Factors determining more efficient large-scale release of a periplasmic enzyme from E-coli using lysozyme
Factors determining more efficient large-scale release of a periplasmic enzyme from E-coli using lysozyme
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DOI：
10.1016/s0168-1656(97)00116-8
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发表时间：
1997-10-02
影响因子：
4.1
通讯作者：
Dunnill, P
中科院分区：
文献类型：
--
作者：
Pierce, JJ;Turner, C;Dunnill, P
大规模使用溶菌酶进行周质释放受到纯酶的成本及其随后在后续下游加工步骤中作为污染物存在的阻碍。在本文中，我们讨论了使用溶菌酶的中试规模回收的周质酶从大肠杆菌。杆菌研究了蔗糖浓度、溶菌酶浓度和菌体浓度对周质酶释放的影响。溶菌酶浓度可以从先前的报道中降低5倍，并且蔗糖浓度从20%（w/v）降低至15%（w/v）允许通过降低粘度来改善离心收获。使用该技术仍然实现了高水平的释放，并且通过优化释放混合物的其他组分获得了产率的进一步提高。结果表明，在不可能使用溶菌酶的情况下，仍然实现了一些释放。结果还表明，相当大比例（高达70%）的溶菌酶在其作用后仍与细胞碎片结合，并与该材料一起被去除。(C)1997年Elsevier Science B.V.
Large scale use of lysozyme for periplasmic release has been impeded by the cost of the pure enzyme and its subsequent presence as a contaminant in later downstream processing steps. In this paper, we discuss the use of lysozyme for pilot scale recovery of a periplasmic enzyme from E. coli. The effects of concentration of sucrose, lysozyme and cells on periplasmic enzyme release were examined. Lysozyme concentration can be reduced 5-fold from previous reports and a reduction in sucrose concentration from 20 to 15% (w/v) allows an improvement in centrifugal harvesting by reducing viscosity. High levels of release were still achieved using this technique and further improvements in yield were obtained by optimising other components of the releasing mixture. Results show that some release is still achieved in circumstances where no lysozyme use is possible. Results also indicate that a substantial proportion (up to 70%) of lysozyme remains bound to the cellular debris after its action and is removed with this material. (C) 1997 Elsevier Science B.V.