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1.
黑柄炭角菌产生的DPPH自由基捕捉成分 总被引:10,自引:0,他引:10
对黑柄炭角菌深层发酵制品中的DPPH自由基捕捉成分进行研究。经硅胶柱层析、中压液相色谱顺相和反相分离、制备型高压液相色谱分离等一系列步骤 ,共获得相对纯度在85%以上 ,收量在 2mg以上的自由基捕捉物质 2 0个 ,对其中的B4 1 6进行了质谱、1H NMR、13C NMR、1H 13CHMBC、红外光谱等的测定 ,测得分子式为C10 H10 O4 ,推断它为 5,8二羟基 3 甲基 3,4二氢异香豆素。在 2 0 μmol L时 ,它的DPPH自由基捕捉活性为维生素C的 1 67倍 ,维生素E的 2 1倍。 相似文献
2.
枯草芽孢杆菌(Bacillus subtilis)BM9602产生的中性内切β甘露聚糖酶(endoβ1,4Dmannan mannanohydrolase,EC,3.2.1.78)经硫酸铵分级沉淀、DEAE纤维素(DE22)离子交换柱层析,得到电泳纯的样品,提纯了455倍,收率为59%。用SDSPAGE测得该酶的分子量为35kD。用PAGEIEF测得其等电点pI为45。酶反应的最适pH为5.8,最适温度为50℃。该酶在pH60~80,50℃以下稳定。金属离子Hg2+和Ag+对酶活性强烈抑制。酶对槐豆胶、羟丙基瓜胶、田菁胶和魔芋粉的Km值分别为38、149、113和24mg/mL,Vmax值分别为245、865、384和198μmol.min-1mg-1。酶水解甘露聚糖为甘露寡糖(不含单糖)。 相似文献
3.
一株硅酸盐细菌的鉴定及其系统发育学分析 总被引:23,自引:1,他引:23
从南京地区黄棕壤中分离的一株好氧、革兰氏阴性、产芽孢的硅酸盐细菌NBT菌株,能产生丰厚的荚膜,具有鞭毛,能水解淀粉、产生吲哚、液化明胶,全细胞脂肪酸为硬脂酸C16∶0、软脂酸C18∶1(Δ9)和anteisoC15,DNA的G+C mol%为537%。16S rRNA基因测序和系统发育学分析的结果表明,该菌株与胶质芽孢杆菌B7519(Bacillus mucilaginosus)、土壤芽孢杆菌B7517(B. edaphicus)亲缘关系最近。该菌株与B. edaphicus B7517的总DNA杂交率为69%,在形态、生理生化特征上有差异,故可把NBT菌株定为Bacillus edaphicus的一个亚种。 相似文献
4.
从云南西北部土样中分离到一株卡瑞苯西思伯克霍尔德氏菌(Burkholderia caribensis),它在氯乙酸培养基中能产生较高活性的卤乙酸脱卤酶。经硫酸铵盐析、DEAE SephadexA50柱层析、羟基磷灰石柱层析、Sephadex G200 凝胶过滤后,获得电泳纯酶。用SDSPAGE测定酶分子量为46kD。水解氯乙酸的Km值为3.7×10-3mol/L。酶反应的最适温度为40℃,最适pH值为9.5。金属离子及CN-、EDTA对该酶有不同程度的影响,Hg2+和CN-则对该酶有强烈的抑制作用。 相似文献
5.
用离子交换层析(CMsepharose FF)和凝胶层析(SuperdexTM75)方法,从新鲜食用菌毛头鬼伞(Coprinus comatus)子实体中分离纯化出一碱性蛋白y3,经SDSPAGE初步确定其分子量约为14.4kD。活性检测结果显示:当其浓度为12.5μg/mL时,对烟草花叶病毒(TMV)在心叶烟枯斑寄主上的侵染抑制率达83.0%;y3对兔血凝集活性滴度为2.5,对人血凝集活性滴度为26,其浓度分别为1.562μg/mL和0.781μg/mL;利用胃癌细胞株MGC803检测y3体外抗肿瘤活性,其IC50为12μg/mL。y3 N端序列为NRDVAACARFIDDFCDTLTP,为一新的蛋白序列。在SWISSPORT上登录号为P83477。 相似文献
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点状产气单胞菌脯氨酰内肽酶基因的克隆与表达 总被引:2,自引:0,他引:2
用活性筛选法从产气单胞菌点状亚种ST7833
(Aeromonas puctata subsp.puctata ST7833)的基因组中克隆了脯氨酰内肽酶
(Prolyl Endopeptidase,简称apPEP)的基因,测定了含有PEP基因的33kb DNA片段的序列,第202092bp编码了690个氨基酸组成的脯氨酰内肽酶,经检索是一种新的PEP基因。并构建了一株组成性高效表达PEP的基因工程菌BL21/pGEMPEP。BL21/pGEMPEP在
YH培养基中apPEP的表达量占菌体总蛋白的30%左右,活力是野生菌的112倍,表达产物主要为可溶性的胞内蛋白,约5%分泌到胞外。非还原SDSPAGE显示为单体,分子量为76kD,与基因序列预测的分子量一致。试管培养后纯化得到了纯度大于90%的重组脯氨酰内肽酶,比活力为67U/mg。 相似文献
8.
