共查询到20条相似文献,搜索用时 0 毫秒
1.
Ethanol-precipitated polysaccharides of the liquid endospermof coconut, Cocos nucifera L., were composed predominantly ofgalactose and arabinose with minor amounts of mannose and glucose.Gel filtration chromatography on Bio-Gel A-0.5 m revealed asingle major peak (Peak A) at the void volume and a minor peak(Peak B) partially included in the column volume. Peak A containedsome uronosyl residues, but was not susceptible to cleavageby endopolygalacturonase, indicating that it does not containsignificant amounts of polygalacturonic acid. Neutral glycosylresidue composition analysis of Peak A showed that it consistedof 72% galactose and 24% arabinose with minor amounts of glucoseand rhamnose. Coconut milk, Cocos nuciferaL, polysaccharides, glycosyl composition 相似文献
2.
Purification and Partial Characterization of a Lectin from Phaseolus vulgaris 总被引:1,自引:1,他引:0
下载免费PDF全文
![点击此处可从《Plant physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
A method is presented for the isolation of a lectin from a Brazilian cultivar of the common bean (Phaseolus vulgaris L.), through extraction in acidic (pH 4.2) medium, fractionation with ammonium sulfate, and chromatography on DEAE-cellulose. The lectin was shown to be homogeneous by gel electrophoresis and isoelectric focusing. 相似文献
3.
采用DEAE阴离子交换层析和Sephadex G100凝胶层析对液体悬浮培养发状念珠藻胞外多糖进行纯化, 得到两个组分NFPS1和NFPS2。对组分NFPS2进行理化性质分析, 并与野生发状念珠藻多糖NFPS0的性质进行对比。结果表明二者具有相似的单糖组成, 均为葡萄糖、木糖、半乳糖、甘露糖; 表观分子量分别为2.79×105、2.26×105; 均不含核酸、蛋白质等物质, 是非硫酸化多糖; 有较高的热稳定性, 其降解温度在245oC左右。但在微观结构上, 两者存在一定差别。 相似文献
4.
发状念珠藻胞外多糖的纯化与性质分析 总被引:1,自引:0,他引:1
采用DEAE阴离子交换层析和Sephadex G100凝胶层析对液体悬浮培养发状念珠藻胞外多糖进行纯化, 得到两个组分NFPS1和NFPS2。对组分NFPS2进行理化性质分析, 并与野生发状念珠藻多糖NFPS0的性质进行对比。结果表明二者具有相似的单糖组成, 均为葡萄糖、木糖、半乳糖、甘露糖; 表观分子量分别为2.79×105、2.26×105; 均不含核酸、蛋白质等物质, 是非硫酸化多糖; 有较高的热稳定性, 其降解温度在245oC左右。但在微观结构上, 两者存在一定差别。 相似文献
5.
The microbial production of dextranase using cheap carbon sources is beneficial to solve the economic loss caused by the accumulation of dextran in syrup. A food-grade microbial cell factory was constructed by introducing the dextranase encoding gene DEX from Chaetomium gracile to the chromosome of Bacillus subtilis, and the antibiotic resistance marker gene was subsequently deleted via the Cre/loxP strategy. The dual-promoter system with a sequentially arranged constitutive P43 promoter resulted in an 85 % increase in DEX expression. Under the optimal fermentation conditions of 10 g/L maltose, 15 g/L casein, 1 g/L Na2HPO4, 1 g/L FeSO4 and 8 g/L NaCl, DEX activity was increased from 2.625 to 64.34 U/mL. Recombinant DEX was purified 5.98-fold with a recovery ratio of 26.67 % and specific activity of 3935.02 U/mg. Enzyme activity was optimal at 55 °C and pH 5.0 and remained 80.34 % and 71.36 % of the initial activity at 55 °C and pH 4.0 after 60 min, respectively. The enzyme possessed high activity in the presence of Co2+, while Ag+ showed the strongest inhibition ability. The optimal substrate was 20 g/L dextran T-2000. The findings could facilitate the low-cost, large-scale production of food-grade DEX for use in the sugar industry. 相似文献
6.
