Development of genetically stable Escherichia coli strains for poly(3-hydroxypropionate) production.

Development of genetically stable Escherichia coli strains for poly(3-hydroxypropionate) production.
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用于聚(3-羟基丙酸酯)生产的遗传稳定大肠杆菌菌株的开发

DOI:
10.1371/journal.pone.0097845
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发表时间:
2014
期刊:
影响因子:
3.7
通讯作者:
Zhao G
Zhao G
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Gao Y;Liu C;Ding Y;Sun C;Zhang R;Xian M;Zhao G

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聚(3-羟基丙酸酯)(P3HP)是一种可生物降解、生物相容的热塑性塑料。在我们之前的研究中,我们构建了一条在重组大肠杆菌中生产P3HP的途径。将P3HP合成途径中的7个外源基因分别携带在两个载体上。然而,P3HP的产生受到了由于质粒丢失而导致的菌株不稳定性的严重抑制。本论文采用了染色体基因整合和基于氨基酸合成的质粒成瘾系统(PAS)两种策略,构建了一株遗传稳定的菌株。最后,这两种方法的组合得到了最好的结果。该菌株在染色体上携带部分P3HP合成基因,在质粒上携带部分P3HP合成基因,并携带酪氨酸营养缺陷型PAS。在好氧补料分批发酵中,该菌产甘油25.7g/L P3HP,比双质粒发酵提高2.5倍左右。据我们所知,这是利用廉价碳源生产的最高P3HP产品。
Poly(3-hydroxypropionate) (P3HP) is a biodegradable and biocompatible thermoplastic. In our previous study, a pathway for P3HP production was constructed in recombinant Esecherichia coli. Seven exogenous genes in P3HP synthesis pathway were carried by two plasmid vectors. However, the P3HP production was severely suppressed by strain instability due to plasmid loss. In this paper, two strategies, chromosomal gene integration and plasmid addiction system (PAS) based on amino acid anabolism, were applied to construct a genetically stable strain. Finally, a combination of those two methods resulted in the best results. The resultant strain carried a portion of P3HP synthesis genes on chromosome and the others on plasmid, and also brought a tyrosine-auxotrophy based PAS. In aerobic fed-batch fermentation, this strain produced 25.7 g/L P3HP from glycerol, about 2.5-time higher than the previous strain with two plasmids. To the best of our knowledge, this is the highest P3HP production from inexpensive carbon sources.
DOI: 10.1128/jb.153.3.1424-1431.1983
发表时间: 1983-01-01
影响因子: 3.2
作者:
GAY, P;LECOQ, D;HOCH, JA
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