Isolation and Characterization of Viridin, a New 65 kDa Antifungal Protein from the Mould Trichoderma viride

1999 ◽  
Vol 380 (10) ◽  
pp. 1243-1245 ◽  
Author(s):  
Jian-Jiang Hao ◽  
Chuan-dong Geng ◽  
Wei-jun Xie ◽  
Zhen-zhen Gong ◽  
Wang-Yi Liu ◽  
...  
Keyword(s):  
High Performance ◽  
Culture Medium ◽  
Basic Protein ◽  
Trichoderma Viride ◽  
Exchange Chromatography ◽  
Antifungal Protein ◽  
Cation Exchange Chromatography ◽  
Isolation And Characterization ◽  
Sds Page

AbstractA new extracellular antifungal protein with a yield of 10 mg per liter was isolated from the culture medium of the mouldTrichoderma viride. The protein, which we named viridin, was purified by carboxymethyl-cellulose cation-exchange chromatography and Superose 12 HR 10/30 high-performance liquid chromatography. Viridin, a basic protein of approximately 65 kDa as determined by SDS-PAGE, inhibits the growth of the cotton pathogenVerticillum dahliae, the IC50being 6 ΜM.

scholarly journals ISOLASI DAN KARAKTERISASI ENZIM b-1,3-ENDOGLUKANASE DARI TANAMAN KUBIS (Brassica oleracea cv. Capitata L.)

2010 ◽  
Vol 15 (2) ◽  
pp. 99-105
Author(s):  
Y. Sri Manuhara
Keyword(s):  
Ion Exchange ◽  
Brassica Oleracea ◽  
Hydrophobic Interaction Chromatography ◽  
Inhibition Zone ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Isolation And Characterization ◽  
Sds Page ◽  
Optimum Ph

Isolation and characterization of β-1,3-endoglucanase from cabbage (Brassica oleracea var. capitata L.) have been done. It showed 40° C of optimum temperature, and optimum pH is 7. After the purification with hydrophobic interaction chromatography and ion exchange chromatography, it’s activity was increased. Based on SDS-PAGE analysis, β-1,3-endoglucanase have molecular weight around 48 kD. Antifungal activity of β-1,3-endoglukanase show that it has best inhibition zone on Fusarium solanii at extract from ion exchange chromatography.

scholarly journals Application of High-performance Cation-exchange Chromatography for Characterization of Stinging Insect Venoms.

1983 ◽  
Vol 37b ◽  
pp. 252-254 ◽  
Author(s):  
Roland Einarsson ◽  
Ing-Mari Åstrand ◽  
Per Ålin ◽  
Bengt Mannervik ◽  
Jan-Eric Berg ◽  
...  
Keyword(s):  
Cation Exchange ◽  
High Performance ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Stinging Insect

Preparation and Characterization of NH2-Terminal Fibrinogen Bβ Fragments from N-DSK of Human Fibrinogen

1984 ◽  
Vol 51 (01) ◽  
pp. 016-021 ◽  
Author(s):  
S Birken ◽  
G Agosto ◽  
B Lahiri ◽  
R Canfield
Keyword(s):  
High Performance ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Reverse Phase ◽  
Human Fibrinogen ◽  
Human Volunteers ◽  
Cnbr Cleavage ◽  
Two Peaks

SummaryIn order to investigate the early release of NH2-terminal plasmic fragments from the Bβ chain of fibrinogen, substantial quantities of Bβ 1-42 and Bβ 1-21 are required as immunogens, as radioimmunoassay standards and for infusion into human volunteers to determine the half-lives of these peptides. Towards this end methods that employ selective proteolytic cleavage of these fragments from fibrinogen have been developed. Both the N-DSK fragment, produced by CNBr cleavage of fibrinogen, and Bβ 1-118 were employed as substrates for plasmin with the finding of higher yields from N-DSK. Bβ 1-42 and Bβ 1-21 were purified by gel filtration and ion-exchange chromatography on SP-Sephadex using volatile buffers. When the purified preparation of Bβ 1-42 was chromatographed on reverse-phase high performance liquid chromatography, two peaks of identical amino acid composition were separated, presumably due either to pyroglutamate or to amide differences.

Co-purification of bone resorbing activity and adenylate cyclase stimulating activity from human tumours associated with the humoral hypercalcaemia of malignancy

1987 ◽  
Vol 114 (1) ◽  
pp. 18-26 ◽  
Author(s):  
Chohei Shigeno ◽  
Itsuo Yamamoto ◽  
Shegiharu Dokoh ◽  
Megumu Hino ◽  
Jun Aoki ◽  
...  
Keyword(s):  
Bone Resorption ◽  
High Performance ◽  
Basic Protein ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Protein Factor ◽  
Human Tumours ◽  
Acid Stable ◽  
Bone Resorbing Activity

