DEAE-Sepharose CL-6B离子交换色谱分离α_s酪蛋白的研究

Study on the separation of α_s casein by DEAE-sepharose CL-6B Ion-exchange chromatography

本文建立了酪蛋白中主要组分αs酪蛋白离子交换色谱分离的方法.结果表明,用Z系列φ1.0×40cm色谱柱,DEAE-Sepharose CL-6B为离子交换介质,以含3mol/L尿素,0.1%β-巯基乙醇,pH值为8.0的50mmol/L的tris-HCl缓冲液为平衡液,用含不同浓度NaCl的平衡液进行洗脱,流速为2mL/min,在280nm紫外检测波长下检测,当NaCl浓度为0.3mol/L时洗脱得到αs-酪蛋白,其纯度可达100%.此方法可高效快速的分离出αs酪蛋白,为酪蛋白组分进一步分离及αs酪蛋白相关的功能性食品的开发奠定了基础.

A method of separating αs-casein,the main component of casein,using ion-exchange chromatography was established.The separation was carried out using Z series column（φ1.0×40 cm） filled with DEAE-Sepharose CL-6B ion-exchange media.50 mmol/L of tris-HCl buffer solution（pH 8.0） including 3 mol/L urea and 0.1% β-mercaptoethanol was used as the equilibrium solution and different concentrations of NaCl solution was used as elution solution.The detection wavelength is 280 nm at the rate of 2 mL/min.The results show that purified αs-casein（purity up to 100%） can be obtained when the concentration of NaCl is 0.3 mol/L.The αs-casein fraction with high performance and volant was get using this method.And it builds a foundation for the further separation of casein and the development of functional foods.