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An h.p.l.c. method for determining chain-length distribution in some glycogens.

N W Cheetham1, N Hansawek, P Saecou

  • 1School of Chemistry, University of New South Wales, Kensington, Sydney, Australia.

Carbohydrate Research
|August 12, 1991
PubMed
Summary

Researchers analyzed glycogen chain lengths from human, oyster, and bacterial sources using Pseudomonas amylodermosa isoamylase. The resulting chain length "fingerprints" are unique to each glycogen source, aiding in comparison and isolation monitoring.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Microbiology

Background:

  • Glycogen, a glucose polymer, serves as an energy reserve in various organisms.
  • Understanding glycogen structure, specifically chain length distribution, is crucial for biochemical and physiological studies.
  • Current methods for analyzing glycogen chain lengths can be complex and require refinement.

Purpose of the Study:

  • To characterize the chain length distribution of glycogen from different sources.
  • To evaluate the utility of high-performance liquid chromatography (HPLC) for glycogen analysis.
  • To establish a method for distinguishing between various glycogen types based on their structural "fingerprints".

Main Methods:

  • Glycogen samples from human, oyster, and Streptococcus mitis were debranched using Pseudomonas amylodermosa isoamylase.

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  • Chain length distribution was analyzed using high-performance liquid chromatography (HPLC) on reversed-phase columns with water as the eluent.
  • Oligosaccharides were detected up to a degree of polymerization (d.p.) of 26, with quantitative data obtained for d.p. 3-18.

    • Main Results:

    • Two HPLC columns were required for the quantitative analysis of the complete range of oligosaccharides.
    • The chain length distribution patterns, or "fingerprints," varied significantly among the different glycogen sources.
    • The method provided detailed quantitative data on oligosaccharide chain lengths.

    Conclusions:

    • HPLC analysis following enzymatic debranching provides characteristic chain length "fingerprints" for different glycogen sources.
    • This technique is valuable for comparing glycogen structures and for monitoring glycogen isolation procedures.
    • The findings contribute to a better understanding of glycogen heterogeneity across species.