Identification of extensin protein associated with sugar beet pectin.
Several studies have suggested that the emulsification properties associated with pectin obtained from sugar beet (Beta vulgaris) are due to the presence of a protein-pectin complex. Nevertheless, the identity of the protein has remained elusive. Pectin, extracted from sugar beet pulp by microwave-assisted extraction, and a commercial sample were both subjected to protease digestion with trypsin. The resulting peptides were separated from the pectin solution by ultrafiltration using a 3 kDa molecular weight cutoff (MWCO) membrane and analyzed using matrix-assisted laser desorption ionization with tandem time-of-flight mass spectrometry. The partial sequences derived from the mass spectrometry analyses of the resulting tryptic peptides are found to be highly consistent with extensin protein matched from the B. vulgaris Genetic Index database and also correspond to previously reported extensin peptides found in sugar beet cell suspension cultures. Further attempts were made to disassociate the protein from pectin using 1 M NaCl and a 100 kDa MWCO membrane; however, no peptides were observed following trypsin digestion of the permeate solution. This evidence suggests the existence of a complex between the pectin and extensin that is not due to ionic interactions. Trypsin digestion of commercial sugar beet pectin also produced the peptide profile observed with the microwave-assisted extracted pectin sample. Atomic force microscopy established that the number of rod-like elements decreased following protease treatment compared to the untreated sample.
Nuñez
Alberto
A
Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, 600 East Mermaid Lane, Wyndmoor, Pennsylvania 19038, USA. alberto.nunez@ars.usda.gov
Fishman
Marshall L
ML
Fortis
Laurie L
LL
Cooke
Peter H
PH
Hotchkiss
Arland T
AT
D000595:Amino Acid Sequence; D027461:Beta vulgaris / Q000737:chemistry*; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry; D018625:Microscopy, Atomic Force; D008969:Molecular Sequence Data; D010368:Pectins / Q000737:chemistry* / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010449:Peptide Mapping; D010940:Plant Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D018517:Plant Roots / Q000737:chemistry*; D014357:Trypsin / Q000378:metabolism
D006023:Glycoproteins; D010446:Peptide Fragments; D010940:Plant Proteins; C005488:extensin protein, plant; D010368:Pectins; D014357:Trypsin