Engineering improved ethanol production in Escherichia coli with a genome-wide approach.
Engineering improved ethanol production in Escherichia coli with a genome-wide approach.
复制标题
工程利用全基因组方法提高了大肠杆菌的乙醇产量。
DOI：
10.1016/j.ymben.2013.01.006
复制
发表时间：
2013
影响因子：
8.4
通讯作者：
R. Gill
中科院分区：
文献类型：
--
作者：
Lauren B. A. Woodruff;Nanette R. Boyle;R. Gill
商品化学品和燃料商业化生产的一个关键挑战是此类分子对微生物宿主的毒性。虽然许多研究试图提高对此类化合物的耐受性，但这些研究大多数是在野生型菌株和与生产条件有很大不同的培养条件下进行的。在这里，我们应用文库富集的多标量分析（SCALE）方法，并在乙醇生产系统中进行生长选择，以并行地将基因组中的所有基因定量映射到乙醇耐受性和生产。为了在乙醇生产系统中进行选择，我们使用先前工程化的大肠杆菌乙醇生产菌株（LW06；ATCC BAA-2466）（Woodruff等人，待出版）作为多标量基因组文库分析的宿主菌株（每个文库具有1、2或4kb重叠基因组片段> 106个克隆）。通过测试单独选择的克隆，我们证实生长选择丰富了具有改善的乙醇耐受性和生产表型的克隆。我们对确定的主要基因（uspC、otsA、otsB）进行了组合测试，以研究它们提高乙醇耐受性或乙醇产量的能力。我们确定 otsA 的过度表达是提高耐受性和生产力表型所必需的，表现最好的菌株相对于亲本生产菌株显示出高达 75% 的改进。
A key challenge to the commercial production of commodity chemical and fuels is the toxicity of such molecules to the microbial host. While a number of studies have attempted to engineer improved tolerance for such compounds, the majority of these studies have been performed in wild-type strains and culturing conditions that differ considerably from production conditions. Here we applied the multiscalar analysis of library enrichments (SCALEs) method and performed a growth selection in an ethanol production system to quantitatively map in parallel all genes in the genome onto ethanol tolerance and production. In order to perform the selection in an ethanol-producing system, we used a previously engineered Escherichia coli ethanol production strain (LW06; ATCC BAA-2466) (Woodruff et al., in press), as the host strain for the multiscalar genomic library analysis (>106clones for each library of 1, 2, or 4kb overlapping genomic fragments). By testing individually selected clones, we confirmed that growth selections enriched for clones with both improved ethanol tolerance and production phenotypes. We performed combinatorial testing of the top genes identified (uspC, otsA, otsB) to investigate their ability to confer improved ethanol tolerance or ethanol production. We determined that overexpression of otsA was required for improved tolerance and productivity phenotypes, with the best performing strains showing up to 75% improvement relative to the parent production strain.
DOI：
10.1073/pnas.92.14.6479
发表时间：
1995-07
影响因子：
11.1
作者：
V. Smith;D. Botstein;Patrick O. Brown
通讯作者：
V. Smith;D. Botstein;Patrick O. Brown
影响因子：
2.7
作者：
Crowe, JH;Crowe, LM;Tablin, F
通讯作者：
Tablin, F
影响因子：
5.6
作者：
Heermann, Ralf;Weber, Arnim;Jung, Kirsten
通讯作者：
Jung, Kirsten