http://orcid.org/0000-0001-5018-0933
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ABSTRACT
Date fruits are well known to be very nutritious. Nevertheless, the protein contents of the fruit, particularly the seed and flesh, are still understudied, largely due to their difficult physical characteristics. This study was conducted to compare three different protein extraction methods which were the trichloroacetic acid (TCA)-acetone (TCA-A), phenol (Phe), and TCA-acetone-phenol (TCA-A-Phe), and to perform proteomic analysis on date palm seed and flesh. Phe extraction method showed the highest protein yields for both seed (8.26 mg/g) and flesh (1.57 mg/g). Through sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Phe, and TCA-A-Phe extraction methods were shown to be efficient in removing interfering compounds and gave well-resolved bands over a wide range of molecular weights. Following liquid chromatography–tandem mass spectrometry analysis, about 50–64% of extracted proteins were identified with known functions including those involved in glycolysis, Krebs cycle, defense, and storage. Phe protein extraction method was proven to be the optimal method for date flesh and seed.
Abbreviations::
- TCA: trichloroacetic acid
- Phe: phenol
- A: acetone
- SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- LC-MS/MS: liquid chromatography–tandem mass spectrometry
- Tris: tris(hydroxymethyl)aminomethane
- EDTA: ethylenediaminetetraacetic acid disodium salt
- KCl: potassium chloride
- CHAPS: 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate hydrate
- BSA: bovine serum albumin
- HCl: hydrochloric acid
- DTT: 1,4-dithiothreitol
- NaOH: sodium hydroxide
- IAM: α-iodoacetamide
- MW: molecular weight