极耐热性阿拉伯糖苷酶基因的表达、纯化及酶学性质研究 总被引:2,自引:0,他引:2
采用PCR从海栖热袍菌(Thermotoga maritima)克隆出编码极耐热稳定性阿拉伯糖苷酶基因,以pET20b为表达质粒,与其C末端6个组氨酸标签序列融合,在大肠杆菌中得到高效表达。基因表达产物通过热处理和亲和层析柱纯化后,酶纯度达电泳均一。纯化重组酶稳定性检测表明,阿拉伯糖苷酶活性最适作用温度和最适作用pH分别为90~95℃和pH 5.0~5.5,在pH 4.2~8.2之间酶活力稳定,95℃的半衰期为4h;SDSPAGE测得酶的分子量为56.57 kD,与理论推算值相吻合。在所测定的底物中,阿拉伯糖苷酶仅对对硝基苯阿拉伯呋喃糖苷(pNPAF)有专一性水解作用,其动力学参数Km值为018mmol/L, Vmax为139μmol/min·mg。 相似文献
9.
甜菜银叶病菌的PCR检测 总被引:5,自引:0,他引:5
本研究用16S23S rDNA间的ITS 序列通用引物L1(5′AGTCGTAACAAGGTAGCCGT3′)和L2 (5′ GTGCCAAGGCATCCACC3)扩增甜菜银叶病菌(Curtobacterium flaccumfaciens pv. betae,Cfb)和其它相近细菌的基因组DNA;并对其PCR产物进行回收、克隆和测序,将所获序列和其它已报道的细菌内源转录间隔区(Internally Transcribed Spacer,ITS)序列进行多重比较后设计出Cfb的特异性引物B1(5′GGCCTCGTGTTGTCCCTTATC3′)和B2 (5′GTCACCAATCAACAACCCGAG3′)。此引物可以从Cfb中扩增出387bp 的特异性片段,而其余参试的21个细菌PCR反应结果均为阴性。该方法可以应用于病害防治工作中的Cfb快速、可靠的检测。 相似文献
10.
阿维链霉菌bkdF的基因中断对阿维菌素合成的影响 总被引:1,自引:0,他引:1
以阿维菌素 B组分菌株Streptomyces avermitilis Bjbm0006为出发菌株,用PCR方法构建支链α酮酸脱氢酶基因bkdF(Branchedchain αketo acid dehydrogenase gene)的重组质粒pHJ5816 (pHZ1358/bkdF&Ermr)对其进行基因中断,得到重组菌株Bjbm5816。经HPLC检测和核磁共振分析发现,Bjbm5816发酵产物产生的单一组分新化合物为OligomycinA。 相似文献
11.
Determination of antioxidant components in rice bran oil extracted by microwave-assisted method 总被引:2,自引:0,他引:2
Rice bran oil was extracted by microwave-assisted extraction with isopropanol and hexane using a solvent-to-rice bran ratio of 3:1 (w/w). The experiments were done in triplicate at 40, 60, 80, 100, and 120 degrees C with a total extraction time of 15 min/sample. The oil components were separated by normal-phase HPLC and quantified with a fluorescence detector. The radical scavenging capability of the oil was tested with DPPH and was expressed as mumol Trolox Equivalent Antioxidant Activity. The increase in total vitamin E with temperature from 40 to 120 degrees C was 59.63% for isopropanol and 342.01% for hexane. Isopropanol was the best solvent for the extraction of gamma-tocopherol and gamma-tocotrienol as compared with hexane for both microwave-assisted and conventional solvent extraction. Isopropanol was better for oil yield extraction at high temperatures. Samples extracted with isopropanol at 120 degrees C had higher antioxidant activity. No differences in oil yield, total vitamin E, and antioxidant activity of oil was noticed between the two methods (microwave-assisted and solvent extractions), at 40 degrees C. No degradation of alpha-tocopherol was noticed during the process. 相似文献
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13.
