Two phenol-degrading bacterial strains named XTT-1,XTT-3 were isolated from active sludge using the way of acclimatization and could grow on a medium with phenol as the sole carbon and energy sources,the two strains were identified preliminarily as Pseudomonas sp.The optimum degradation condition of the two strains was explored.Strain XTT-3 could degrade phenol completely at a concentration as high as 500 mg·L-1 in 48 h,while strain XTT-1 did it in 64h..The two strains could resist to phenol up to 1000 mg·L-1.Salt content higher than 2.0% had different degrees of inhibition on phenol degradation.

OBJECTIVE:The study was to provided the reference for further development and utilization of Ligularia hodgsonii.METHODS:The crude polysaccharide LW was extracted from Lugularia hodgsonii with hot water, and prectated by ethanol. The crude polysaccharide has been fractionted and purified, so LW21 was got. Its homogeneity was confirmed by chromatography on Sepharose CL-4B, paper chromatography and cellulous actate electrophoresis. The scavenging effets of the polysacehardes on * OH and O2-* were determined by fluorescence method.RESULTS:Antioxidative experiment shows that LW and LW21 were effective in scavenging superoxide radical and hydroxyl radical. The scavenging capacity for superoxide radical and hydroxyl radical of LW21 had higher than that of LW.CONCLUSION:The polysaceharide from Ligularia hodgsonii had stronger antioxidant actioxdant activities, and their antioxidant activities had certain dose-relationship.

Chloroperoxidase(CPO)from Caldariomyces fumago is a highly versatile oxidase.But its application is limited,because of its low accumulated concentration in the biological synthesis,poor dynamic stability,low recovery of high-purity enzyme in purification process and high price.A novel very efficient method has been developed.At first,CPO was precipitated with hydrophilic polymer(PEG 6000)in the high-saturated solution of aluminum sulfate.And then it was extracted by an aqueous two-phase system of PEG/ammonium sulfate.At last,the extracted CPO was purified by Sephadex G-75 chromatographic system.The results showed that:60% saturated(NH4)2SO4 solution could make PEG 6000 precipitate 95% chloroperoxidase.In the aqueous two-phase system CPO was prior to dissolve in bottom phase,its partition coefficient k was lower than 0.228,its yield about 65.2%,its purification was 7.4 times.Finally,the purity of CPO was slightly higher after chromatographic fractionation.The method was also provide a new method for extracting efficiently other trace biological molecules.

The hydrophilic polymer PEG6000 was used to co-precipitate with chloroperoxidase(CPO)in the saturated ammonium sulfate solution.Then,a aqueous two-phase system of PEG/phosphate was formed based on the co-precipitation resolution to extracted higher composition CPO,and the purity CPO sample from Sephadex G-100 chromatographic system was obtained.Results showed that the CPO yield was 85.5% by co-precipitate with PEG in the presence of the phosphate solution.In the aqueous two-phase system,CPO was dissolved in the bottom phase,the partition coefficient k was lower than 0.341,the yield reached 69.1%,and the purification ratio increased 21.57 times.The purity of CPO was further improved to 24.79 times by column chromatography,the total yield of CPO was 37.75%.

The crude polysaccharide was extracted from cultivate Taxus with hot water,and precipitated by ethanol.The rusults thawed that it was no starch in the crude polysaccharide,The contents of protein was 7.0%and cultivate Taxus polysaccharide(CTP) was 19.71%.The scavenging capacity for superoxide radical and hydroxyl radical of refined CTP was higher than that of crude CTP.

为合理利用雪莲提供一定依据,对新疆天然雪莲进行多糖提取分离纯化.新疆天然雪莲经热水浸提,乙醇沉淀得粗多糖.粗多糖经Sevage法和酶法联合脱蛋白,酸性乙醇分级,得初步纯化的多糖XL1,XL2,XL3.其中XL3经DEAE-SephdexA25分离纯化得纯化多糖XL31.XL31经检测为纯多糖.气相色谱分析表明新疆天然雪莲多糖XL1,XL2,XL3和XL31均为Ara、Rha、Xyl、Gal、Glc、GalA六种单糖组成的酸性杂多糖,但单糖的摩尔比不同.

