Chromatography of Acidic Polysaccharides on Deae-Cellulose

The chromatography of polysaccharides on resinous ion-exchangers has been reported by Steiner, Neukom, and Deuel (1958). Neukom et al. (1960) later reported the chromatography of a variety of wheat and sugar-beet polysaccharides on DEAEcellulose. Only "neutral" polysaccharides could be eluted at pH 6-8 by increasing buffer and salt concentrations; acid polysaccharides, e.g. pectic acid, were eluted by various strengths of alkali. Neutral polysaccharides could also be hromatographed at a somewhat alkaline pH by working with borate systems, which give charged complexes with many carbohydrates.

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Chromatographic resolution of racemates on natural optically active ion exchangers IV. Influence of the solvent on the optical resolution of racemic bases on sunflower pectic acid

Journal of Chromatography A ◽

10.1016/0021-9673(72)80023-2 ◽

1972 ◽

Vol 72 (1) ◽

pp. 192-197 ◽

Cited By ~ 9

Author(s):

Mariana Il. Popova ◽

Christo G. Kratchanov

Keyword(s):

Optical Resolution ◽

Optically Active ◽

Ion Exchangers ◽

Pectic Acid ◽

Chromatographic Resolution

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Physiological and Proteomic Analysis of Different Molecular Mechanisms of Sugar Beet Response to Acidic and Alkaline pH Environment

Frontiers in Plant Science ◽

10.3389/fpls.2021.682799 ◽

2021 ◽

Vol 12 ◽

Author(s):

Gui Geng ◽

Gang Wang ◽

Piergiorgio Stevanato ◽

Chunhua Lv ◽

Qiuhong Wang ◽

...

Keyword(s):

Sugar Beet ◽

Soil Ph ◽

Proteomic Analysis ◽

Molecular Mechanisms ◽

Soluble Sugar ◽

Acid Stress ◽

Pcr Analysis ◽

Alkaline Ph ◽

Growth Status ◽

Leaves And Roots

Soil pH is a major constraint to crop plant growth and production. Limited data are available on sugar beet growth status under different pH conditions. In this study, we analyzed the growth status and phenotype of sugar beet under pH 5, pH 7.5, and pH 9.5. It was found that the growth of sugar beet was best at pH 9.5 and worst at pH 5. The activities of superoxide dismutase (SOD) and peroxidase (POD) in leaves and roots increased as pH decreased from 9.5 to 5. Moreover, compared with pH 9.5, the levels of soluble sugar and proline in leaves increased significantly at pH 5. To explore the mechanisms of sugar beet response to different soil pH environments, we hypothesized that proteins play an important role in plant response to acidic and alkaline pH environment. Thus, the proteome changes in sugar beet modulated by pH treatment were accessed by TMT-based quantitative proteomic analysis. A total of three groups of differentially expressed proteins (DEPs) (pH 5 vs. pH 7.5, pH 9.5 vs. pH7.5 and pH 5 vs. pH 9.5) were identified in the leaves and roots of sugar beet. Several key proteins related to the difference of sugar beet response to acid (pH 5) and alkaline (pH 9.5) and involved in response to acid stress were detected and discussed. Moreover, based on proteomics results, QRT-PCR analysis confirmed that expression levels of three N transporters (NTR1, NRT2.1, and NRT2.5) in roots were relatively high under alkaline conditions (pH 9.5) compared with pH 5 or pH 7.5. The total nitrogen content of pH 9.5 in sugar beet was significantly higher than that of pH 7.5 and pH 5. These studies increase our understanding of the molecular mechanism of sugar beet response to different pH environments.

