A forward-design approach to increase the production of poly-3-hydroxybutyrate in genetically engineered Escherichia coli.

A forward-design approach to increase the production of poly-3-hydroxybutyrate in genetically engineered Escherichia coli.
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DOI:
10.1371/journal.pone.0117202
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发表时间:
2015
期刊:
影响因子:
3.7
通讯作者:
Freemont P
Freemont P
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Kelwick R;Kopniczky M;Bower I;Chi W;Chin MH;Fan S;Pilcher J;Strutt J;Webb AJ;Jensen K;Stan GB;Kitney R;Freemont P

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生物聚合物,例如聚 3-羟基丁酸酯 (P(3HB)),是在一系列生物体中作为碳储存而产生的,具有与石油衍生塑料相似的特性,但还具有生物降解性和生物相容性的额外优势。尽管有这些优点,但目前 P(3HB) 的生产比石油衍生塑料的生产更昂贵,因此需要更高效的 P(3HB) 生产工艺。在这项研究中,我们描述了几个工程化 P(3HB) 产生操纵子的模型引导设计和实验验证。特别是,我们描述了由双启动子(天然和 J23104)和 RBS(天然和 B0034)设计组成的混合 phaCAB 操纵子的表征。与用真养产碱杆菌 H16 的天然 phaCAB 操纵子改造的大肠杆菌相比,杂交 phaCAB 改造的大肠杆菌在 24 小时时的 P(3HB) 产量高出约六倍。此外,我们描述了利用不可回收废物作为生产 P(3HB) 的低成本碳源。
Biopolymers, such as poly-3-hydroxybutyrate (P(3HB)) are produced as a carbon store in an array of organisms and exhibit characteristics which are similar to oil-derived plastics, yet have the added advantages of biodegradability and biocompatibility. Despite these advantages, P(3HB) production is currently more expensive than the production of oil-derived plastics, and therefore, more efficient P(3HB) production processes would be desirable. In this study, we describe the model-guided design and experimental validation of several engineered P(3HB) producing operons. In particular, we describe the characterization of a hybrid phaCAB operon that consists of a dual promoter (native and J23104) and RBS (native and B0034) design. P(3HB) production at 24 h was around six-fold higher in hybrid phaCAB engineered Escherichia coli in comparison to E. coli engineered with the native phaCAB operon from Ralstonia eutropha H16. Additionally, we describe the utilization of non-recyclable waste as a low-cost carbon source for the production of P(3HB).
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