacterial strains isolated from infected regions of the clown fishes Amphiprion sebae and A. ocellaris were identified as Aeromonas hydrophila, Enterobacter sp., E. coli, Pseudomonas aeruginosa, Proteus sp., Streptococcus sp., Vibrio cholerae, V. alginolyticus, V. parahaemolyticus and Yersinia enterocolitica. Ethanol and methanol extracts were highly inhibitory to the bacterial isolates when compared to other solvents. Ethanol extracts exhibited low minimum inhibitory concentration (75-250 microg ml(-1)) as compared to other extracts. The present finding revealed that the neem leaf extract significantly reduces the bacterial pathogens and their infection in marine ornamental fishes.     

Biodegradation of phenol by native microorganisms isolated from coke processing wastewater

The present investigation was undertaken to assess the biodegradation of phenol by native bacteria strains isolated from coke oven processing wastewater. The strains were designated ESDSPB1, ESDSPB2 and ESDSPB3 and examined for colony morphology Gram stain characters and biochemical tests. Phenol degrading performance of all the strains was evaluated initially. One of the strains namely ESDSPB2 was found to be highly effective for the removal of phenol, which was used as sole carbon and energy source. From an initial concentration of 200 mg I(-1) it degraded to 79.84 +/- 1.23 mg l(-1). In turn the effect of temperature (20 to 45 degrees C), pH (5-10) and glucose concentration (0, 0.25 and 0.5%) on the rate of phenol degradation by that particular strain was investigated. Observations revealed that the rate of phenol biodegradation was significantly affected by pH, temperature of incubation and glucose concentration. The optimal conditions for phenol removal were found to be pH of 7 (84.63% removal), temperature, 30 degrees C (76.69% removal) and 0.25% supplemented glucose level (97.88% removal). The main significance of the study is the utilization of native bacterial strains from the waste water itself having potential of bioremediation.     

Sulphur-oxidising bacteria isolated from deep caves improve the removal of arsenic from contaminated harbour sediments

Acidophilic S-oxidising bacteria isolated from sulphur-rich deep caves (Frasassi, Italy), characterised by relatively low temperature, were tested for their ability to mobilise (semi-)metals from contaminated sediments. Sediment samples from two commercial Italian seaports were used to set up bioleaching experiments. The effect of different growth substrates was also investigated. Our experiments revealed that S-oxidising bacteria isolated from Frasassi caves have a high potential to remove As from contaminated marine sediments, as never reported before. Although As solubilisation efficiency was quite low (i.e. about 30%), only a small amount of As was associated with non-residual fractions of the sediment. On the contrary, the solubilisation efficiencies of Zn and Ni (20% and 10%, respectively) were lower than those previously obtained by the use of other acidophilic bacteria and mainly influenced by the experimental conditions rather than by the presence of the S-oxidising bacteria. Results presented here open new perspectives in bioleaching applications for the remediation of contaminated sediments. Indeed, microbial strains adapted to relatively low-temperature environments could improve sediment bioleaching at temperature regimes where mesophilic and thermophilic strains are not favoured. Such strains could be exploited for developing selective bioremediation procedures for sediments contaminated with As, to be applied in multistep biotreatment processes.     

Abundant ketone isolated from oily Plakortis sponge demonstrates antifouling properties

Marine sponges of the Genus Plakortis are typically unfouled; they can have a distinctive pleasant smell and an oily surface. A significant quantity of fragrant oil was obtained from a Jamaican Plakortis sp. by cryo-trap. The oil was determined to be exclusively 2-decanone. The antifouling character of the oil was evaluated by its effects on surface attachment of a gram negative bacterial model using confocal fluorescence microscopy as well as its effects on the attachment of Dreissena polymorpha (zebra mussel). The ketone dramatically inhibited attachment of the bacteria and zebra mussels. The results suggest that the oil impacts establishment of related epifauna on the Plakortis sponge in nature. Although the aliphatic ketone alone is not a potential commercial alternative for antifouling coatings, incorporating the functionality into coating design should be considered in future studies.     