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Production and properties of polygalacturonate lyase by an alkalophilic microorganism Bacillus sp. RK9

Canadian Journal of Microbiology ◽

10.1139/m78-190 ◽

1978 ◽

Vol 24 (10) ◽

pp. 1164-1172 ◽

Cited By ~ 33

Author(s):

Catherine T. Kelly ◽

William M. Fogarty

Keyword(s):

Ethylenediaminetetraacetic Acid ◽

Divalent Cations ◽

Deae Cellulose ◽

Exchange Chromatography ◽

Bacillus Sp ◽

Pectic Acid ◽

Ph Optimum ◽

Random Fashion ◽

Initial Ph ◽

Polygalacturonate Lyase

Bacillus sp. RK9 was isolated from soil and produced a constitutive polygalacturonate lyase. Production of the enzyme required the presence of complex nitrogen (peptone and yeast extract). Highest activity was obtained with an initial pH of 9.7. The organism was alkalophilic. No growth occurred below pH 7.5. The enzyme was purified by salt precipitation and diethylaminoethyl (DEAE) cellulose ion-exchange chromatography. The pH optimum for activity was 10.0 in 0.01 M glycine–NaOH buffer. Calcium alone, of divalent cations, activated the enzyme by 2.9-fold. Complete inhibition of enzyme activity wasachieved by 1 mM ethylenediaminetetraacetic acid (EDTA). Hydrolysis of substrate occurred in a random fashion and the enzyme was 50% more active towards acid soluble pectic acid (ASPA) than towards sodium polypectate.

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Fasciaton of Pectate Elementary Fibrils

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100070746 ◽

1973 ◽

Vol 31 ◽

pp. 60-61

Author(s):

George F. Leeper

Keyword(s):

Acid Treatment ◽

Carbon Film ◽

Final Concentration ◽

Cellulose Microfibrils ◽

Pectic Acid ◽

Elementary Fibrils

Polysaccharide elementary fibrils are usually fasciated into microfibrils of from one hundred to a few hundred Angstroms wide. Cellulose microfibrils when subjected to acid treatment dissociate into component elementary fibrils. For pectic acid it was observed that variations in pH could cause a change in the fasciation of the elementary fibrils.Solutions of purified pectic acid and sodium phosphotungtate were adjusted to various pH levels with NaOH or HCl and diluted to give a final concentration of 0.5 and 1% for the polysaccharide and negative stains respectively. Micrographs were made of the samples after drying on a carbon film covered grid. The average number of elementary fibrils was determined by counting the number of elementary fibrils in each fascicle intersected by lines drawn across the micrograph.

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Three-Dimensional Structure of Cloned T3 Connector Protein at 1.6nm Resolution

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100180161 ◽

1990 ◽

Vol 48 (1) ◽

pp. 282-283

Author(s):

José L. Carrascosa ◽

José M. Valpuesta ◽

Hisao Fujisawa

Keyword(s):

Dna Sequences ◽

Expression Vector ◽

Three Dimensional ◽

Deae Cellulose ◽

Dna Packaging ◽

Dimensional Structure ◽

Two Dimensional ◽

Freezing And Thawing ◽

Three Dimensional Structure ◽

Dimensional Reconstruction

The head to tail connector of bacteriophages plays a fundamental role in the assembly of viral heads and DNA packaging. In spite of the absence of sequence homology, the structure of connectors from different viruses (T4, Ø29, T3, P22, etc) share common morphological features, that are most clearly revealed in their three-dimensional structure. We have studied the three-dimensional reconstruction of the connector protein from phage T3 (gp 8) from tilted view of two dimensional crystals obtained from this protein after cloning and purification.DNA sequences including gene 8 from phage T3 were cloned, into Bam Hl-Eco Rl sites down stream of lambda promotor PL, in the expression vector pNT45 under the control of cI857. E R204 (pNT89) cells were incubated at 42°C for 2h, harvested and resuspended in 20 mM Tris HC1 (pH 7.4), 7mM 2 mercaptoethanol, ImM EDTA. The cells were lysed by freezing and thawing in the presence of lysozyme (lmg/ml) and ligthly sonicated. The low speed supernatant was precipitated by ammonium sulfate (60% saturated) and dissolved in the original buffer to be subjected to gel nitration through Sepharose 6B, followed by phosphocellulose colum (Pll) and DEAE cellulose colum (DE52). Purified gp8 appeared at 0.3M NaCl and formed crystals when its concentration increased above 1.5 mg/ml.

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