Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu)

Int J Biol Macromol. 2017 Jan;94(Pt A):161-169. doi: 10.1016/j.ijbiomac.2016.09.100. Epub 2016 Sep 29.

Abstract

The aim of this study was to characterize a polysaccharide found in citrus peels with an anti-metastatic property. CPE-II was purified by the pectinase digestion of citrus peels. During in vivo lung metastasis of Colon26-M3.1, administration of 10μg of CPE-II per mouse showed 81.3% inhibition of metastasis. CPE-II consists of 15 different monosaccharides and 22 different glycosyl linkages, characteristic of rhamnogalacturonan II (RG-II). The primary structure was elucidated based on sugar composition, methylation analysis, oligosaccharide analysis, and sequencing using GC, GC-MS, LC-MS, and ESI-MS/MS analyses. Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. The molecular weight of CPE-II was observed to decrease from 9 to 5kDa at a pH value of <2.0, as observed by HPSEC. Thus, we propose that the anti-metastatic CPE-II is primarily present as an RG-II dimer.

Keywords: Anti-metastatic activity; Citrus peels; Pectin; Polysaccharide; Rhamnogalacturonan II.

MeSH terms

  • Animals
  • Antineoplastic Agents, Phytogenic / chemistry
  • Antineoplastic Agents, Phytogenic / isolation & purification
  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Carbohydrate Conformation
  • Cell Line, Tumor
  • Citrus / chemistry
  • Colonic Neoplasms / drug therapy*
  • Colonic Neoplasms / pathology
  • Female
  • Fruit / chemistry
  • Lung Neoplasms / prevention & control*
  • Lung Neoplasms / secondary
  • Mice, Inbred BALB C
  • Neoplasm Transplantation
  • Pectins / chemistry
  • Pectins / isolation & purification
  • Pectins / pharmacology*
  • Polygalacturonase / chemistry

Substances

  • Antineoplastic Agents, Phytogenic
  • rhamnogalacturonan II
  • Pectins
  • Polygalacturonase