Complete solubilization of pectins from cotton suspension culture cell walls with retention of most structural features
- Oklahoma State Univ., Stillwater (United States)
- Oklahoma State Univ., Stillwater (United States) Univ. of Georgia, Athens (United States)
- Oklahoma State Univ., Stillwater (United States) Univ. of Cincinnati, OH (United States)
Cotton suspension culture cell walls contain four major pectin substructures: (1) rhamnogalacturonan I (RGI), (2) rhamnogalacturonan II, (3) highly methyl esterified homogalacturonan, and (4) homogalacturonan of low degree of methyl esterification. Methods are described to solubilize and isolate each of the four major substructures from the cell walls in high yields. RGII is completely solubilized by an easily purified endopolygalacturonase (EPG). Around 40% of RGI can be solubilized by the sequential action of this EPG and a commercially available cellulase. Almost all of the RGI along with xyloglucan can be solubilized after the EPG treatment using strong alkali. Highly methyl esterified homogalacturonan (degree of methyl esterification [approximately]40%) is solubilized into water after HF treatment of the untreated walls, at [minus]23[degrees], and homogalacturonans with a very low degree of methyl esterification ([approximately]10%) can subsequently be solubilized into 500 mM imidazole buffer. the highly methyl esterified homogalacturonan co-solubilizes with the RGI during several extraction procedures and co-chromatographs with it, indicating that they are covalently attached to each other in the cell walls. Little of the RGI is solubilized from cotton walls by EPG digestion without a subsequent treatment that co-solubilizes the xyloglucan or degrades the xyloglucan, indicating crosslinks exist between the RGI and much of the xyloglucan.
- OSTI ID:
- 6282058
- Report Number(s):
- CONF-9307119--
- Journal Information:
- Plant Physiology; (United States), Journal Name: Plant Physiology; (United States) Vol. 102:1; ISSN 0032-0889; ISSN PLPHAY
- Country of Publication:
- United States
- Language:
- English
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