Cell wall microcapsules with different porosity from suspension cultured Chenopodium album

Abstract

The primary cell wall of suspension cultured higher plant tissues may be used for size fractionation of macromolecules of different ranges of molecular mass by permeation chromatography, as the size limits of cell wall penetration may be adjusted by controlled decay of polygalacturonan. If care is taken to prevent pectin decay by β-elimination during the preparation of the vesicular packing material (VP) from suspension grown cell clusters of Chenopodium album (low temperature, pectin-preserving proteolysis buffer and washing solutions), extraction with ethanol and deproteinization with a pancreatic enzyme mixture did not significantly increase the size limit of permeation (SLP) of the native cell wall. However, the SLP was significantly increased after long term washing at room temperature or heat treatment with phosphate buffer at pH 7. If the VP was incubated in 2% Na2CO3 solution at room temperature, its SLP increased significantly within the first 30 min but remained stable on further incubation in the alkaline solution. The extent of the porosity change of the cell wall occurring in Na2CO3 solution may be varied by choice of the incubation temperature. Permeation chromatography using the variants of the VP with increased porosity enables separation between proteins with large peak separation in the range of middle (50–100 kD) and large (300 kD) molecular mass.