采用石油醚、乙酸乙酯、乙醇浸提朱红栓菌 Trametes cinnabarina 子实体干粉,得到不同极性提取物;采用清除DPPH 自由基、羟自由基、超氧阴离子自由基能力,测定提取物的体外抗氧化活性;MTT法检测提取物对人肝癌细胞株HepG2细胞增殖的抑制作用。结果表明,朱红栓菌石油醚、乙酸乙酯、乙醇提取物均具有一定的抗氧化、抗肿瘤活性;各提取物在浓度为4-5mg/mL时,对DPPH自由基、羟自由基和超氧阴离子自由基清除能力大小依次为乙酸乙酯提取物>乙醇提取物>石油醚提取物;乙酸乙酯提取物对3种自由基的最高清除率分别为60.23%、74.49%、63.84%。各提取物对人肝癌细胞株HepG2细胞增殖抑制作用大小依次为乙酸乙酯提取物>乙醇提取物>石油醚提取物;乙酸乙酯提取物的抑制率最高达55.93%。采用硅胶和凝胶等柱色谱方法结合核磁、波谱和质谱等技术对乙酸乙酯提取物的化学组分进行分析,共分离纯化出11种化合物,分别鉴定为:麦角甾醇(1),邻苯二甲酸二丁酯(2),对羟基苯甲酸甲酯(3),麦角甾-7,22,二烯-3-酮(4),1-[(12E,16E)-12,16-二十碳二烯酰基]-2-[(E,E)-7,11-十八碳二烯酰基]-3-硬脂酰基甘油(5),cinnabarin(6),过氧麦角甾醇(7),尿嘧啶(8),甘露醇(9),腺嘌呤核苷(10),豆甾-7,22-二烯-3β,5α,6β-三醇(11)。除化合物6外均为首次从朱红栓菌子实体中分离得到。研究结果为开发利用朱红栓菌子实体提供了科学依据。 相似文献
14.
K Li R Xing S Liu Y Qin B Li X Wang P Li 《International journal of biological macromolecules》2012,51(5):826-830
The separation of chitooligomers (COS) with well-defined degree of polymerization (DP) is of interest to further study their bioactivity. However, there has been no report on separation of chitooligomers with DP>6 and the activity of these oligomers is unknown. This paper focuses on separating COS with DP>6 and five fractions were separated from the prepared fully deacetylated chitooligomers mixture by CM Sepharose Fast Flow column and analyzed by HPLC, which mainly contained glucosamine oligomers with DP6-7 (41.31%, 50.22%), DP7-8 (22.47%, 70.13%), DP9-10 (53.06%, 27.99%), DP10-12 (18.45%, 49.36%, 22.31%), and DP>12, respectively. The superoxide radical scavenging activity of each fraction was investigated. The oligomers with DP ranging from 10 to 12 exhibited higher scavenging activity than other fractions and in combination with the DP distribution of fractions, it was further concluded that the chitooligomers with DP11 was likely to be optimal for scavenging superoxide radical activity. 相似文献
15.
TY Chiou TL Neoh T Kobayashi S Adachi 《Bioscience, biotechnology, and biochemistry》2012,76(8):1535-1539
Defatted rice bran extracts were obtained by subcritical treatment using aqueous acetone as extractant. Treatment with 40% (v/v) acetone at 230 °C for 5 min yielded an extract with the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (0.274 mmol of ascorbic acid/g of bran), total carbohydrate (0.188 g/g of bran), protein (0.512 g/g of bran), and total phenolic contents (88.2 mg of gallic acid/g of bran). The effect of treatment temperature (70-230 °C) was investigated using 40% (v/v) acetone, and the extract under 230 °C treatment showed the highest levels of all the determinations described above. The extracts obtained with various concentrations of aqueous acetone were subjected to UV absorption spectra and HPLC analysis, and the results showed changes in composition and polarity. Antioxidative activity evaluated against oxidation of bulk linoleic acid of the extract obtained with 80% (v/v) acetone was higher than that not only of the extract from subcritical water treatment but also of that obtained 40% (v/v) acetone treatment. 相似文献
16.
