目的 针对发酵液中L-苹果酸分离纯化工艺存在的回收率低、纯度不足等问题,构建基于离子交换法的分离纯化工艺,以提升产品回收率与纯度。方法 通过静态吸附与洗脱试验筛选D380型离子交换树脂,采用动力学模型和Yoon-Nelson模型分析树脂吸附机制,并结合柱层析动态吸附与洗脱技术,优化上样浓度、流速、层析柱径高比及洗脱剂浓度、流速等关键参数。结果 最佳离子交换柱层析条件：上样浓度为40 g/L、上样体积为60 mL、径高比为1∶5、流速为2 BV/h;洗脱条件为0.4 mol/L氨水、流速1 BV/h。工艺放大6.3倍后,L-苹果酸与富马酸、琥珀酸的分离度显著提升,收率为93.24%;结合后续除盐、结晶步骤,最终产品质量纯度为97.45%,色谱纯度大于99.70%,总工艺收率为79.40%。结论 本研究构建的离子交换法纯化工艺,实现了L-苹果酸的高效分离,具有良好的工业化应用前景。
Abstract
The work aims to develop an ion exchange-based purification process to improve product recovery and purity, so as to address the problems of low recovery and insufficient purity in the separation and purification of L-malic acid from fermentation broth. D380 ion exchange resin was selected through static adsorption and elution experiments, and the adsorption mechanism was analyzed with kinetic models and the Yoon-Nelson model. Key parameters including sample concentration, flow rate, column height-to-diameter ratio, eluent concentration, and flow rate were optimized by dynamic column chromatography. The optimal adsorption conditions were: sample concentration 40 g/L, sample volume 60 mL, height-to-diameter ratio 1∶5, and flow rate 2 BV/h. The optimal elution conditions were 0.4 mol/L ammonia water at a flow rate of 1 BV/h. After 6.3-fold scale-up, the separation resolution between L-malic acid, fumaric acid, and succinic acid was significantly improved, achieving a yield of 93.24%. Combined with subsequent desalination and crystallization steps, the final product exhibited a mass purity of 97.45% and chromatographic purity greater than 99.70%, with an overall process yield of 79.40%. The ion exchange purification process established in this study enables efficient separation of L-malic acid and holds promising potential for industrial application.
关键词
L-苹果酸 /
离子交换法 /
分离纯化 /
工艺放大
Key words
L-malic acid /
ion exchange method /
separation and purification /
process amplification
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基金
天津市合成生物技术创新能力提升行动项目（TSBICIP-PTJJ-006）; 所长领衔科学创新基金 （SZJJ2024-009）
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