Fusion expression of m - centrin in e . coil bl21 was performed by induction of fptg . fusion protein was digested by ppase and was purified by gst chelating affinity chromatography and ion exchange chromatography . the final products were checked by sds - page gel 融合蛋白gst - m - centrin菌体经过超声波裂解后得到的上清夜经过gst亲和层析后用prescissionprotease ( ppase )酶切，酶切产物再次经过gst亲和层析和hitrapq阴离子交换层析两步柱层析纯化后，得到纯度较高的的m - centrin 。
Then enzyme was purified with a deae - cellulose ( 5 . 5x50cm ) column , a toyopearl hw - 65 ( 5 . 5 x 50cm ) column and a sephadex g - 200 ( 5 . 5 x 80cm ) column . finally , the enzyme was purified for 10 folds with the recovery of 17 . 4 % . page showed a single band for the purified creatinase 3 、肌酸水解酶的提纯酶在硫酸铵饱和度为40 80之间完全沉淀，先后经过deae - cellulose离子层析柱、 toyopearlhw - 65疏水层析柱、 sephadexg - 200分子筛层析柱层析，最终使酶提纯10倍，最终得率为17 . 4 。
Many kinds of chromatograph including silica gel , d101 resin , sephadex l - 20 gel , rp - 8 and hplc are used in extracting and separating chemical constituents form these plants . certainly , they need to select the best eluate by tlc . some technique including ms ( ei - ms , fab - ms ) , id and 2d nmr ( cosy , roesy , hmbc , hmqc ) been used in identifying these chemical structures 通过各种硅胶柱层析，吸附树脂， sephadex系列凝胶树脂以及rp - 8类反向柱等层析材料以及高效液相色谱等技术，利用不同的洗脱体系对这两种植物的化学成分进行分离得到纯的单体化合物，然后利用ms ( ei - ms ， fab - ms ) 、一维nmr 、二维nmr ( cosy ， roesy ， hmbc ， hmqc )等技术对这些化合物进行结构鉴定，利用ir和uv对这些化合物进行了表征。
Both hainantoxin - iv ( hntx - iv ) and . hainantoxin - v ( hntx - v ) w ere peptide toxins isolated from the venom of the spider seleconosmia hainana by means of ion - exchange chromatograph and reverse phase hplc . hntx - iv , 3989 da , contained 35 amino acid residues with three disulfide bonds , while the molecular weight of hntx - v was 3972 da and had three disulfide bonds , too 海南捕鸟蛛毒素- ( hntx - )和海南捕鸟蛛毒素- ( hntx - )是从海南捕鸟蛛粗毒中经阳离子交换和反相hplc柱层析分离出来的两种神经毒素， maldi - tof质谱鉴定前者相对分子质量为3 ， 989da ，含有三对二硫键，由35个氨基酸残基组成；后者相对分子质量为3 ， 972da ，与前者只相差16个质子，也含有三对二硫键。
To isolate and purify dnaase in the earthworm first , the tissue extract of earthworm was prepared by dissolving the earthworm with sucrose and denaturing the protein with low ph buffer . then dnaase was purified by denaturing the protein with higher temperature . the following steps were ammonium sulfate precipitation , deae - cellulose ( de52 ) chromatography and filtration by ultra - filter membrane 双胸蚓组织中dna酶的分离纯化采用蔗糖溶解双胸蚓，并选择性酸变性制备双胸蚓组织粗提取液，再经选择性热变性、硫酸铵分段盐析、 deae ?纤维素( de52 )柱层析及超滤膜分级分离对双胸蚓组织中dna酶进行分离纯化。
6 - phosphogluconate dehydrogenase ( 6 - pgadase , ec 1 . 1 . 1 . 44 ) was isolated by homogenate , ammunium sulfate fractionation , deae - sepharose chromatography , blue - sepharose affinity chromatography and gel filtration with sephadex g - 200 from bacillus subtilis , and some properties of the enzyme had been studied . a 113 . 8 - fold purification was obtained with a 8 . 2 % yield . the purified enzyme moved as a single electrophoretic band in page 将枯草芽孢杆菌超声波破壁后的粗提取物进行分段盐析、 deae - sepharose阴离子交换柱层析， blue - sepharosecl - 6b特异结合柱层析和sephadexg - 200凝胶过滤等纯化步骤，得到聚丙烯酰胺凝胶电泳为单一蛋白区带，比活为1 . 46u mg的酶制剂。
The enzyme retained full activity after being treated at room temperature for 1 hour at ph between 4 . 0 and 11 . 5 . the enzyme can be incubated at 50 for 4h with only less 50 percent loss of activity and is stable in the frozen state . when streptomyces griseus atcc14811 was cultured in 10 . 3 % sucrose yeme liquid medium , production of extracellular cholesterol oxidase increased for 5 days before decrease 利用硫酸铵盐析及deae -纤维素离子交换柱层析提取纯化灰色链霉菌atcc14811发酵上清液中的胆固醇氧化酶，理化性质研究表明酶作用晟适ph为8 . 0 ，最适温度为45 ， ph稳定范围在ph4 . 0 - 11 . 5之间，在50条件下保温4h ，仍保留54酶活力。
According to the polarity , the residue was isolated with petroleum ether , chloroform , ethyl acetate , and n - buoh , respectively . the n - buoh fraction , confirmed as neuroactive component , was subjected to sephadex lh - 20 column chromatography to provide an extract fraction , as a buff powder , which could induce neurite outgrowth in rat pheochromocytoma pc12 cells in a dose - dependent manner up to 50 mg / l 将菟丝子干粉用75乙醇浸泡后，减压蒸干后得到褐色浆状物，经石油醚、氯仿、乙酸乙酯、正丁醇萃取，经柱层析后，再用葡聚糖凝胶对有效成份进一步纯化，获得了菟丝子中能诱导pc12细胞分化的活性组分。
The bioactive strain ' s fermentation product was isolated and purified primarily using methods of solvent extraction , acid - alkali extraction , ab - 8 macropore absorption chromatography , 1 x 007 positive ion exchange chromatography , the result showed the purification product has prominent bioactivity inhibit staphalococcus aureus 中国热带农业科学院、华南热带农业大学2003届硕士研究生采用有机剂抽提法、酸碱抽提法、大孔吸附树脂柱层析和阳离子交换树脂柱层析等方法对hsl 306菌株发酵产物进行了初步分离纯化。
The crude extraction of dry pea seed , which was obtained through marination , homogenization , filtration , centrifugation and other methods , was precipitated by 50 mmol / l ( final concentration ) mgq2 . the pellet was chromatographed on aca ^ gel filtration and deae - cellulose 52 ( de 52 ) anion - exchanger . single eluting peak containing ferritin was obtained finally 2 、豌豆种子铁蛋白的纯化豌豆种子经浸泡、匀浆、过滤、离心等操作得到豌豆铁蛋白粗提液，再用终浓度为50mmol / l的mgcl _ 2盐析等处理后，经aca _ ( 22 )凝胶过滤柱层析和deae -纤维素52 ( de52 )阴离子交换柱层析等方法进行纯化，得到单一的豌豆铁蛋白洗脱峰。
