Calado C R, Hamilton G E, Cabral J M, Fonseca L P, Lyddiatt A
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Lisboa, Portugal.
Bioseparation. 2001;10(1-3):87-97. doi: 10.1023/a:1012464218516.
The recovery of cutinase of Fusarium solani pisi produced by the yeast Saccharomyces cerevisiae was studied in a fluidised bed adsorption system directly integrated with a productive fermenter (so-called direct product sequestration; DPS). The relative efficiency of this system was compared with the one of a conventional purification process by discrete sequences of fermentation, broth clarification, ultrafiltration and fixed bed anion exchange chromatography. By direct product sequestration of the extracellular heterologous cutinase it was possible, through only one unit operation: (i) to perform broth clarification, (ii) to obtain a high cutinase concentration factor, and (iii) to recover cutinase with a specific activity that equalled that obtained with the conventional purification process. It was also possible (iv) to substantially reduce the total process time, (v) to improve the overall yield, and (vi) to increase cutinase productivity. Furthermore, the procedure outlined is suitable for large scale bioprocess exploitation.
在与生产发酵罐直接集成的流化床吸附系统（即所谓的直接产物截留；DPS）中，研究了酿酒酵母生产的豌豆镰刀菌角质酶的回收情况。将该系统的相对效率与通过发酵、肉汤澄清、超滤和固定床阴离子交换色谱等离散步骤组成的传统纯化工艺的效率进行了比较。通过直接产物截留细胞外异源角质酶，仅通过一个单元操作就可以：（i）进行肉汤澄清，（ii）获得高角质酶浓缩系数，（iii）回收比活性与传统纯化工艺相当的角质酶。还可以（iv）大幅缩短总工艺时间，（v）提高总产率，以及（vi）提高角质酶生产率。此外，所述程序适用于大规模生物工艺开发。
