紫羊茅根中铜结合肽的分离和纯化

ISOLATION AND PURIFICATION OF METAL BINDING PEPTIDE FROM COPPER TREATED FESTUCA RUBRA cv. MERLIN ROOTS

利用１次ＳｅｐｈａｄｅｘＧ５０凝胶过滤和２次ＱＡＥＳｅｐｈａｄｅｘＡ２５离子交换层析的方法，从最后６ｄ用ＣｕＣｌ２处理（ρ＝２０ｍｇ／Ｌ）、２０ｄ龄的紫羊茅（Ｆｅｓｔｕｃａｒｕｂｒａｃｖ．Ｍｅｒｌｉｎ）根中分离纯化了１个铜结合肽（ＣｕＢＰ２ａ），该肽在ＳｕｐｅｒｄｅｘＰｅｐｔｉｄｅＨＲ１０／３０预装柱的ＦＰＬＣ凝胶过滤中在λ２５４ｎｍ处只出现一个峰，经分子量标准曲线估计，其Ｍｒ约为１４００，在ＤｅｌｔａＰａｋＣ１８的反相ＨＰＬＣ中，在λ２５４ｎｍ处也只有１个紫外吸收峰值，这都说明该肽已经达到了较高的的纯度．通过光谱分析，发现该肽的最大紫外吸收值在２４８．１ｎｍ处．对Ｃｕ结合物在植物体内的存在状态进行了讨论．

Through a gel filtration chromatography on Sephadex G 50 and twice on FPLC ion exchange chromatography on QAE Sephadex A 25, a metal binding peptide (CuBP 2a) was purified from 20 day old F. rubra roots treated by copper (supplied with CuCl 2) at the concentration of 20 mg/L during the last 6 days. CuBP 2a showed a single peak at 254 nm and was about 1 400 doltons after the calibration by standard molecular weight markers in FPLC gel filtration on pre packed Superdex Peptide HR 10/30 column. Its purity was judged by the single peak at 254 nm in the elution profile of reversed phase HPLC on Delta Pak C18. By UV absorbance spectrum analysis, the peptide showed the maximal absorbance at 248.1nm. The metal binding peptides in plant cells are also disscussed.