Abstract. We have partially purified a tumour factor capable of stimulating both bone resorption in vitro and cAMP accumulation in osteoblastic ROS 17/2 cells from three human tumours associated with humoral hypercalcaemia of malignancy. Purification of tumour factor by sequential acid urea extraction, gel filtration and cation-exchange chromatography, reverse-phase high performance liquid chromatography followed by analytical isoelectric focussing provided a basic protein (pI > 9.3) with a molecular weight of approximately 13 000 as a major component of the final preparation which retained both the two bioactivities. Bone resorbing activity and cAMP-increasing activity in purified factor correlated with each other. cAMP-increasing activity of the factor was heat- and acid-stable, but sensitive to alkaline ambient pH. Treatment with trypsin destroyed cAMP-increasing activity of the factor. Synthetic parathyroid hormone (PTH) antagonist, human PTH-(3– 34) completely inhibited the cAMP-increasing activity of the factor. The results suggest that this protein factor, having its effects on both osteoclastic and osteoblastic functions, may be involved in development of enhanced bone resorption in some patients with humoral hypercalcaemia of malignancy.

Preparation and Characterization of the High Molecular Weight [3H]Hyaluronic Acid

1992 ◽  
Vol 57 (10) ◽  
pp. 2151-2156 ◽  
Author(s):  
Peter Chabreček ◽  
Ladislav Šoltés ◽  
Hynek Hradec ◽  
Jiří Filip ◽  
Eduard Orviský
Keyword(s):  
Molecular Weight ◽  
Hyaluronic Acid ◽  
High Molecular Weight ◽  
Methyl Bromide ◽  
High Performance ◽  
Hydrogen Atoms ◽  
Isolation And Characterization ◽  
Labelling Procedure ◽  
Enzymatic Depolymerization

Two methods for the preparation of high molecular weight [3H]hyaluronic acid were investigated. In the first one, hydrogen atoms in the molecule were replaced by tritium. This isotopic substitution was performed in aqueous solution using Pd/CaCO3 as the catalyst. In the second method, the high molecular weight hyaluronic acid was alkylated with [3H]methyl bromide in liquid ammonia at a temperature of -33.5 °C. High-performance gel permeation chromatographic separation method was used for the isolation and characterization of the high molecular weight [3H]hyaluronic acid. Molecular weight parameters for the labelled biopolymers were Mw = 128 kDa, Mw/Mn = 1.88 (first method) and Mw = 268 kDa, Mw/Mn = 1.55 (second method). The high molecular weight [3H]hyaluronic acid having Mw = 268 kDa was degraded further by specific hyaluronidase. Products of the enzymatic depolymerization were observed to be identical for both, labelled and cold biopolymer. This finding indicates that the described labelling procedure using [3H]methyl bromide does not induce any major structural rearrangements in the molecule.

scholarly journals Enzymatic Synthesis and Characterization of Different Families of Chitooligosaccharides and Their Bioactive Properties

2021 ◽  
Vol 11 (7) ◽  
pp. 3212
Author(s):  
Noa Miguez ◽  
Peter Kidibule ◽  
Paloma Santos-Moriano ◽  
Antonio O. Ballesteros ◽  
Maria Fernandez-Lobato ◽  
...  
Keyword(s):  
High Performance ◽  
Chemical Characterization ◽  
Amperometric Detection ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Enzymatic Production ◽  
Bioactive Properties ◽  
Substrate Properties ◽  
Chitin Hydrolysis

Chitooligosaccharides (COS) are homo- or hetero-oligomers of D-glucosamine (GlcN) and N-acetyl-D-glucosamine (GlcNAc) that can be obtained by chitosan or chitin hydrolysis. Their enzymatic production is preferred over other methodologies (physical, chemical, etc.) due to the mild conditions required, the fewer amounts of waste and its efficiency to control product composition. By properly selecting the enzyme (chitinase, chitosanase or nonspecific enzymes) and the substrate properties (degree of deacetylation, molecular weight, etc.), it is possible to direct the synthesis towards any of the three COS types: fully acetylated (faCOS), partially acetylated (paCOS) and fully deacetylated (fdCOS). In this article, we review the main strategies to steer the COS production towards a specific group. The chemical characterization of COS by advanced techniques, e.g., high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and MALDI-TOF mass spectrometry, is critical for structure–function studies. The scaling of processes to synthesize specific COS mixtures is difficult due to the low solubility of chitin/chitosan, the heterogeneity of the reaction mixtures, and high amounts of salts. Enzyme immobilization can help to minimize such hurdles. The main bioactive properties of COS are herein reviewed. Finally, the anti-inflammatory activity of three COS mixtures was assayed in murine macrophages after stimulation with lipopolysaccharides.