通过采用热水提取乙醇沉淀获得雪莲水溶性粗多糖,经酸性乙醇分级和DEAE-SephdexA-25纯化得多糖XL31.纸层析、醋酸纤维薄膜电泳和Sepharose CL-4B柱层析纯度鉴定表明XL31为均一多糖,分子量约为170kD.其单糖组成为阿拉伯糖(Ara)、鼠李糖(Rha)、木糖(Xyl)、半乳糖(Gal)、葡萄糖(Glc)、半乳糖醛酸(GalA),摩尔比为11:4:1:18:3:9.XL31结构分析采用了高碘酸氧化、Smith降解、甲基化分析及IR、NMR、GC和GC-MS等方法.结果表明:多糖XL31主链由Ara、Gal、GalA构成,其中Ara主要以β-(1→4)或β-(1→5)糖苷键连接,在3-O处有分枝,Gal主要以β-(1→6)及β-(1→4)糖苷键连接,β-(1→6)糖苷键连接在3-O和4-O处有分枝,β-(1→4)糖苷键连接在2-O、3-O和6-O处有分枝;GalA以α-(1→4)糖苷键连接.支链由Xyl、Rha、Glc构成,其中Xyl以1→4或1→5糖苷键连接;Rha以1→2、4糖苷键连接;Glc以1→4糖苷键连接.末端残基为GalA、Gal、Ara、Xyl、Rha、Glc.雪莲多糖XL31是一新结构多糖,为首次从新疆雪莲中分离得到.

In this experiment,the polysaccharide was extracted and purified from Toona sinensis Roem.We measured and purified the number of the polysaccharide protein,made an anti-oxide experiment for water-soluble polysaccharide and the purified polysaccharide,and measured the effect of eliminate of free radical for the different density of polysaccharide.The result shows that the composition of the polysaccharide was Xyl,Ara,Rha,Gal,Glc,Gal-A,and Man,and proportion of mol:1.0 ∶2.8 ∶2.6 ∶2.6 ∶2.0 ∶7.6 ∶1.6.A result obtained from anti-oxide experiment shows that different density of water-soluble polysaccharide and purified polysaccharide can eliminate free radical·OH and anions.Furthermore,the effect of purified polysaccharide is beyond of water-soluble polysaccharide.The water-soluble polysaccharide can eliminate R and the purified polysaccharide can not eliminate it.

采用热水浸提醇沉法从新疆雪莲中提取水溶性粗多糖XL。此粗多糖经分离纯化得多糖XL31。XL31经检测为纯多糖。对新疆雪莲多糖XL31进行清除自由基活性及抑菌活性研究,结果表明XL31对羟自由基及超氧阴离子自由基具有明显的清除效果,但对脂质自由基的清除效果不明显;XL31对细菌有较强的抑制作用,对真菌无抑制作用。

To optimize the quality and quantity of polysaccharides from Saussurea involucrata,a water-soluble polysaccharide was extracted from Saussurea involucrata by method of boiling water and ethanol deposition.Reagent proportion,reaction temperature and reaction time were selected with substitution degree and yield as index by L9(33) orthogonal design.The result shows that the optimal extraction conditions were 85 ℃,2 h as well as the ratio 1∶12 v/v.By Combination with the optinums condition,a high crude polysaccharides(7.08%) was obtained,and the content of polysaccharide was 15.37%.

The crude polysaccharide was extracted from Ligularia duciformis with hot water,and precipitated by ethanol.Then some identifiable properties of the crude polysaccharide were studied. The contents of protein was 14.02%,the contents of polysaccharide was 23.41% and the contents of GAL-A was 35.16%.The refined polysaccharide was prepared after protein being separated.Both crude and refined polysaccharide were effective in scavenging superoxide radical and hydroxyl radical.The scavenging capacity for superoxide radical and hydroxyl radical of refined polysaccharide was higher than that of crude polysaccharide.

The crude polysaccharide was extracted from Tibetan Saussurea with hot water,and precipitated by ethanol.Then some identifiable properties of the crude polysaccharide were studied.It contained of no starch,The contents of protein was 4.38%,the contents of Tibetan Saussure polysaccharide(TSP) was 25.8% on phenolvitriol method and the contents of GAL-A was 38.18%.The refined polysaccharide was prepared after protein being separated.Based on PC analysisand GC analysis,it showed that TSP was composed of seven different kinds of sugar :Xyl,Ara,Rha,Gal,Glc,Gal-A,Man,proportion of mol:1.0∶2.8∶2.6∶2.6∶2.0∶7.6∶1.6.Both crude and refined TSP were effective in scavenging superoxide radical and hydroxyl radical.The scavenging capacity for superoxide radical and hydroxyl radical of refined TSP had higher than that of crude TSP.Furthermore,TSP had only a little effect in scavenging lipid radical.