Purification of pea nodule symbiosomes using an aqueous polymer two-phase system

J Chromatogr B Biomed Appl. 1996 May 17;680(1-2):171-81. doi: 10.1016/0378-4347(96)88204-1.

Abstract

Symbiosomes were obtained from mature pea (Pisum sativum cv. Argona) root nodules infected with Rhizobium leguminosarum strain (biov. viciae 3841) and purified using an aqueous polymer two-phase system (APS). The APS consists of a mixture of polymers, usually dextran T500 and poly(ethylene glycol) 3350, prepared as aqueous solutions on a weight per weight basis, where each fraction distributes according to their surface characteristics. Results of ATPase activity, cytochrome c oxidase activity, glucan synthase II activity, NAD(P)H-cytochrome c reductase activity, NO3(-)-sensitive ATPase activity, transport of [14C]malate vs. [14C]glutamate and MAC 57 antigen analysis showed that the APS method provided intact symbiosomes with low bacteroid, plasma membrane, endoplasmic reticulum and/or mitochondria contamination. No complicated equipment is needed and the method was simple and fast, compared with other purification techniques.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Cell Membrane / chemistry
  • Cell Membrane / microbiology
  • Cell Membrane / ultrastructure
  • Centrifugation, Density Gradient
  • Dextrans / chemistry
  • Electron Transport Complex IV / metabolism
  • Immunoblotting
  • Pisum sativum / chemistry
  • Pisum sativum / microbiology*
  • Pisum sativum / ultrastructure
  • Plant Roots / chemistry
  • Plant Roots / microbiology*
  • Plant Roots / ultrastructure
  • Polyethylene Glycols / chemistry
  • Polymers / chemistry
  • Rhizobium leguminosarum / chemistry
  • Rhizobium leguminosarum / isolation & purification*
  • Rhizobium leguminosarum / metabolism
  • Solvents / chemistry
  • Surface Properties
  • Symbiosis*

Substances

  • Dextrans
  • Polymers
  • Solvents
  • Polyethylene Glycols
  • Electron Transport Complex IV
  • Adenosine Triphosphatases