摘要
This paper studies the rapid identification of heavy metal-binding peptides (phytochelatin) by taking Water Hyacinth as a model plant. Plants were cultured in water containing 2 μg/ml Cd2+ for 13 days. The Sephadex G-50 chromatography of root extract under low salt concentration (0. 01 mol/L PBS) gave a Cd-binding peak with MW of 10 ,000 determined by SEC HPLC. After oxidation with performic acid, its SEC HPLC molecular weight decreased to below 1300 and the reverse phase HPLC showed one peptide peak, whose amino acid composition is the same as that of the sample never undertaking oxidation, and (Glu/Gln):Cys:Gly=2:2:1. According to the general structure of phytochelatin (γ-Glu-Cys)n-Gly, n is 2 in this case. The protocol including the sequential steps of Sephadex G-50 chromatography→performic acid oxidation→reverse phase HPLC→amino acid analysis is a rapid and effective method to identify the existence of phytochelatin and determine its values of n.
This paper studies the rapid identification of heavy metal-binding peptides (phytochelatin) by taking Water Hyacinth as a model plant. Plants were cultured in water containing 2 μg/ml Cd2+ for 13 days. The Sephadex G-50 chromatography of root extract under low salt concentration (0. 01 mol/L PBS) gave a Cd-binding peak with MW of 10 ,000 determined by SEC HPLC. After oxidation with performic acid, its SEC HPLC molecular weight decreased to below 1300 and the reverse phase HPLC showed one peptide peak, whose amino acid composition is the same as that of the sample never undertaking oxidation, and (Glu/Gln):Cys:Gly=2:2:1. According to the general structure of phytochelatin (γ-Glu-Cys)n-Gly, n is 2 in this case. The protocol including the sequential steps of Sephadex G-50 chromatography→performic acid oxidation→reverse phase HPLC→amino acid analysis is a rapid and effective method to identify the existence of phytochelatin and determine its values of n.