从假菠萝麻叶汁中用乙醇分部沉淀法得到一种蛋白酶,对酪蛋白有强烈的水解活性,经Sephadex G-100柱层析和SP-Sephadex C-50离子交换柱层析等步骤纯化,可得到六角形结晶。结晶酶液经PAGE圆盘电泳,每条胶柱上只显示一条蛋白带,其活性在pH4。5-10、55℃范围内较稳定,最适pH8.5最适温度50℃,Km(酪蛋白)值为0.185%(W/V)。用SDS-PAGE和Sephadex 相似文献
7.
A nonganglioside factor(s) present in Sigma types II and III mixed bovine brain ganglioside preparations synergises with suboptimal amounts of serum to induce proliferation specifically in nondividing B 103 neuroblastoma cultures. The active substance is nondialysable and soluble in water as well as in chloroform-methanol mixtures of 1:1-4:1 (vol/vol). It is completely insoluble in ether and acetone at room temperature. Biological activity survives heating to 70 degrees C in the presence of 0.1 M HCl for 1 h as well as boiling at neutral pH. Loss of activity occurs on heating to 70 degrees C for 1 h with 1 M HCl or 1 M NaOH. The activity is insensitive to digestion with neuraminidase, trypsin, pronase, and phospholipases A2 and C. The factor cochromatographs with gangliosides on Dowex AG 50W and Sephadex G100 and is partially recovered with GM1 on DEAE-Sepharose, but may be isolated in a ganglioside-free fraction by sequential chromatography on Sephadex LH20 and silicic acid columns. The substance(s) has the properties of a water-soluble proteolipid protein, the amino acid composition being reported. It is not immunologically cross-reactive with antibodies to GM1 ganglioside or the major proteolipid protein of myelin. 相似文献
8.
Partial Purification and Characterization of Hemolysin from a Psychrotrophic Kanagawa-Positive Marine Vibrio 总被引:2,自引:1,他引:1
下载免费PDF全文
![点击此处可从《Applied microbiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Psychrotrophic Kanagawa-positive marine vibrios were isolated from soft-shelled clams (Mya arenaria) collected in Yaquina Bay, Oreg. The 235 vibrio isolates obtained were screened for Gram reaction and morphology, Kanagawa reaction on Wagastsuma agar, and response to selected biochemical tests. The vibrio selected for further study was grown in broth, and the hemolysin was precipitated from a cleared supernatant with solid ammonium sulfate. The hemolytic substance was partially purified by DEAE-cellulose and Sephadex G-100 column chromatography. The hemolysin contained protein essential for activity, was thermolabile, and was more active against rabbit erythrocytes at 37°C than at lower temperatures. The molecular weight was estimated at 55,000 by using a Sephadex G-100 column. Hemolytic activity was partially inactivated by gangliosides and lowered against horse erythrocytes. The hemolysin did not react with antibody prepared against vibriolysin from Vibrio parahaemolyticus WP-1 by the Ouchterlony method. The hemolysin was high in aspartic and glutamic acids and low in arginine and histidine. Electrophoresis on a sodium dodecyl sulfate-polyacrylamide gel gave three major bands. The hemolysin from a psychrotrophic vibrio and the hemolytic exotoxin of V. parahaemolyticus had some similar and dissimilar characteristics. The possibility that a Vibrio sp. other than V. parahaemolyticus might serve as the reservoir for the Kanagawa phenotype is discussed. 相似文献
9.
