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Plant Foods for Human Nutrition

, Volume 67, Issue 3, pp 247–255 | Cite as

Solubilization, Fractionation, and Electrophoretic Characterization of Inca Peanut (Plukenetia volubilis L.) Proteins

  • Shridhar K. Sathe
  • Harshal H. Kshirsagar
  • Girdhari M. Sharma
Original Paper

Abstract

Effects of different solvents, ionic strength, and pH on Inca peanut seed protein solubility were assessed by quantitatively analyzing solubilized proteins using Lowry and Bradford methods. Soluble proteins were fractionated using Osborne procedure and the polypeptide composition of solubilized proteins was determined by one dimensional 25 % monomer acrylamide linear gradient SDS-PAGE. Osborne protein fractions were analyzed by the 2D gel electrophoresis. Total seed proteins were efficiently solubilized by 2 M NaCl among the tested solvents. The soluble seed proteins registered a minimum solubility at pH ~4.0. Osborne protein fractions, albumins, globulins, prolamins, and glutelins accounted for 43.7, 27.3, 3.0, and 31.9 %, respectively, of the total aqueous soluble proteins. Soluble seed flour proteins are mainly composed of polypeptides in the MW range of 6–70 kDa of which the predominant polypeptides were in the 20–40 kDa range. Prolamin fraction was mainly composed of four polypeptides (MW < 15 kDa). Glycoprotein staining indicated 32–35 and <14 kDa peptides to be positive.

Keywords

Inca peanut Protein Solubility Protein fractions Electrophoresis 

Abbreviations

β-ME

β-mercaptoethanol

CBBR

Coomassie brilliant blue R

CHAPS

3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate

DTT

Dithiothreitol

EtOH

Ethanol

IEF

Isoelectric focusing

kDa

Kilodalton

LSD

Least significant difference

MeOH

Methanol

MW

Molecular weight

PAGE

Polyacrylamide gel electrophoresis

pI

Isoelectric pH

RT

Room temperature

SDS

Sodium dodecyl sulfate

Tris

2-Amino-2-hydroxymethyl-propane-1,3-diol

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Copyright information

© Springer Science+Business Media, Inc. 2012

Authors and Affiliations

  • Shridhar K. Sathe
    • 1
  • Harshal H. Kshirsagar
    • 1
    • 2
  • Girdhari M. Sharma
    • 1
    • 3
  1. 1.Department of Nutrition, Food & Exercise Sciences, College of Human SciencesThe Florida State UniversityTallahasseeUSA
  2. 2.Roquette America Inc.GenevaUSA
  3. 3.Department of Health and Human Services, Public Health Service, Food and Drug Administration, Office of Applied Research and Safety AssessmentCenter for Food Safety and Applied Nutrition, U.S. FDALaurelUSA

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