scholarly journals Identification and characterization of diverse xylanases from thermophilic and thermotolerant fungi

BioResources ◽  
2006 ◽  
Vol 1 (1) ◽  
pp. 18-33 ◽  
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Catalytic Activities ◽  
Humicola Insolens ◽  
Thermotolerant Fungi ◽  
Chaetomium Thermophilum ◽  
Isoelectric Points ◽  
Birch Wood Xylan ◽  
Functionally Diverse ◽  
Identification And Characterization

Thirteen fungal isolates included in this study expressed multiple xylanase isoforms as observed by xylan zymograms of polyacrylamide gel electrophoresis (PAGE) and isoelectrofocussing (IEF) fractionated proteins. Eighty-three xylanases produced by these thermophilic and thermotolerant strains were detected using the IEF profiling technique. Xylanases identified on the basis of their isoelectric points (pI) were functionally diverse and exhibited differential catalytic activities against various xylan types (birch wood xylan, larch wood xylan, oat spelt xylan, rye arabino xylan and wheat arabino xylan) as well as debranched arabinan. Thermophilic isolates, Chaetomium thermophilum , Humicola insolens , Melanocarpus sp., Malbranchea sp. and Thermoascus aurantiacus , were found to produce alkaline active xylanases that showed a bleach boosting effect on Decker pulp resulting in increased brightness (1.60-2.04 ISO units).

Identification and characterization of cDNA clones encoding two homologous proteins that are part of the asialoglycoprotein receptor

1987 ◽  
Vol 7 (5) ◽  
pp. 1841-1847
Author(s):  
M McPhaul ◽  
P Berg
Keyword(s):  
Rat Liver ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Asialoglycoprotein Receptor ◽  
Cdna Clones ◽  
Differential Splicing ◽  
Homologous Proteins ◽  
Different Types ◽  
Identification And Characterization

The asialoglycoprotein receptor (ASGP-R) from rat liver contains the following three distinct protein species when it is analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis: RHL1 (42 kilodaltons), RHL2 (49 kilodaltons), and RHL3 (54 kilodaltons). In this paper we describe the isolation of cDNA clones encoding RHL1 and RHL2 from a cDNA library constructed from rat liver mRNA. A comparison of the predicted coding sequence for RHL2 with that for RHL1 showed that these sequences are highly homologous. The library also contained numerous cDNA clones for both RHL1 and RHL2 that were derived from unspliced precursor mRNAs. Differential splicing at the 5' end of the RHL1 transcript was inferred from the finding that two different types of RHL1 cDNA were identified, each having a different 5' terminus.

scholarly journals Studies of the human factor VIII/von Willebrand's factor protein. II. Identification and characterization of the von Willebrand protein

Blood ◽  
1975 ◽  
Vol 46 (3) ◽  
pp. 417-430
Author(s):  
HR Gralnick ◽  
BS Coller
Keyword(s):  
Factor Viii ◽  
Human Factor ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Related Protein ◽  
Human Factor Viii ◽  
Von Willebrand’S Factor ◽  
Von Willebrand ◽  
Identification And Characterization

The purified factor VIII-related protein we have previously characterized from normal cryoprecipitate possesses both procoagulant activity and vWf activity. We have attempted to isolate and characterize this protein from three patients with severe vWd. This protein is absent or markedly diminished in amount in these vWd patients, as judged by gel filtration, polyacrylamide-gel electrophoresis, and immunoprecipitation assays. Likewise, the procoagulant and vWf activities are deficient. As vWf activity is one of the major biologic functions of either the normal or hemophilic factor VIII-related protein, the purified protein should be designated the f VIII/vWf protein.

Identification and characterization of a pancreatic intrinsic factor in the dog

1989 ◽  
Vol 256 (3) ◽  
pp. G517-G523 ◽  
Author(s):  
R. M. Batt ◽  
N. U. Horadagoda ◽  
L. McLean ◽  
D. B. Morton ◽  
K. W. Simpson
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Intrinsic Factor ◽  
Gel Filtration ◽  
Polyclonal Antiserum ◽  
Pure Pancreatic Juice ◽  
Intrinsic Factors ◽  
Rabbit Polyclonal Antiserum ◽  
Identification And Characterization

An intrinsic factor has been identified in the canine pancreas, and output and properties of this protein have been compared with those of gastric intrinsic factor in the dog. Mean concentrations of intrinsic factor and peak outputs per minute were approximately 5- to 10-fold higher in pure pancreatic juice after stimulation with secretin and cholecystokinin, respectively, than in pentagastrin-stimulated gastric juice. Purified gastric and pancreatic intrinsic factors had an identical molecular mass of 65 kDa, estimated by gel filtration on Sephacryl S-200, while sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated single bands corresponding to 53 kDa. Immunoblots showed that rabbit polyclonal antiserum to canine gastric intrinsic factor cross-reacted with canine pancreatic intrinsic factor. Gastric and pancreatic intrinsic factor-cyano[57Co]cobalamin complexes exhibited comparable association constants for ileal receptors in canine brush-border vesicles, while there was minimal binding to jejunal vesicles. These findings demonstrate that the canine pancreas is an important source of an intrinsic factor that closely resembles gastric intrinsic factor in the dog.