A thermostable amylase, possibly a beta-amylase from Thermoactinomyces sp. no. 2 isolated from soil, is reported. The enzyme was purified 36-fold by acetone precipitation, ion-exchange chromatography, and Sephadex G-200 gel filtration, and the molecular weight was estimated at 31,600. The enzyme was characterized by demonstration of optimum activity at 60 degrees C and pH 7 and by retention of 70% activity at 70 degrees C (30 min). It was stimulated by Mn and Fe but strongly inhibited by Hg. Maltose was the only detectable product of hydrolysis of starches and was quantitatively highest in plantain starch hydrolysate. 相似文献
10.
Carlos A. Bonilla Reed M. Stringham Jr. 《Preparative biochemistry & biotechnology》2013,43(4):305-328
This study reports the isolation and partial purification of a polypeptide from human saliva which causes a significant serum calcium lowering when administered to mice. Purification was achieved by preparative electrophoresis, dialysis, two gel filtration steps on Sephadex G-150, and ion exchange chromatography on DEAE-cellulose. Homogeneity was determined by poly-acrylamide electrophoresis. Blood sampling was carried out by puncture of the orbital venous plexus and serum analyzed for calcium. The most active preparations lower serum calcium from 10–27% of initial value, producing tetany and convulsions in some cases. The molecular weight of this polypeptide was estimated to be 4, 260 by the use of a calibrated Sephadex G-75 column. This is a much smaller molecular weight than that expected from its initial exclusion from Sephadex G-150, and suggests that this hypocalcemic factor is associated with larger molecules through most of the purification procedure up to and including DEAE-cellulose chromatography. A second gel filtration on Sephadex G-150 separates two minor salivary protein contaminants (IgA and IgG immunoglobulin) in the excluded fraction from the smaller, hypocalcemically active polypeptide. No hypocalcemia activity could be detected or isolated in a preliminary investigation on the saliva of a dysgammaglobuli-nemic (IgA deficient) patient. The hypocalcemia induced does not differ significantly from that observed after administration of calcitonin to mice in that: 2) minimum values are reached in 1.5–2 hours and return to normal in 5–6 hours, b) magnitude of hypocalcemia response is dose dependent. The salivary hypocalcemia factor isolated in this study has the properties of a protein, in that its activity is destroyed by the proteolytic enzyme trypsin, it yields amino acids upon acid hydrolysis and it behaves on electrophoresis, gel filtration and ion exchange chromatography as a typical protein. 相似文献
11.
KazuhirÖ Ichikawa Kozaburo Okazaki Kazuhisa Kimoto Yasuto Watanabe 《Bioscience, biotechnology, and biochemistry》2013,77(5):1297-1299
The structure of αsl -casein at an emulsified oil/water interface was studied by immunochemical method using BALB/c mouse anti-αsl -casein antibody. By incubating the antiserum with an sl-casein-stabilized emulsion, all of the specific antibodies in the serum were absorbed by the emulsion, suggesting that αsl-casein was adsorbed to the oil surface, all of its antigenic determinants being exposed to the aqueous phase. Since the antigenic determinants of αsl-casein for BALB/c mice are known to be present in the regions of residues 1 ~ 8, 33 ~ 54,105 ~ 119,133 ~ 151, and 174 ~ 199, these portions must be, at least partly, exposed to the aqueous phase and accessible to the antibodies. 相似文献
12.
ABSTRACT Ichthyophthirius multifiliis , a parasitic ciliate of freshwater fishes, was found to have surface antigens (Ag) which elieited immobilizing antibodies (Ab) when injected into rabbits. An effort was made to purify and characterize these Ag (referred to as immobilization Ag) because of their potential role in protective immunity in fishes. Mice immunized with theront cilia were used for production of immobilizing monoclonal antibodies (MAb). Hybridomas were screened by indirect immunofluorescent light microscopy and immobilization of live parasites. Six hybridomas producing immobilizing MAb were cloned. Immobilizing MAb were used to affinity purify Ag solubilized with Triton X-114 and Na deoxycholate. Two membrane protein Ag of approximately 48 and 60 kDa were identified. Immobilizing MAb failed to react with these Ag on Western blots and, conversely, MAb that reacted with the Ag on Western blots did not immobilize live organisms. These results suggest that immobilization required native conformational epitopes which were altered by Western blotting procedures. Individual MAb reactive on Western blots recognized both the 48- and 60-kDa proteins indicating the presence of common epitopes. Affinity purified Ag elicited immobilizing antisera when injected into rabbits, mice, and channel catfish. 相似文献
13.