Molecular and biochemical characteristics of inulosucrase InuBK from Alkalihalobacillus krulwichiae JCM 11 691

2021 ◽  
Author(s):  
Ken-ji Yokoi ◽  
Sosyu Tsutsui ◽  
Gen-ya Arakawa ◽  
Masakazu Takaba ◽  
Koichi Fujii ◽  
...  
Keyword(s):  
High Performance ◽  
Culture Medium ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Size Exclusion ◽  
Resonance Spectroscopy ◽  
Gene Encoding ◽  
Reading Frame ◽  
Exclusion Chromatography ◽  
Chain Lengths

Abstract Information about the inulosucrase of non-lactic acid bacteria is scarce. We found a gene encoding inulosucrase (inuBK) in the genome of the gram-positive bacterium Alkalihalobacillus krulwichiae JCM 11691. The inuBK open reading frame encoded a protein comprising 456 amino acids. We expressed His-tagged InuBK in culture medium using a Brevibacillus system. The optimal pH and temperature of purified InuBK were 7.0–9.0 and 50 °C–55 °C, respectively. The findings of high-performance anion-exchange chromatography, nuclear magnetic resonance spectroscopy, and high-performance size-exclusion chromatography with multi-angle laser light scattering showed that the polysaccharide produced by InuBK was an inulin with a molecular weight of 3,806, a polydispersity index (PI) of 1.047, and fructosyl chain lengths with 3–27 degrees of polymerization. The size of InuBK was smaller than commercial inulins, and the PI of the inulin that it produced was lower.

Isolation and characterization of the ethynylestradiol-biodegrading microorganism Fusarium proliferatum strain HNS-1

2002 ◽  
Vol 45 (12) ◽  
pp. 175-179 ◽  
Author(s):  
J.H. Shi ◽  
Y. Suzuki ◽  
B.-D. Lee ◽  
S. Nakai ◽  
M. Hosomi
Keyword(s):  
Culture Medium ◽  
Polar Compounds ◽  
Sole Source ◽  
Order Rate Constant ◽  
Fusarium Proliferatum ◽  
Rrna Gene ◽  
28S Rrna ◽  
Isolation And Characterization ◽  
Phenolic Group

We cultivated hundreds of sediment, soil, and manure samples taken from rivers and farms in a medium containing ethynylestradiol (EE2) as the sole source of carbon, so that microorganisms in the samples would acclimatize to the presence of EE2. Finally, we isolated an EE2-degrading microorganism, designated as strain HNS-1, from a cowshed sample. Based on its partial nucleotide sequence (563 bp) of the 28S rRNA gene, strain HNS-1 was identified as Fusarium proliferatum. Over 15 days, F. proliferatum strain HNS-1 removed 97% of EE2 at an initial concentration of 25 mg.L−1, with a first-order rate constant of 0.6 d−1. Unknown products of EE2 degradation, which may be more polar compounds that have a phenolic group, remained in the culture medium.

scholarly journals The copolymeric structure of pig skin dermatan sulphate. Isolation and characterization of l-idurono-sulphate-containing oligosaccharides from copolymeric chains

1974 ◽  
Vol 143 (2) ◽  
pp. 379-389 ◽  
Author(s):  
Lars-Åke Fransson ◽  
Lars Cöster ◽  
Birgitta Havsmark ◽  
Anders Malmström ◽  
Ingrid Sjöberg
Keyword(s):  
Periodate Oxidation ◽  
Exchange Chromatography ◽  
Dermatan Sulphate ◽  
Chondroitinase Abc ◽  
Pig Skin ◽  
Isolation And Characterization ◽  
Smith Degradation ◽  
Iduronic Acid ◽  
Acidic Conditions

Dermatan sulphate was degraded by testicular hyaluronidase and an oversulphated fraction was isolated by ion-exchange chromatography. This preparation, which contained fairly long segments derived from the non-reducing terminal portion of the molecule, was subjected to periodate oxidation under acidic conditions. The oxidized iduronic acid residues were cleaved by reduction-hydrolysis (Smith-degradation) (Fransson & Carlstedt, 1974) or by alkaline elimination. The oligosaccharides so obtained contained both GlcUA (glucuronic acid) and IdUA-SO4 (sulphated iduronic acid) residues. Copolymeric oligosaccharides obtained after alkaline elimination were cleaved by chondroitinase-AC into disaccharide and higher oligosaccharides. Since the corresponding oligosaccharides obtained by Smith-degradation were unaffected by this enzyme, it was concluded that the carbohydrate sequences were GalNAc-(IdUA-GalNAc)n-GlcUA-GalNAc. The iduronic acid-containing sequences were resistant to digestion with chondroitinase-ABC. It was demonstrated that the presence of unsulphated N-acetylgalactosamine residues in these sequences could be responsible for the observed effect. This information was obtained in an indirect way. Chemically desulphated dermatan sulphate was found to be a poor substrate for the chondroitinase-ABC enzyme. Moreover, digestion with chondroitinase-ABC of chondroitinase-AC-degraded dermatan sulphate released periodate-resistant iduronic acid-containing oligosaccharides. It is concluded that copolymeric sequences of the following structure are present in pig skin dermatan sulphate: [Formula: see text] N-acetylgalactosamine moieties surrounding IdUA-SO4 residues are unsulphated to a large extent.

Sign in / Sign up

Export Citation Format

Share Document