scholarly journals Isolation and characterization of lung connective-tissue glycoproteins

1982 ◽  
Vol 203 (3) ◽  
pp. 593-601 ◽  
Author(s):  
C Lafuma ◽  
M Moczar ◽  
L Robert
Keyword(s):  
Proteinase Inhibitors ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Lung Parenchyma ◽  
Isolation And Characterization ◽  
Isoelectric Points ◽  
And Function

1. Glycoproteins of hamster, rat and baboon lung parenchyma were investigated by using [14C]glucosamine incorporation in vitro followed by sequential extraction of the macromolecular components and characterization of the glycoproteins in the extracts. 2. Slices of lung parenchyma maintained in vitro incorporated [U-14C]glucosamine linearly with time into non-diffusible macromolecules for up to 5h. All the macromolecule-associated 14C label was present as [14C]glucosamine. 3. These 14C-labelled macromolecules were extracted from previously delipidated and salt-extracted lung by 5M-guanidinium chloride in the presence of dithiothreitol and proteinase inhibitors before (extract A1) and after (extract A2) hydrolysis of the collagen by collagenase. The [14C]glucosamine-labelled glycoproteins in extracts A1 and A2 contained 55 and 5% respectively of the total [14C]glucosamine incorporated in the lung of all three species studied. 4. The [14C]glucosamine-labelled glycoproteins were analysed by gel-filtration chromatography, sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and isoelectric focusing. The major [14C]glucosamine-labelled glycoproteins of baboon lung parenchyma had apparent mol.wts. of about 400 000, 140 000 and 65 000 with isoelectric points respectively of 4.8, 5.4 and 5.4. The hamster lung glycoproteins with isoelectric points of 4.1 and 5.8 were devoid of hydroxyproline and contained galactose, mannose and N-acetylglucosamine. These experiments indicate that several distinct glycoproteins are synthesized in situ by the cells of pulmonary parenchyma and may well play a role in its structure and function.

scholarly journals Identification and characterization of cDNA clones encoding two homologous proteins that are part of the asialoglycoprotein receptor.

1987 ◽  
Vol 7 (5) ◽  
pp. 1841-1847 ◽  
Author(s):  
M McPhaul ◽  
P Berg
Keyword(s):  
Rat Liver ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Asialoglycoprotein Receptor ◽  
Cdna Clones ◽  
Differential Splicing ◽  
Homologous Proteins ◽  
Different Types ◽  
Identification And Characterization

The asialoglycoprotein receptor (ASGP-R) from rat liver contains the following three distinct protein species when it is analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis: RHL1 (42 kilodaltons), RHL2 (49 kilodaltons), and RHL3 (54 kilodaltons). In this paper we describe the isolation of cDNA clones encoding RHL1 and RHL2 from a cDNA library constructed from rat liver mRNA. A comparison of the predicted coding sequence for RHL2 with that for RHL1 showed that these sequences are highly homologous. The library also contained numerous cDNA clones for both RHL1 and RHL2 that were derived from unspliced precursor mRNAs. Differential splicing at the 5' end of the RHL1 transcript was inferred from the finding that two different types of RHL1 cDNA were identified, each having a different 5' terminus.

scholarly journals The isolation and characterization of corticotropin from the pituitary gland of the ostrich Struthio camelus

1977 ◽  
Vol 165 (3) ◽  
pp. 519-523 ◽  
Author(s):  
R J Naudé ◽  
W Oelofsen
Keyword(s):  
Amino Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Amino Acid Residues ◽  
Struthio Camelus ◽  
Isolation And Characterization ◽  
Isoelectric Points ◽  
Pituitary Glands ◽  
Amide Content

1. Avian corticotropin (ACTH) was purified from both fresh and aged pituitary glands of the ostrich Struthio camelus. 2. The isolation of corticotropin in pure form involved acid/acetone extraction, NaCl fractionation, CM-cellulose chromatography and Sephadex G-50 chromatography. 3. The hormone preparations from fresh and aged glands behaved as single substances on polyacrylamide-gel electrophoresis, and both preparations were found to consist of 39 amino acid residues, in identical molar proportions for the different amino acids. 4. The isoelectric points of the two hormone preparations were estimated to be in the range pH 8.3-8.7, indicating possible differences in amide content, and the N-terminal amino acid of both preparations appeared to be serine. 5. The hormone preparations from fresh and aged glands exhibited similar biological potencies (73 and 77 i.u./mg respectively), as measured by steroidogenesis in vitro. 6. Apart from possible differences in amide content, the corticotropin preparations obtained from fresh and aged glands appear to be indistinguishable.

