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采用大孔树脂色谱法从大肠杆菌发酵液中分离和纯化 L-蛋氨酸。

Separation and purification of l-methionine from E. coli fermentation broth by macroporous resin chromatography.

机构信息

Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China; Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou 310014, China.

Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China; Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou 310014, China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Mar 15;1110-1111:108-115. doi: 10.1016/j.jchromb.2019.02.016. Epub 2019 Feb 15.

Abstract

Methionine is an essential sulfur-containing amino acid for organisms. The separation and purification are important for the production of l-methionine from fermentation broth. In this work, the adsorption properties for l-methionine separation of ten macroporous resins were firstly evaluated. Macroporous cation resin D72 showed the best adsorption capacity (52.37 mg/g) and desorption rate (99.12%). The adsorption kinetics of l-methionine on D72 resin followed the pseudo second-order model and adsorption isotherm fitted well to the Sips model, the adsorption process mainly followed a physisorption mechanism. D72 resin packed columns were then used in the batch separation of fermentation broth, the operation parameters were optimized during the dynamic adsorption and desorption experiments. Under the optimized conditions: fermentation broth adjusted to pH = 2, loading flow rate at 2 BV/h, loading volume 55 mL, 1 mol/L NH·HO as eluent, elution flow rate at 2 BV/h, column height-diameter ratio at 14:1, excellent recovery and purity of l-methionine (82.37% and 85.69%, respectively) could be achieved.

摘要

蛋氨酸是生物体必需的含硫氨基酸。分离和纯化对于从发酵液中生产 L-蛋氨酸非常重要。在这项工作中,首先评估了十种大孔树脂对 L-蛋氨酸分离的吸附性能。大孔阳离子树脂 D72 表现出最好的吸附容量(52.37mg/g)和洗脱率(99.12%)。L-蛋氨酸在 D72 树脂上的吸附动力学符合拟二级动力学模型,吸附等温线很好地符合 Sips 模型,吸附过程主要遵循物理吸附机制。然后使用 D72 树脂填充柱在分批分离发酵液,在动态吸附和解吸实验中优化了操作参数。在优化条件下:发酵液 pH 值调至 2,上样流速为 2BV/h,上样体积 55mL,1mol/L NH·HO 作为洗脱液,洗脱流速为 2BV/h,柱高径比为 14:1,可以获得优异的 L-蛋氨酸回收率和纯度(分别为 82.37%和 85.69%)。

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