Ling Y, Wong H H, Thomas C J, Williams D R, Middelberg A P
Department of Chemical Engineering, University of Adelaide, Australia.
Bioseparation. 1997;7(1):9-15. doi: 10.1023/a:1007900416356.
A new method of poly-beta-hydroxybutyrate (PHB) extraction from recombinant E. coli is proposed, using homogenization and centrifugation coupled with sodium hypochlorite treatment. The size of PHB granules and cell debris in homogenates was characterised as a function of the number of homogenization passes. Simulation was used to develop the PHB and cell debris fractionation system, enabling numerical examination of the effects of repeated homogenization and centrifuge-feedrate variation. The simulation provided a good prediction of experimental performance. Sodium hypochlorite treatment was necessary to optimise PHB fractionation. A PHB recovery of 80% at a purity of 96.5% was obtained with the final optimised process. Protein and DNA contained in the resultant product were negligible. The developed process holds promise for significantly reducing the recovery cost associated with PHB manufacture.
提出了一种从重组大肠杆菌中提取聚-β-羟基丁酸酯(PHB)的新方法,该方法采用均质化和离心,并结合次氯酸钠处理。对匀浆中PHB颗粒和细胞碎片的大小进行了表征,作为均质化次数的函数。利用模拟开发了PHB和细胞碎片分离系统,从而能够对重复均质化和离心进料速率变化的影响进行数值研究。该模拟对实验性能提供了良好的预测。次氯酸钠处理对于优化PHB分离是必要的。最终优化工艺获得了纯度为96.5%时80%的PHB回收率。所得产品中所含的蛋白质和DNA可以忽略不计。所开发的工艺有望显著降低与PHB生产相关的回收成本。