Mannans chromatography

Bedair, M., and El Rassl, Z. (2004). Affinity chromatography with monolithic capillary columns I. Polymethacrylate monoliths with Immobilized mannan for the separation of mannose-binding proteins by capillary electrochromatography and nano-scale liquid chromatography. /. Chromatogr. A 1044, 177-186. 

MBP is purified from rabbit serum using affinity chromatography on immobilized mannan (3). The resulting dialyzed MBP may be used directly to couple to activated Sepharose 4B. 

The use of quantitative, paper chromatography is discussed in this Section, since one of the most important factors in its application to wood-cellulose analysis is that it permits a satisfactory determination of mannan. Reasonably reliable methods for the determination of xylan have been available for some time, but mannan determinations, for the reasons discussed above, have been less satisfactory. The fact that (on chromatograms) xylose, as well as other carbohydrates, can be determined simultaneously with mannose is an added attraction in the use of this technique. 

K17. Krol, G. J., Banovsky, J. M., Mannan, C. A., Pickering, R. E., and Kho, B. T., Trace analysis of the MIF analogue parapeptide in blood plasma by high-performance liquid chromatography and short-wavelength excitation fluorometry. ]. Chromatogr. 163, 383-389 (1979). 

Key words Affinity chromatography, Affinity matrices, Antigen-specific antibodies, Hapten-specific antibodies, Specie-specific antibodies, Cross-reacting immunoglobulin, Sepharose, Affigel, IgM, IgA, Mannan binding protein, Jacalin, Protein A, Protein G, Protein L... 

Some idea of the future trend of methylation analysis can, perhaps, be gained from the work of Perila and Bishop, who have used the method to study oligosaccharides obtained by the enzymic hydrolysis of jack-pine mannan. The oligosaccharides (0.5-2.0 mg.) were methylated by the Kuhn procedure, refluxed with methanolic hydrogen chloride, and the resulting methyl 0-methylglycosides analyzed qualitatively and quantitatively by gas-liquid chromatography. 

First, by using alkali-insoluble glucan rather than cell walls as enzyme inducer, enzyme purification was greatly facilitated since yeast mannan (which is not attacked by B. circulans) does not accumulate as a very viscous component in concentrated crude enzyme solutions to be applied to chromatography columns. Yeast glucan is at least as effective as an enzyme inducer as are yeast cell walls. 

P-Mannan is water-insoluble and solubilizing it requires very strong alkali, cuprammonium or derivatization. X-ray and electron diffraction show that mannan has a ribbon-like conformation, similar to cellulose, with the 2-OH axial instead of equatorial. In seeds (palm, coffee, caraway) a few (<5%) of a-Gal groups are attached to the hydroxymethyl and in seaweeds a small percentage (<5) of Glc residues may occur in the chain. A pure P(l-4) mannan has been extracted from cell walls of the siphonous green alga Codium latum as the methylol mannan with paraformaldehyde dimethylsulphoxide. It was purified by size exclusion chromatography in dimethyl sulphoxide and recovered by the addition of water or methanol and could be re-dissolved in hot dimethyl sulphoxide. Infra-red and n.m.r. spectroscopy and periodate oxidation confirmed the unbranched P(l-4) structure [185]. 

Many species of yeasts are known to produce mannans containing various amounts of phosphate, which can be fractionated by anion-exchange chromatography (1, 2,. Recently, we... 

Figure L Elution profiles of the bulk mannans of three C. albicans strains (Batch I) by DEAE—Sephadex chromatography (A-50, acetate, 4 X 25 cm) using a stepwise elution system consisting of water and NaCl solutions, Ten-fxL aliquots of fractions were assayed for carbohydrate content with phenol-sulfuric acid reagent (13) (A) C. albicans NIH A-207 (A-strain) (B) C. albicans NIH B-792 (B-strain) (C) C. albicans J-1012 (J-strain) (11),...

Proteins of a crude enzyme preparation obtained from the cultivation medium of the basidiomycete Phellinus abietis were separated by gel filtration and ion-exchange chromatography. The preparation contained a minimum of three enzymes capable of splitting a-D-mannosidic bonds a-D-mannosidase, exo-D-mannanase, and e/ido-D-mannanase, which were separated. Some properties of the D-mannanase complex of the crude enzyme preparation, and of a partially purified a-D-mannosidase, were examined. The D-mannanase complex exhibited two pH optima, its temperature optimum being 45 C. The pH optimum of purified a-D-mannosidase was at pH 5.0, the temperature optimum was at 60 °C the enzyme had a relatively high heat stability. The of a-D-mannosidase for 4-nitrophenyl a-D-mannopyranoside was 1.5xl0 M. Pure a-D-mannosidase did not split D-mannan. 

The a-D-mannanase activity secreted by an Acinetobacter sp. growing on baker s yeast has been separated into a number of components by ion-exchange chromatography one of them (an exo-a-l,2-D-mannanase) hydrolysed Saccharomyces cerevisiae D-mannan and oligosaccharides derived from yeast D-mannan. ... 

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