A large number of polysaccharides are present in boiling-water extraction of Dioscorea nipponica Makino. A DEAE-Sepharose CL-6B column chromatography was used to isolate the major polysaccharides from D. nipponica Makino. The largest amount of fraction of polysaccharide was subjected to further purification by gel-filtration on Sephadex G-100. The purified fraction was a neutral polysaccharide and a single peak in HPLC with Sugar KS-804 column, with a molecular weight of 38,000, and comprised mainly of glucose and fructose (45:1). Analysis by Periodate oxidation–Smith degradation indicated that there were 5.9%(1→)-glycosidic linkages, 4.94% (1 → 2)-glycosidic linkages, 61.16% (1 → 4)-glycosidic linkages, and 28% (1 → 3)-glycosidic linkages. On the basis of superoxide radical assay, hydroxyl radical assay, and self-oxidation of 1,2,3-phentriol assay, its antioxidant activity was investigated. This purified fraction of polysaccharide exhibited equivalent inhibiting power for self-oxidation of 1,2,3-phentriol to Vc, a little higher scavenging activity of superoxide radical and hydroxyl radical than Vc, and should be explored as a novel potential antioxidant. 相似文献
17.
Wiboonsirikul J Kimura Y Kanaya Y Tsuno T Adachi S 《Bioscience, biotechnology, and biochemistry》2008,72(2):384-392
A by-product of rice bran oil and protein production was treated with water and compressed hot water at 20 degrees C to 260 degrees C for 5 min, and at 200 degrees C and 260 degrees C for 5 to 120 min. Each extract was evaluated for its yield, radical scavenging activity, carbohydrate, protein, total phenolic and furfural contents, molecular-mass distribution and antioxidative activity. The maximum yield was obtained at 200 degrees C. The radical scavenging activity and the protein, total phenolic and furfural contents of the extract increased with increasing temperature. However, the carbohydrate content abruptly decreased when treated at above 200 degrees C. The extract treated at 260 degrees C for 5 min exhibited suppressive activity toward the autoxidation of linoleic acid. Each extract obtained at temperatures lower than or equal to 200 degrees C exhibited emulsifying ability. 相似文献
18.
荸荠皮提取物对DPPH自由基清除活性 总被引:13,自引:0,他引:13
采用70%丙酮溶液对荸荠皮中抗氧化物质进行提取,得到红棕色浸膏。通过定性及定量方法分析了荸荠皮提取物中可能存在的具有抗氧化活性的物质;采用DPPH自由基法测定了荸荠皮提取物对DPPH自由基的清除能力。结果显示,荸荠皮提取物中含有多酚类和黄酮类等化合物,其多酚含量为3.31%(w/w,以干物质计)。DPPH自由基法显示,荸荠皮提取物具有一定的清除DPPH自由基能力,其清除能力与提取物浓度之间显示出良好的剂量-效应关系。该提取物(IC50值为130.37 ppm)对DPPH自由基清除能力略低于BHT(IC50值为94.16 ppm)。 相似文献
19.
Safitri R Tarigan P Freisleben HJ Rumampuk RJ Murakami A 《BioFactors (Oxford, England)》2003,19(1-2):71-77
Two antioxidant compounds were isolated from C. sappan L by multiple steps of column chromatography and thin layer chromatography in succession with superoxide scavenging assay as activity monitor. Structures of the two compounds were determined by spectroscopic methods as 1',4'-dihydro-spiro[benzofuran-3(2H),3'-[3H-2]benzopyran]-1',6',6',7'-tetrol (compound 1) and 3-[[4,5-dihydroxy-2(hydroxymethyl) phenyl]-methyl]-2,3-dihydro-3,6-benzofurandiol (compound 2). Characterization of antioxidant properties of these two compounds was done by determining the inhibitory effect on xanthine oxidase activity as well as scavenging effect on superoxide anion and hydroxyl radicals. Our results indicated that compounds 1 and 2 inhibited xanthine oxidase activity and scavenged superoxide anion and hydroxyl radicals. Compounds 1 and 2 possessed similar radical scavenging activities as ascorbic acid, and they were more effective than other well-known antioxidants such as alpha-tocopherol, beta-carotene, and BHT. As inhibitors of free radical formation, compounds 1 and 2 were more effective than all the other antioxidants tested. In conclusion, compounds 1 and 2 can be regarded as primary antioxidants with radical-scavenging and chain-breaking activities as well as secondary antioxidants with inhibitory effect on radical generation. 相似文献