The TiO2-densified cellulose composite beads were activated by epichlorohydrin and coupled with diethylamine, to function as an anion exchanger for expanded bed chromatography. The adsorbent exhibited a favorable perf...The TiO2-densified cellulose composite beads were activated by epichlorohydrin and coupled with diethylamine, to function as an anion exchanger for expanded bed chromatography. The adsorbent exhibited a favorable performance of expanded bed adsorption for proteins, and therefore was applied to the expanded bed recovery of D-2-chloropropionic acid dehalogenase directly from the unclarified homogenate of Pseudomonas sp. NT21. The binding capacity of the dehalogenase was found to be 8.54U·ml^-1 adsorbent, and two active peaks were eluted respectively at 0.15mol·L^-1 and 0.3mol·L^-1 （NH4）2SO4. The result indicated that the overall enzyme yield was 68%, with a purification factor of 22. In comparison to other recovery processes, the yield of the expanded bed process rises at least 70%, simultaneously saving a great deal of operation time and costs.展开更多
清蛋白是扁桃种仁中主要蛋白质,为揭示扁桃仁主要蛋白质组成及结构特性,以新疆莎车1号(SC-1)扁桃仁为研究对象,对其种仁中2S-清蛋白进行分离纯化和结构特性研究,即经有机溶剂多次沉淀提取,获得扁桃仁清蛋白的粗提物,再经Q Sepharose FF...清蛋白是扁桃种仁中主要蛋白质,为揭示扁桃仁主要蛋白质组成及结构特性,以新疆莎车1号(SC-1)扁桃仁为研究对象,对其种仁中2S-清蛋白进行分离纯化和结构特性研究,即经有机溶剂多次沉淀提取,获得扁桃仁清蛋白的粗提物,再经Q Sepharose FF分离纯化,获得扁桃仁2S-清蛋白。经高效液相色谱-质谱联用分析表明,纯化后的扁桃仁2S-清蛋白准确分子质量为31.40 k D,其在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结果中显示为18.0 k D和19.5 k D 2个条带,表明其由两个不同的亚基通过二硫键结合而成。扁桃仁2S-清蛋白中谷氨酸(Glu)、天冬氨酸(Asp)和精氨酸(Arg)的含量最高;其二级结构中主要为β-折叠,占65.9%;扁桃仁2S-清蛋白羰基含量为4.45 nmol/mg、活性巯基含量为39.95?μmol/g、总巯基含量为57.11?μmol/g、表面疏水性为23.45?μg;变性温度为43.3℃。研究结果可为新疆扁桃仁的开发利用提供理论依据。展开更多
基金the Natural Science Foundation of Ningbo （No.2005A610018）.ACKN0WLEDGEMENTS The authors would like to thank Mr. Qi Nan for taking part in this study, and for financial support by the Natural Science Foundation of Ningbo. Discussion with Dr. Lin Dong-Qiang has been helpful and is appreciated.
文摘The TiO2-densified cellulose composite beads were activated by epichlorohydrin and coupled with diethylamine, to function as an anion exchanger for expanded bed chromatography. The adsorbent exhibited a favorable performance of expanded bed adsorption for proteins, and therefore was applied to the expanded bed recovery of D-2-chloropropionic acid dehalogenase directly from the unclarified homogenate of Pseudomonas sp. NT21. The binding capacity of the dehalogenase was found to be 8.54U·ml^-1 adsorbent, and two active peaks were eluted respectively at 0.15mol·L^-1 and 0.3mol·L^-1 （NH4）2SO4. The result indicated that the overall enzyme yield was 68%, with a purification factor of 22. In comparison to other recovery processes, the yield of the expanded bed process rises at least 70%, simultaneously saving a great deal of operation time and costs.
文摘清蛋白是扁桃种仁中主要蛋白质,为揭示扁桃仁主要蛋白质组成及结构特性,以新疆莎车1号(SC-1)扁桃仁为研究对象,对其种仁中2S-清蛋白进行分离纯化和结构特性研究,即经有机溶剂多次沉淀提取,获得扁桃仁清蛋白的粗提物,再经Q Sepharose FF分离纯化,获得扁桃仁2S-清蛋白。经高效液相色谱-质谱联用分析表明,纯化后的扁桃仁2S-清蛋白准确分子质量为31.40 k D,其在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结果中显示为18.0 k D和19.5 k D 2个条带,表明其由两个不同的亚基通过二硫键结合而成。扁桃仁2S-清蛋白中谷氨酸(Glu)、天冬氨酸(Asp)和精氨酸(Arg)的含量最高;其二级结构中主要为β-折叠,占65.9%;扁桃仁2S-清蛋白羰基含量为4.45 nmol/mg、活性巯基含量为39.95?μmol/g、总巯基含量为57.11?μmol/g、表面疏水性为23.45?μg;变性温度为43.3℃。研究结果可为新疆扁桃仁的开发利用提供理论依据。
