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
目的探讨神经元特异性烯醇化酶(Neuron Specific Enolase,简称NSE)、星形胶质源性蛋白(S-100βprotein,简称S-100β)和环氧化酶-2(Cyclooxygenase-2,简称COX-2)在1-溴丙烷大鼠亚急性神经损伤中的变化。方法选取健康雄性Wistar大鼠36只,...目的探讨神经元特异性烯醇化酶(Neuron Specific Enolase,简称NSE)、星形胶质源性蛋白(S-100βprotein,简称S-100β)和环氧化酶-2(Cyclooxygenase-2,简称COX-2)在1-溴丙烷大鼠亚急性神经损伤中的变化。方法选取健康雄性Wistar大鼠36只,随机分为0ppm(对照组)和1000ppm1-BP染毒组,每组18只,采用口鼻吸入染毒方法,每天染毒6h连续染毒21d。染毒期间隔天称量并记录动物体重,于染毒后第七、十四和二十一天分批次各处死6只动物,采集脑组织和血液(分离血清),每组2只动物分离大脑、垂体、脊髓、坐骨神经和胫腓神经用于病理检测。用ELISA试剂盒检测大鼠脑组织和血清中NSE,S-100β,COX-2等指标的变化。结果与对照组相比,1000ppm染毒21d的染毒组动物体重增长明显减慢(P<0.05),病理检查可见小脑局部浦肯野细胞萎缩、腰髓灰质空泡变性、胫腓神经部分神经纤维肿胀增粗;在染毒的第一周和第三周,大鼠脑组织中NSE、S-100β和COX-2等神经损伤特异性指标与对照组相比显著增高(P<0.05),血清中NSE在第三周有升高趋势,但与对照组相比差异不显著;S-100β在第三周显著增高(P<0.05);而COX-2在前2周显著增高(P<0.05),第三周与对照组差异不显著;脑组织和血清中S-100β的变化显著相关。结论综合实验动物脑组织和血清中神经特异性损伤指标(NSE、S-100β和COX-2)的变化以及病理检查结果,1-BP大鼠染毒模型可以在现有的试验条件和染毒条件下,引起实验动物的亚急性神经损伤。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。
文摘目的探讨神经元特异性烯醇化酶(Neuron Specific Enolase,简称NSE)、星形胶质源性蛋白(S-100βprotein,简称S-100β)和环氧化酶-2(Cyclooxygenase-2,简称COX-2)在1-溴丙烷大鼠亚急性神经损伤中的变化。方法选取健康雄性Wistar大鼠36只,随机分为0ppm(对照组)和1000ppm1-BP染毒组,每组18只,采用口鼻吸入染毒方法,每天染毒6h连续染毒21d。染毒期间隔天称量并记录动物体重,于染毒后第七、十四和二十一天分批次各处死6只动物,采集脑组织和血液(分离血清),每组2只动物分离大脑、垂体、脊髓、坐骨神经和胫腓神经用于病理检测。用ELISA试剂盒检测大鼠脑组织和血清中NSE,S-100β,COX-2等指标的变化。结果与对照组相比,1000ppm染毒21d的染毒组动物体重增长明显减慢(P<0.05),病理检查可见小脑局部浦肯野细胞萎缩、腰髓灰质空泡变性、胫腓神经部分神经纤维肿胀增粗;在染毒的第一周和第三周,大鼠脑组织中NSE、S-100β和COX-2等神经损伤特异性指标与对照组相比显著增高(P<0.05),血清中NSE在第三周有升高趋势,但与对照组相比差异不显著;S-100β在第三周显著增高(P<0.05);而COX-2在前2周显著增高(P<0.05),第三周与对照组差异不显著;脑组织和血清中S-100β的变化显著相关。结论综合实验动物脑组织和血清中神经特异性损伤指标(NSE、S-100β和COX-2)的变化以及病理检查结果,1-BP大鼠染毒模型可以在现有的试验条件和染毒条件下,引起实验动物的亚急性神经损伤。
