Recombinant protein production in an Escherichia coli reduced genome strain
Recombinant protein production in an Escherichia coli reduced genome strain
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DOI：
10.1016/j.ymben.2006.10.002
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发表时间：
2007-03-01
影响因子：
8.4
通讯作者：
Harcum, Sarah W.
中科院分区：
文献类型：
--
作者：
Sharma, Shamik S.;Blattner, Frederick R.;Harcum, Sarah W.
最近，人们努力通过“基因组手术”来改善大肠杆菌作为重组宿主的特性-删除大肠杆菌K12 MG 1655基因组的大片段而不留下疤痕。这些切除的片段包括K岛，其中含有高比例的转座子，插入序列，隐蔽噬菌体，受损和未知功能的基因。由此产生的多重缺失菌株，命名为大肠杆菌MDS 40，具有比亲本菌株大肠杆菌MG 1655小14%（约700个基因）的基因组。将多重缺失和亲本大肠杆菌菌株在补料分批发酵罐中在基本培养基上培养至高细胞密度，以模拟用于评价生长和重组蛋白生产特征的工业条件。在不同的受控生长速率下定量重组蛋白生产和副产物水平。这些结果表明，多重缺失菌株的生长行为和重组蛋白生产率与亲本菌株密切匹配。因此，多重缺失菌株大肠杆菌MDS 40为进一步基因组减少提供了合适的基础。(c)2006爱思唯尔公司All rights reserved.
Recently, efforts have been made to improve the properties of Escherichia coli as a recombinant host by 'genomic surgery'-deleting large segments of the E coli K12 MG1655 genome without scars. These excised segments included K-islands, which contain a high proportion of transposons, insertion sequences, cryptic phage, damaged, and unknown-function genes. The resulting multiple-deletion strain, designated E coli MDS40, has a 14% (about 700 genes) smaller genome than the parent strain, E coli MG1655. The multiple-deletion and parent E coli strains were cultured in fed-batch fermenters to high cell densities on minimal medium to simulate industrial conditions for evaluating growth and recombinant protein production characteristics. Recombinant protein production and by-product levels were quantified at different controlled growth rates. These results indicate that the multiple-deletion strain's growth behavior and recombinant protein productivity closely matched the parent stain. Thus, the multiple-deletion strain E coli MDS40 provides a suitable foundation for further genomic reduction. (c) 2006 Elsevier Inc. All rights reserved.