Application and comparison of four selected procedures for the isolation of cell-wall material from the skin of grapes cv. Monastrell.
In order to choose an appropriate cell-wall material (CWM) isolation procedure in grapes cv. Monastrell, four different standard procedures have been tested, and a comparison made of the amount of cell-wall material obtained, its composition and morphology. The CWM was isolated as the 70% ethanol insoluble residue (de Vries method), as the absolute ethanol insoluble residue filtered sequentially through nylon mesh (Nunan method), as the insoluble residue in sodium deoxycholate-phenol-acetic acid-water (Selvendran method) and as the N-[2-hydroxyethyl]-piperazine-N'-2-ethanesulfonic acid (HEPES) insoluble residue (Vidal method). All extractions were done in triplicate and the efficiency of the extractive procedure established. Carbohydrates, proteins, and phenolic compounds were analysed, as the main constituents of CWM. The morphology of the isolated CWM was visualized by scanning electron microscopy (SEM). The Selvendran method had the highest efficiency, while the Nunan method had the lower one. Regarding the carbohydrates composition, the four different CWM were rich in uronic acids and glucose, together with varying amounts of arabinose, xylose, mannose and galactose. The Selvendran method had the lower value of total carbohydrates and the CWM shows more plasmatic membrane impurities in SEM images. The chemical results of the Vidal and de Vries methods were quite similar, but the Vidal method was more time consuming than the de Vries method. According to the results, the de Vries method was chosen to produce a representative cell-wall material fraction from Monastrell grapes skin.
Apolinar-Valiente
R
R
Departamento de Tecnología de Alimentos, Nutrición y Bromatología, Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, 30100 Murcia, Spain. tokay04@hotmail.com
Romero-Cascales
I
I
López-Roca
J M
JM
Gómez-Plaza
E
E
Ros-García
J M
JM
D019342:Acetic Acid / Q000737:chemistry; D000107:Acetylation; D002241:Carbohydrates / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D002473:Cell Wall / Q000737:chemistry* / Q000648:ultrastructure; D003840:Deoxycholic Acid / Q000737:chemistry; D000431:Ethanol / Q000737:chemistry; D006531:HEPES / Q000737:chemistry; D008745:Methylation; D010636:Phenols / Q000737:chemistry / Q000302:isolation & purification; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification; D027843:Vitis / Q000737:chemistry*; D014867:Water / Q000737:chemistry
D002241:Carbohydrates; D010636:Phenols; D010940:Plant Proteins; D003840:Deoxycholic Acid; D014867:Water; D000431:Ethanol; D019342:Acetic Acid; D006531:HEPES