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Hemagglutinating activity and corresponding putative sequence identity from Curcuma aromatica rhizome
Author(s) -
Tiptara Ponpimol,
Petsom Amorn,
Roengsumran Sophon,
Sangvanich Polkit
Publication year - 2008
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.3184
Subject(s) - chromatography , chemistry , size exclusion chromatography , ammonium sulfate precipitation , tandem mass spectrometry , rhizome , peptide sequence , gel electrophoresis , biology , biochemistry , mass spectrometry , botany , enzyme , gene
BACKGROUND: Curcuma aromatica is a medicinal plant belonging to the Zingiberaceae family with an incomplete genome sequence. It has been reported that extract from the rhizome of this plant contains haemagglutinating activity. In this study the profile of fractions containing hemagglutinating activity is described. RESULTS: Following extraction with saline buffer, the protein solution was fractionated by ammonium sulfate precipitation. Ion‐exchange chromatography was completed on fast‐flow SP‐Sepharose, as well as gel filtration chromatography on Superdex 75. The active fractions were then separated by one‐dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis and labeled proteins were digested with trypsin. The digest bands were analyzed by reversed‐phase liquid chromatography—tandem mass spectrometry. Inferred peptide sequences were used in Mascot searching and mass spectrometry‐driven BLAST (MS‐BLAST) homology searches allowed the recognition of related proteins in other species of Viridiplantae. Six putative proteins from nine bands showed similarity with lectin sequences. CONCLUSION: This study reports the identification of six lectins from the Curcuma aromatica rhizome achieved by mass spectrometry using MS‐BLAST algorithms to search for homology between de novo determined peptide sequences and protein sequences available in sequence databases. Copyright © 2008 Society of Chemical Industry