海藻中多肽(PC_2～PC_4)的测定及对Zn^(2+)诱导响应

Determination and Response of polypeptides( PC_2～PC_4) in algae to Zn^(2+) inducing

本研究以海带为试验载体,建立了海藻中多肽(γ-Glu-Cys)n-Gly的分离、提取技术,并建立了反相C18色谱柱分离-高效液相色谱-荧光法测定海带中多肽的含量;研究中利用不同浓度的Zn2+胁迫培养藻体,考察了海带对Zn2+的富集能力,探索了藻体中H-(γ-Glu-Cys)2-Gly-OH(PC2),H-(γ-Glu-Cys)3-Gly-OH(PC3),H-(γ-Glu-Cys)4-Gly-OH(PC4)3种肽对Zn2+的诱导响应。结果显示,高效液相色谱-荧光法测定多肽化合物,PC2、PC3和PC4分离效果较好,3种化合物出峰时间主要集中在16.6~20.2 min间,方法线性相关性良好(r>0.9995),检出限较低,重线性较好(RSD<2%),该方法实现了多肽化合物快速、准确定量分析;海带对Zn2+具有较强的富集能力,随着培养时间的增长及金属浓度的增加,藻体内Zn2+含量增加明显;藻体中PC2、PC3和PC4随着Zn2+诱导时间的增长其浓度稍微增加,并与藻体内Zn2+的含量呈现正相关。

In this study, the separation and purification method of polypeptide （γ-Glu-Cys） n-Gly in algae was devel- oped. The polypeptides concentration in kelp was determined by reversion phase C18-high performance liquid chroma- tography with fluorescence detector. The kelp was induced by Zn2＋ under different concentrations, the enrichment a- bility for Zn2 ＋ and the variation trend of H- （ γ-Glu-Cys ） 2-Gly-OH （ PC2 ）, H- （ γ-Glu-Cys ） 3-Gly-OH （ PC3 ） and H- （γ-Glu-Cys） 4-Gly-OH（ PC4 ） were also studied. Results showed that the three compounds were well separated with retention times between 16.6 min and 20.2 min. The high performance liquid chromatography method had good linear correlation coefficient（r 〉0. 9995）, low limit of detection and good reproducibility（RSD 〈2% ）. The kelp had a stronger enrichment ability of Zn2 ＋ . The content of Zn2 ＋ in algae increased obviously with metal concentration and in- ducing time increasing. The concentration of PC2 ,PC3 and PC4 in algae increased slightly with metal inducing time in- creasing, and the content of polypeptide presented a positive correlation with Zn2 ＋ concentration.