Direct lysozyme separation from egg white by dye membrane affinity chromatography

An affinity membrane from hydrophylised polyethylene hollow fibre as the support matrix was prepared. Red HE‐3B was immobilised on the membrane and the adsorption behaviour of pure lysozyme solutions and homogenised egg white was investigated. Dye density (1.7 μmol ml⁻¹) and maximum binding capacity (26 mg lysozyme ml⁻¹) are comparable to those of commercial gel matrices. Dynamic binding capacity did not change when residence time was reduced from 3 to 1 min. A method for direct lysozyme separation from egg white was developed, with a productivity of 12 kg lysozyme m⁻³ h⁻¹. The purity of the eluted lysozyme, as determined by HPLC, was 88%, with a recovery of 92%. Dynamic capacity was kept constant at 70% of the maximum binding capacity for at least 10 cycles through membrane regeneration with 0.1 M NaOH and 1 M NaCl. Functional properties of egg white, as judged by viscosity and foaming capacity measurements, did not change after the chromatographic lysozyme depletion. © 1999 Society of Chemical Industry     

层析法分离纯化纳豆激酶的技术参数研究

采用柱层析技术对盐析后的纳豆激酶发酵液进行分离纯化,以确定层析分离纳豆激酶的技术参数.对比了Sephadex G-50及Sephadex G-7凝胶层析分离纳豆激酶的效果,并进行了不同pH值条件下的层析效果分析.结果表明,在pH值为6.4时,经Sephadex G-75凝胶层析及SP Sepharose FF离子交换层析后,能够得到较高纯度的纳豆激酶,使纳豆激酶纯化了9.2倍,回收率为51.5％.SDS-PAGE电泳显示所得纳豆激酶为单一蛋白带,其分子质量为28ku.     

Fractionation of fungal pectic enzymes by immobilised metal ion affinity chromatography

A juice-clarifying fraction that will not produce methanol was obtained from a commercial pectic enzyme by separating the pectin lyase (EC4.2.2.10) from pectinesterase (EC3.1.1.11). This was achieved by immobilised metal ion affinity chromatography (IMAC) on Sepharose—iminodiacetic acid—Cu(II). Pectin lyase did not bind to the chromatographic matrix at pH 8.0, while pectinesterase was retained and only eluted when the pH of the buffer was brought down to 3.0. Polygalacturonase activity (EC3.2.1.15) was found in both fractions thus suggesting that IMAC can discriminate between two forms of that enzyme Both fractions have the ability to clarify apple juice, the first without producing methanol. The behaviour of each of the above enzymes in IMAC suggests that pectin lyase lacks accessible histidine residues, while pectinesterase could have one or more of them.     

离子交换法分离纯化甘露低聚糖

采用离子交换树脂法分离纯化甘露低聚糖,由实验得到最佳分离条件为进样量为10 mL、柱高为600 mm、流速为2 mL/min、温度为60 ℃,在此条件下甘露低聚糖的纯度为68.4%.     

置换色谱法分离纯化大豆磷脂的研究

采用置换色谱法分离纯化了大豆磷脂,研究表明:以二氯甲烷-甲醇为流动相、置换剂选择10%乙醇胺、流速0.5 mL/min、上样量为110 mg,在此置换条件下,获得的PE、PC、PI含量可进一步富集,分别为从52.9%、8.7%、30%增长至78.8%、45.3%、84%,其回收率分别为48.4%、81.6%、65.9%。     

鸡蛋壳溶菌酶分离纯化工艺研究

以鲜鸡蛋壳为原料,1.0％NaCl(pH3.0,40℃)提取溶菌酶, 724大孔阳离子树脂色谱纯化,10％(NH_4)_2SO_4洗脱,盐析 结晶。用羧甲基纤维素CM52柱色谱分析,显示产品纯度 较高。     

红蘑多糖的纤维柱层析分离纯化

选择野生红蘑作为提取多糖的原料,采用预处理后的DEAE-52纤维素粉末制成OH-型柱对红蘑中的多糖进行分离纯化,结果表明:在上样量为200mg、洗脱速度为0.5mL/min的条件下洗脱,得到4个主峰,依次为GRPⅠ、GRPⅡ、GRPⅢ和GRPⅣ,其中GRPⅠ出现在水洗脱部分,GRPⅡ、GRPⅢ和GRPⅣ分别出现在0.1mol/LNaCl、0.3mol/LNaCl、1.0mol/LNaCl洗脱部分。经计算,GRPⅠ、GRPⅡ、GRPⅢ和GRPⅣ分别占12.6%、8.7%、22.3%和27.6%。     

硅胶柱层析法分离纯化甘露低聚糖

采用硅胶柱层析法分离纯化甘露低聚糖,由实验得到最佳分离条件为:进样量为10mL,柱高为600mm,流速为1mL/min,温度为60℃,在此条件下甘露低聚糖的纯度为71.34%.     