Aminopeptidase II, one of the two major aminopeptidases in the giant alga Chara australis, was partially purified. Its molecular weight was estimated to be about 80,000 by gel permeation chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that it is composed of a single polypeptide with a molecular weight of about 85,000. Aminopeptidase II hydrolyzed alanine-2-naphthylamide more efficiently than the naphthylamides of lysine and proline, and only weakly hydrolyzed the naphthylamides of arginine, phenylalanine, valine, and leucine. The optimal pH for the hydrolysis of alanine-2-naphthylamide was near 7.0. The activity of aminopeptidase II was inhibited by the SH-reagents p-chloromercuribenzoic acid and N-ethylmaleimide and by the metal chelator 1,10-phenanthroline. 相似文献
14.
Sankar Das Tirtha J. Bhaduri Anindita Bose Bharati Ghosh 《Journal of plant biochemistry and biotechnology.》1996,5(2):123-126
Arginine decarboxylase (EC 4.1.1.19) has been purified and characterized from Brassica campestris cv B-9. The enzyme was purified 1120 fold and the recovery was 9%. The mol wt of the enzyme determined by gel filtration was 240 kD with identical subunits of 60 kD. The pH and temperature optima for the enzyme were 8.0 and 30°C respectively. The Km was 0.31mM. Polyamines inhibited the enzyme activity significantly. Immunodiffusion with ADC-specific antibodies showed cross reactivity against purified ADC from Brassica. 相似文献
15.
A surface-bound aminopeptidase of Lactobacillus lactis cells was solubilized with lysozyme, and the extract was subjected to streptomycin sulfate precipitation, ammonium sulfate fractionation, chromatography on Sephadex G-100 and diethylaminoethyl-Sephadex A-50, and preparative polyacrylamide gel electrophoresis. The purified enzyme was homogeneous in disc electrophoretic analysis and consisted of a single polypeptide chain with a molecular weight of 78,000 to 81,000. The optimal pH and optimal temperature for enzyme activity were 6.2 to 7.2 and 47.5 degrees C, respectively, for l-lysine-4-nitroanilide as the substrate. The enzyme was activated by Co and Zn ions and inhibited by Cu, Hg, and Fe ions and by the metal-complexing reagents ethylenediaminetetraacetic acid, 1,10-phenanthroline, and alpha,alpha'-dipyridyl. Higher concentrations of substrate and hydrolysis products also inhibited the activity of the enzyme. The aminopeptidase had broad substrate specificity and hydrolyzed many amino acid arylamides and many peptides with unsubstituted NH(2)-terminal amino acids. 相似文献
16.