Characterization of bacteriocin 28 produced by Clostridium perfringens

1982 ◽  
Vol 28 (7) ◽  
pp. 860-873 ◽  
Author(s):  
A. W. Li ◽  
J. A. Verpoorte ◽  
R. G. Lewis ◽  
D. E. Mahony
Keyword(s):  
Clostridium Perfringens ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Active Material ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Hydrophobic Properties ◽  
Isoelectric Points

Bacteriocin 28, produced by Clostridium perfringens, was characterized by gel filtration and sodium dodecyl sulfate – polyacrylamide gel electrophoresis as a glycoprotein with a molecular weight of approximately 100 000. Density gradient centrifugation suggested a lower weight of 84 000. The bacteriocin bound firmly to phenyl-Sepharose CL-4B gel, indicating hydrophobic properties, and elution from this gel with ethylene glycol clearly separated bacteriocin from the alpha and theta toxins of C. perfringens, the latter of which was also hydrophobic. Bacteriocin 28 was immunogenic, inducing neutralizing and precipitating antibodies, and possessed three isoelectric points: 7.37, 7.05, and 5.4. Amino acid and carbohydrate analysis of the active material showed a composition of 15 amino acids and several carbohydrates. The molecule demonstrated instability with increasing purification, and several approaches to purification are described.

Production and Characterization of Ovalbumin and Ovotransferrin from Chicken Egg White

2012 ◽  
Vol 8 (2) ◽  
Author(s):  
Jianguo Liu ◽  
Jing Liu ◽  
Xuefang Zhang
Keyword(s):  
Amino Acid ◽  
Binding Capacity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Egg White ◽  
Molecular Weights ◽  
Chicken Egg ◽  
Isoelectric Points ◽  
Iron Binding Capacity

An efficient and easily scaled up method to separate ovalbumin and ovotransferrin simultaneously from chicken egg white using ultrafiltration is proposed. The purities of ovalbumin and ovotransferrin obtained were 94% and 89%, with the yields of 82% and 76%, respectively. The resulting ovalbumin and ovotransferrin products was then characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isoelectric focusing, dynamic light scattering, circular dichroism, and amino acid analysis to confirm their molecular weights, isoelectric points, aggregate sizes, molecular secondary structures, and amino acid compositions. The iron-binding capacity of the purified ovotransferrin was also evaluated.

scholarly journals Electrophoretic Characterization of Venom Proteins of Montivipera xanthina (Gray, 1849) (Ophidia: Viperidae)

2017 ◽  
Author(s):  
Hüseyin Arıkan ◽  
Nurşen Alpagut Keskin ◽  
Kerim Çiçek
Keyword(s):  
Serine Proteases ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sds Page ◽  
Dependent Activity ◽  
Fibrinogenolytic Activity ◽  
Identification And Characterization ◽  
Insight Into ◽  
Venom Proteins

In this study, with the aim of evaluating coagulant activities in the venom of M. xanthina, we analyzed venom proteins, digestion patterns of fibrinogen chains with venom and the effects of protease inhibitors on M. xanthina venom proteases using Tricine-sodium dodecyl sulfate polyacrylamide gel electrophoresis. Venom samples were obtained from four adult specimens of Montivipera xanthina collected in Gümüldür (Izmir, Turkey). SDS-PAGE analysis demonstrated that 17 protein bands in the range of 20–250 kDa were present. The specific digestion patterns of fibrinogen chains revealed that M. xanthina venom possesses fibrinogenolytic enzymes which could be included in coagulation processes during envenomation Fibrinogenolytic activity directed exclusively towards the Aa-chain with a time-dependent activity towards Bb-chains suggests the presence of both metalloproteinases and serine proteases in M. xanthine venom. In the present study, the occurrence and inhibition of fibrinogenolytic activity of M. xanthina venom were clearly observed. For further analysis, the isolation, identification, and characterization of individual venom components will provide insight into their function and biological roles.

scholarly journals Isolation and characterization of three Ca2+-dependent β-galactoside-specific lectins from snake venoms

1984 ◽  
Vol 224 (1) ◽  
pp. 301-307 ◽  
Author(s):  
T K Gartner ◽  
M L Ogilvie
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Reducing Agents ◽  
Agkistrodon Contortrix ◽  
Isolation And Characterization ◽  
Isoelectric Points ◽  
Diamondback Rattlesnake ◽  
Agkistrodon Contortrix Contortrix ◽  
Bothrops Atrox

Three lactose-inhibited lectins from the venoms of the snakes Agkistrodon contortrix contortrix (southern copperhead), Ancistrodon piscivorous leukostoma (western cottonmouth moccasin) and Crotalus atrox (western diamondback rattlesnake) have been isolated and newly characterized. The three lectins are similar to thrombolectin, a lectin isolated from the venom of Bothrops atrox (fer-de-lance) (Gartner, Stocker & Williams, 1980), with regard to sugar specificity, Mr, Ca2+ requirements and sensitivity to reducing agents. Each lectin is a dimer (Mr 28 000) consisting of monomers (Mr 14 000) indistinguishable on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Haemagglutination activity is dependent on the presence of Ca2+ and is inhibited by reducing agents. The lectins are not identical and can be distinguished on the basis of relative affinities for inhibiting sugars, isoelectric points and immunoprecipitation assays using anti-(cottonmouth lectin) serum.

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