One-step purification of lactoperoxidase from bovine milk by affinity chromatography

Sulphanilamide was determined to be a new inhibitor of lactoperoxidase (LPO) with an IC₅₀ of 0.848.10⁻⁵ M. The K_i for sulphanilamide was determined to be 3.57.10⁻⁵ M and sulphanilamide showed competitive inhibition, which makes it a suitable ligand for constructing a Sepharose 4B-l-tyrosine affinity matrix. The affinity matrix was synthesised by coupling sulphanilamide as the ligand and l-tyrosine as the spacer arm to a cyanogen bromide (CNBr)-activated-Sepharose 4B matrix. Lactoperoxidase was purified 409-fold from the synthesized affinity matrix in a single step, with a yield of 62.3% and a specific activity of 40.9 EU/mg protein. The enzyme activity was measured using ABTS as a chromogenic substrate (pH 6.0). The degree of LPO purification was monitored by SDS–PAGE and its R_z (A₄₁₂/A₂₈₀) value. The R_z value for the purified LPO was found to be 0.7. Maximum binding was achieved and K_m and V_max values were determined.     

应用高速逆流色谱分离纯化芦荟苷

利用高速逆流色谱对芦荟中的蒽醌类活性成分芦荟苷进行了制备型分离研究。溶剂系统:氯仿-正丁醇-甲醇-水(4:0.28:3:2,V/V),上相为固定相,下相为流动相,转速870r/min,流速2mL/min。经TLC和HPLC分析,分离组分芦荟苷A和B的纯度分别达到96%和93%。     

离子交换层析法分离纯化玉米降血压肽的研究

以玉米蛋白为原料酶解得到了具有降血压活性酶解液,为了得到高活性、高纯度的降血压肽,采用离子交换层析法对酶解液进行了分离纯化。主要研究了离子强度、离子强度梯度以及上样量对分离效果的影响,得到最佳条件:NaCl强度为2mol/L;离子强度梯度为0.024mol/L·min;最大上样量为300mg。在此条件下分离得到的组分,其抑制ACE活性达到90%,含量为25g/100g。     

分离纯化新技术亲和层析

本文介绍了亲和层析的一般问题 ,综述了固定化金属亲和层析、免疫亲和层析和其它一些类型的亲和层析的原理和应用     

柱层析分离纯化功能性红曲米中的有效成分

为深入研究功能性红曲米浓缩物中有效成分的性质及其功能,采用硅胶柱层析法(依次用乙酸乙酯:石油醚(1∶1,v∶v)、乙酸乙酯、无水甲醇、蒸馏水进行洗脱)对功能性红曲米浓缩物中的活性成分进行分离纯化,得到红曲黄色素、洛伐他汀粗品、醇溶性红曲红色素、水溶性红曲红色素4种组分;并采用紫外-可见扫描和高效液相色谱法对4种组分进行简单的性质结构表征.     

离子交换层析法分离猪股骨降血压肽的研究

猪股骨酶解液经超滤、凝胶层析初步分离后,为了得到高活性且纯度较高的降血压肽,采用离子交换层析对凝胶层析分离后的高活性组分进一步分离。通过对洗脱液p H、离子强度、流速三个因素进行研究,确定了最佳分离条件:以p H=6.0、0.05mol/L磷酸盐缓冲液为A液、B液为含1mol/LNa Cl的p H=6.0、0.05mol/L磷酸盐缓冲液,洗脱流速为0.8m L/min。结果显示:离子交换层析分离得3个组分,其中组分1的活性最高,其IC50值为0.1012mg/m L;再利用凝胶层析将组分1进行脱盐,其峰2的IC50值达到0.0836mg/m L,脱盐率达到86.60%±0.5%。      