中国对虾(Penaeus chinensis)酚氧化酶的分离纯化及其部分生物化学性质 总被引:25,自引:0,他引:25
以中国对虾 (Penaeuschinensis)的血淋巴为材料 ,利用凝胶过滤和离子交换等方法 ,对酚氧化酶 (phe noloxidase ,E .C .1 .1 0 .3.1 )进行了分离纯化和生物化学性质研究。实验发现 ,酚氧化酶原 (prophenoloxidase)的分子量为 87.5kD左右 ,在实验操作过程中极易发生降解或自身降解 ,变成有活性的大小约 77kD的酚氧化酶 ,1 0g/LSDS可使该酶原全部被激活而成为有活性的酚氧化酶。以L 二羟苯丙氨酸 (L DOPA)为特异性底物对酚氧化酶活性进行研究发现 ,其最适pH值为 6 .0左右 ,最适温度为 4 0℃ ,Km 值约为 1 .99mmol/L。利用多种氧化酶抑制剂对酚氧化酶纯化样品的酶活性进行研究 ,发现抗坏血酸 (ascorbicacid)、半胱氨酸 (cysteine)和二硫苏糖醇(dithiothreitol)对酚氧化酶活性具有很强的抑制作用 ,硫脲 (thiourea)对酚氧化酶活性也具有较强的抑制作用 ,而酚氧化酶对苯甲酸 (benzoicacid)、柠檬酸 (citricacid)和亚硫酸钠 (sodiumsulfite)不敏感 ,而且该酶对酪氨酸等单酚还具有高特异性的酚氧化酶活性 ,表明它可能是一种酪氨酸酶型的酚氧化酶 ;此外 ,该酶对EDTA和金属离子非常敏感 ,其活性能被Cu2 强烈抑制 ,被Mg2 强烈激活 ,表明该酶很可能是一种金属酶 (metalloenzyme) 相似文献
17.
Ming Xing Huang Yun Ye Ya Xiong Chen Ya Li Han 《International journal of peptide research and therapeutics》2012,18(2):153-161
Two proteins with fibrinolytic activity were partially purified from yellow mealworm (Tenebrio molitor) by ammonium sulfate precipitation between 30 and 70% saturation, gel filtration on Sephacryl-S200-HR, ion exchange chromatography
on DEAE-Sepharose-FF and metal chelate on Cu–HiTrap–IMAC–FF, but the enzymes had not been completely separated from each other.
The two partially purified fibrinolytic enzymes were designated as TMFE-I and TMFE-II (Tenebrio molitor fibrinolytic enzyme) with molecular weights of 27.5 and 24.9 kDa by SDS-PAGE individually. The partially purified solution
of TMFE-I and TMFE-II was considerably stable in the range of pH 5–10 and characterized by pH optimum of the enzymatic activity
at 8.0. Thermal stability of TMFE was excellent at 45°C and below. The K
M value was 0.26 mM for amidolysis of Bz–Arg–pNA. According to inhibitor analysis by fibrin plate method, phenylmethylsulfonyl
fluoride and tosyl-lysine chloromethyl ketone inactivated TMFE almost completely, but trans-(epoxysuccinyl)-l-leucylamino-4-guanidinobutane (E-64) and EDTA had little effect on their fibrinolytic activity. According to metal ion analysis
by fibrin plate method, the effect of metal ions on activity of TMFE showed a great difference. Na+, K+ and Zn2+ had little effect on the activity of TMFE. Mg2+ and Cu2+ showed inhibition effect on the fibrinolytic activity of TMFE, but Ca2+ increased the fibrinolytic activity of TMFE at final concentration varying from 0 to 30 mM. 相似文献
18.