Biodegradation of Hexachlorocyclohexane by two species of Bacillus isolated from contaminated soil

Biodegradation of α, β, γ and δ hexachlorocyclohexane (HCH) isomers was studied in broth medium and soil microcosm by Bacillus circulans and Bacillus brevis isolated from contaminated soil. Degradation of α and γ isomers by both the bacterial isolates was higher than thermodynamically stable β and δ isomers. However, B. circulans was found more effective than B. brevis for β and δ isomers. Maximum rate of degradation was recorded at 150 mg/L followed by 100 and 50 mg/L. Soil microcosm study revealed maximum degradation of HCH isomers in the treatment containing natural soil, pesticide and bacterial inocula than the treatment having sterilised soil, pesticide and bacterial isolates. Chloride release was positively co-related with HCH degradation in broth medium as well as in soil microcosm, suggesting that B. circulans and B. brevis hold promising potential by having efficient enzyme(s) required for dechlorination of HCH from contaminated sites.     

鹅掌楸和女贞同化CO2和释O2能力的比较

采用LI－6400红外气体分析仪，分别在生长初期、盛期、末期测定鹅掌楸和女贞净光合速率，用重量法测定叶面积。得到女贞的净光合速率、叶面积指数和同化CO2及释O2的能力均大于鹅掌楸的结论，可为城市绿化树种的选择提供依据。     

Bacteriolytic Bacillus species isolated from brackish waters of the Southern Baltic Sea

The present study examines antagonistic relationships between different microorganisms inhabiting brackish water and includes a systematic screening for bacteriolytic prokaryotes in the brackish waters of the Southern Baltic Sea (Nordruegenscher Bodden) sampled in July/August 2009. Ten of the 35 marine bacteriolytic isolates belong to the genus Bacillus. Five isolates (B. pumilus, B. subtilis, B. megaterium, B. licheniformis) lysed living microbial cells such as the bacteria Arthrobacter citreus, Arthrobacter protophormiae, Micrococcus luteus, Pseudomonas putida and the yeast Trichosporon mucoides. These and other bacteria (Aeromonas sp., Bacillus subtilis, Chromobacterium violaceum, Citrobacter freundii, Enterobacter aerogenes, Proteus mirabilis, Pseudomonas aeruginosa and Serratia marcescens) were also lysed as autoclaved and pasteurized cells on agar plates. One isolate of Bacillus pumilus showed a distinct bacteriolysis activity against pasteurized cells of A. citreus in liquid culture. Our results suggest that Bacillus species may play a role as opportunistic predators in the marine microbial food web.     

Biodegradation of paclobutrazol by a microbial consortium isolated from industrially contaminated sediment

Biodegradability of the plant growth retardant paclobutrazol by a microbial consortium in which Pseudomonas was the predominant strain was investigated in batch culture. The consortium which had been isolated from an industrially contaminated sediment was proven to be useful for the treatment of effluents containing paclobutrazol. Paclobutrazol was degraded by the pure isolated strain of Pseudomonas sp. as well as the microbial consortium. Paclobutrazol was utilized as the sole source of carbon and energy. Sixty percent of the paclobutrazol was degraded by the microbial consortium from an initial concentration of 54 mg L^?1 within 48 h and more than 98% of an initial concentration of 3.4 mg L^?1 was degraded within 36 h. The optimum temperature and pH were determined to be 30°C and 7.0, respectively. A pure strain of a bacterium, isolated from the enrichment culture was identified as Pseudomonas sp. The microbial consortium was tolerant of high pH and could degrade paclobutrazol faster than the pure strain. The degradation rate of this plant growth regulator in an aerobic environment was greater than that under anaerobic conditions.     

Influence of 1,2,4,5-tetrazine derivatives on growth of bacterial consortium isolated from soil

This article presents the results of laboratory studies of the influence of tetrazine derivatives on the growth kinetic parameters of soil bacteria. 3,6-Dihydrazinotetrazine (DHTz), 3,6- bis(3,5-dimethylpyrazol-1-yl)-dihydro-1,2,4,5-tetrazine (DMPDHT) and N,N′-bis(1,2,4,5-tetrazine-6-(3,5-dimethylpirazylo))hydrazine (BDMPT) were applied. 3,6-Dihydrazinetetrazine had the largest influence on the growth of bacteria, reflected in a significant lengthening of the lag-phase and a decrease in the specific growth rate. Dehydrogenase activity was also determined in bacterial cultures exposed to tetrazine derivatives. Dehydrogenases remained active even at DHTz concentrations of 80 mg · L^?1, which completely inhibited bacterial growth. The compounds studied variously influence the kinetics of growth in the bacterial consortium; at the same time, they undergo biodegradation in soil by autochthonous microflora.