Delattre C, Michaud P, Vijayalakshmi M A
Centre for BioSeparation Technology, Vellore Institute of Technology, University, Vellore 632014, Tamil Nadu, India.
J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Jan 15;861(2):203-8. doi: 10.1016/j.jchromb.2007.08.036. Epub 2007 Aug 31.
Fast production and purification of alpha-(1,4)-oligogalacturonides was investigated using a new enzymatic reactor composed of a monolithic matrix. Pectin lyase from Aspergillus japonicus (Sigma) was immobilized on CIM-disk epoxy monolith. Studies were performed on free pectin lyase and immobilized pectin lyase to compare the optimum temperature, optimum pH, and thermal stability. It was determined that optimum temperature for free pectin lyase and immobilized pectin lyase on monolithic support is 30 degrees C, and optimum pH is 5. Monolithic CIM-disk chromatography is one of the fastest liquid chromatographic method used for separation and purification of biomolecules due to high mass transfer rate. In this context, online one step production and purification of oligogalacturonides was investigated associating CIM-disk pectin lyase and CIM-disk DEAE. This efficient enzymatic bioreactor production of uronic oligosaccharides from polygalacturonic acid (PGA) constitutes an original fast process to generate bioactive oligouronides.
利用一种由整体基质组成的新型酶反应器，对α-(1,4)-低聚半乳糖醛酸苷的快速生产和纯化进行了研究。将来自日本曲霉（Sigma）的果胶裂解酶固定在CIM盘环氧整体柱上。对游离果胶裂解酶和固定化果胶裂解酶进行了研究，以比较最佳温度、最佳pH值和热稳定性。结果表明，游离果胶裂解酶和固定在整体柱载体上的果胶裂解酶的最佳温度为30℃，最佳pH值为5。整体CIM盘色谱法由于传质速率高，是用于生物分子分离和纯化的最快的液相色谱方法之一。在此背景下，结合CIM盘果胶裂解酶和CIM盘DEAE，对低聚半乳糖醛酸苷的在线一步生产和纯化进行了研究。这种从聚半乳糖醛酸（PGA）高效酶促生物反应器生产糖醛酸寡糖构成了一种生成生物活性寡糖醛酸苷的原始快速方法。
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