Purification and Partial Characterization of a Prolyl-Dipeptidyl Aminopeptidase from Lactobacillus helveticus CNRZ 32 总被引:8,自引:6,他引:2
下载免费PDF全文
![点击此处可从《Applied microbiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
X-prolyl-dipeptidyl aminopeptidase, which hydrolyzed Gly-Pro-p-nitroanilide (relative activity [RA] = 100%) and Arg-Pro-p-nitroanilide (RA, 130%), was purified to homogeneity from the cell extract of Lactobacillus helveticus CNRZ 32. The enzyme also hydrolyzed Ala-Pro-Gly (RA, 11%) and Ala-Ala-p-nitroanilide (RA, 2%) but was not active on Ala-Leu-Ala, dipeptides, and endopeptidase and carboxypeptidase substrates. The enzyme was purified 145-fold by streptomycin sulfate precipitation, ammonium sulfate fractionation, and a series of column chromatographies on DEAE-cellulose, arginine-Sepharose 4B, and glycyl-prolyl-AH-Sepharose 4B. The purified enzyme appeared as a single band on native polyacrylamide gel and sodium dodecyl sulfate-polyacrylamide gel electrophoreses and had a molecular weight of 72,000. Optima for activity by the purified enzyme were pH 7.0 and 40°C. The enzyme was incubated at 40°C for 15 min with various metal ions. It was activated by Mg2+ (2.5 mM), Ca2+ (0.1 to 2.5 mM), Na+ (10 to 50 mM), and K+ (10 to 50 mM) and was inhibited by Hg2+ (0.1 to 2.5 mM), Cu2+ (0.1 to 2.5 mM), and Zn2+ (0.1 to 2.5 mM). Enzyme activity was partially inhibited by EDTA (1.0 mM, 20 h at 40°C), 1,10-phenanthroline (1.0 mM, 15 min at 40°C), phenylmethylsulfonyl fluoride (1.0 mM), N-ethylmaleimide (1.0 mM), and iodoacetate (1.0 mM). It was completely inhibited by diisopropyl fluorophosphate (1.0 mM, 2 h at 40°C) and p-chloromercuribenzoate (1.0 mM, 15 min at 40°C). The enzyme was not affected by dithioerythritol (1.0 to 10 mM). 相似文献
19.
An exocellular proteinase synthesized by the geophilic dermatophyte Trichophyton vanbreuseghemii has been purified and characterized. The fungus obtained from soil in Iran was cultivated in modified Czapek–Dox liquid medium
containing 0.1% bacteriological peptone and 1% glucose as the nitrogen and carbon sources. Partial purification of the proteinase
was accomplished by (NH4)2SO4 precipitation, followed by ion exchange chromatography. Analysis of the enzyme by SDS-PAGE revealed a single polypeptide
chain with an apparent molecular mass of 37 kDa. Proteinase activity was optimum at pH 8, but remained high in the range of
pH 7–11. Moreover, the partially purified enzyme presented a keratinolytic activity as evidenced by the keratin azure test.
The inhibition profile and the good activity of the enzyme towards the synthetic substrate N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide suggested that it belonged to the chymotrypsin/subtilisin group of serine proteinases. The keratinolytic properties
of T. vanbreuseghemii suggest that this fungus may be an alternative for the recycling of industrial keratinic wastes. 相似文献
20.
Characterization and Partial Purification of a Novel Neuronotrophic Factor from Bovine Seminal Vesicle 总被引:1,自引:1,他引:0
Extracts from bovine seminal vesicles have been shown to contain high concentrations of nerve growth factor (NGF)-like biological activity and of the NGF protein with properties corresponding to that of NGF from other sources. We now demonstrate that a second neuronotrophic protein, termed seminal vesicle-derived neuronotrophic factor (SVNF), is present in seminal vesicle extracts (SVEs), which could not be distinguished from NGF on the basis of biological activity. SVNF has neuronotrophic activity on NGF target cells like embryonic chicken-sensory and sympathetic neurons, sympathetic neurons, and chromaffin cells from neonatal rats, but it is inactive on embryonic chicken ciliary or neonatal rat nodose ganglion neurons. It also stimulates fiber outgrowth from rat pheochromocytoma (PC 12) cells. In gel filtration chromatography on Biogel A 1.5 m, the activity is eluted with an apparent molecular weight of 40 kilodaltons, and by preparative isoelectric focusing, the isoelectric point was determined to be in the neutral range (6.8-7.8). The biological activity of SVNF, in contrast to that of NGF, is partially retained after preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and can be electrophoretically eluted with an apparent molecular weight of 16-20 kilodaltons. Electrophoretically purified SVNF is not inhibited by antisera to mouse NGF, but its activity is increased greater than 10-fold in the presence of very low concentrations of NGF. For partially purified SVNF, a specific activity of 2.9-5.8 X 10(5) biological units/mg of protein was determined in the presence of subthreshold NGF concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献