离子交换层析法分离猪股骨降血压肽的研究

猪股骨酶解液经超滤、凝胶层析初步分离后,为了得到高活性且纯度较高的降血压肽,采用离子交换层析对凝胶层析分离后的高活性组分进一步分离。通过对洗脱液p H、离子强度、流速三个因素进行研究,确定了最佳分离条件:以p H=6.0、0.05mol/L磷酸盐缓冲液为A液、B液为含1mol/LNa Cl的p H=6.0、0.05mol/L磷酸盐缓冲液,洗脱流速为0.8m L/min。结果显示:离子交换层析分离得3个组分,其中组分1的活性最高,其IC50值为0.1012mg/m L;再利用凝胶层析将组分1进行脱盐,其峰2的IC50值达到0.0836mg/m L,脱盐率达到86.60%±0.5%。     

膜分离与大孔树脂联用技术纯化茶皂素

采用膜分离与大孔树脂联用技术纯化茶皂素。粗茶皂素经陶瓷膜和360Da纳滤膜初步分离浓缩,得率为62.1%,纯度为79%;根据静态和动态吸附筛选试验,选择大孔树脂AmberliteXAD7HP对茶皂素进一步纯化,通过单因素试验,确定最佳工艺参数为:上样流速0.5 mL/min、上样液浓度30mg/mL;以10%,40%,70%的乙醇溶液进行梯度洗脱,洗脱剂流速1mL/min,洗脱液体积为3BV,该条件下纯化,茶皂素最终得率为55.3%,纯度可达95%。该试验表明膜分离与大孔树脂联用技术可得到高纯度的茶皂素,是一种可工业化推广的方法。     

高速逆流色谱法分离纯化新疆圆柏枝叶中的黄酮类成分

目的:建立高速逆流色谱分离纯化新疆圆柏枝叶中的黄酮类成分的方法。方法:以氯仿∶甲醇∶正丁醇∶水（4∶3∶0.5∶2）为溶剂系统,上相为固定相,下相为流动相,在流速2.0 m L/min、转速850 r/min、检测波长254 nm的条件下,对新疆圆柏中的黄酮类成分进行分离纯化。结果:从新疆圆柏提取物中分离纯化得到3个化合物:异槲皮苷（a,91.89%）、槲皮苷（b,98.45%）、槲皮素-3-O-（6″-O-乙酰基）-β-D-吡喃葡萄糖苷（c,95.35%）,其中c为首次从圆柏属植物中分离得到的化合物。结论:该法快速简便,能够高效分离新疆圆柏中的黄酮类成分。      

高速逆流色谱法分离纯化红景天苷、葛根素和淫羊藿苷3种标准样品

目的采用高速逆流色谱技术分离纯化红景天苷、葛根素和淫羊藿苷3种标准样品。方法采用溶剂体系正丁醇:乙酸乙酯:水(2:3:5,V:V:V)分离红景天粗提物;采用溶剂体系正丁醇:乙酸乙酯:水(1:2:3,V:V:V)分离葛根粗提物;采用氯仿:甲醇:水(4:3.5:2,V:V:V)分离淫羊藿苷粗提物,并采用高效液相色谱法对所得单体进行纯度检测。结果所得红景天苷、葛根素、淫羊藿苷标准样品纯度分别为98.9%、99.8%和99.5%。将核磁共振(nuclear magnetic resonance,NMR)13C-NMR数据归属并与相关文献比对,进一步确认了该3种物质的结构。结论红景天苷、葛根素、淫羊藿苷3种样品满足GB/T 15000.3-2008标准样品工作导则的要求,可用于相关药品检测方法的校正和相关产品的质量控制。     

两种不同离子层析法分离鸡蛋溶菌酶的比较研究

以鸡蛋溶菌酶的主要分离方法—阳离子交换法与阴离子交换法为对象,从分离的回收率、鸡蛋溶菌酶纯度以及样品处理周期等方面进行比较研究。研究结果显示,两种方法均能较好地分离鸡蛋溶菌酶。阴离子交换法分离出的鸡蛋溶菌酶纯度更高,纯度>90%,并且分离周期短,操作更方便,但其回收率为20·7%,而阳离子交换法的为31·1%,阴离子交换法更适合于制备高纯度的蛋清溶菌酶。