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首页> 外文期刊>Biotechnology Progress >Enhanced Biodegradation of Toxic Organophosphate Compounds Using Recombinant Escherichia coli with Sec Pathway-Driven Periplasmic Secretion of Organophosphorus Hydrolase
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Enhanced Biodegradation of Toxic Organophosphate Compounds Using Recombinant Escherichia coli with Sec Pathway-Driven Periplasmic Secretion of Organophosphorus Hydrolase

机译：重组大肠杆菌与Sec途径驱动的周质分泌的有机磷水解酶增强的有毒有机磷酸酯化合物的生物降解。

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Although Escherichia coli can be genetically engineered to degrade environmental toxic organophosphate compounds (OPs) to nontoxic materials,a critical problem in such whole cell systems is limited substrate diffusion.The present work examined whether periplasmic expression of organophosphorus hydrolase (OPH) resulted in better whole cell enzymatic activity compared to standard cytosolic expression.Recombinant OPH periplasmic expression was achieved using the general secretory (sec) pathway with the pelB signal sequence.We found that while total OPH activity in periplasmic-expressing cell lysates was lower compared to that in cytosolic-expressing cell lysates whole cell OPH activity was 1.8-fold greater at 12 h post-induction in the periplasmic-expressing cells as a result of OPH translocation into the periplasmic space (approx 67% of whole cell OPH activity was found in the periplasmic fraction).These data suggest that E.coli engineered to periplasmically secrete OPH via the sec pathway may provide an improved whole cell biodegradation system for destruction of environmental toxic OPs.
机译：尽管可以通过基因工程改造大肠杆菌以将环境毒性有机磷酸酯化合物（OPs）降解为无毒物质，但在这种全细胞系统中的一个关键问题是底物扩散受限。目前的工作研究了有机磷水解酶（OPH）的周质表达是否导致整体质量更好细胞的酶活性与标准胞质表达相比。使用一般分泌（sec）途径与pelB信号序列实现重组OPH胞质表达。我们发现，表达胞质的细胞裂解液中的总OPH活性低于胞质中的表达细胞裂解物诱导表达后的12 h，表达周质的细胞的全细胞OPH活性高1.8倍，这是因为OPH易位到周质空间（在周质中发现了约67％的全细胞OPH活性）这些数据表明大肠杆菌经工程设计可通过sec途径周质分泌OPH可以提供改进的全细胞生物降解系统，以破坏环境有毒OP。

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