3-Methyl-1-butanol production in Escherichia coli: random mutagenesis and two-phase fermentation.

3-Methyl-1-butanol production in Escherichia coli: random mutagenesis and two-phase fermentation.
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DOI:
10.1007/s00253-009-2401-1
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发表时间:
2010-04
影响因子:
5
通讯作者:
Liao, James C.
Liao, James C.
中科院分区:
工程技术2区
文献类型:
--
作者:
Connor, Michael R.;Cann, Anthony F.;Liao, James C.

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出于对能源和环境的考虑,人们对利用可再生资源生产生物燃料的兴趣日益浓厚。近年来,从2-酮酸途径生产高链醇的研究取得了重大进展。在本文中,我们展示了一种诱变方法,通过利用l-亮氨酸生物合成的选择压力和筛选增加的酒精产量,开发了一种生产3-甲基-1-丁醇的大肠杆菌菌株。用与L -亮氨酸结构类似的4-aza-d - L -亮氨酸进行随机诱变和选择,获得了一株能产生4.4 g/L 3-甲基-1-丁醇的新菌株。通过对宿主对3-甲基-1-丁醇敏感性的研究,确定了两阶段发酵工艺,使3-甲基-1-丁醇滴度达到9.5 g/L, 60 h后葡萄糖产量为0.11 g/g。
Interest in producing biofuels from renewable sources has escalated due to energy and environmental concerns. Recently, the production of higher chain alcohols from 2-keto acid pathways has shown significant progress. In this paper, we demonstrate a mutagenesis approach in developing a strain of Escherichia coli for the production of 3-methyl-1-butanol by leveraging selective pressure toward l-leucine biosynthesis and screening for increased alcohol production. Random mutagenesis and selection with 4-aza-d,l-leucine, a structural analogue to l-leucine, resulted in the development of a new strain of E. coli able to produce 4.4 g/L of 3-methyl-1-butanol. Investigation of the host’s sensitivity to 3-methyl-1-butanol directed development of a two-phase fermentation process in which titers reached 9.5 g/L of 3-methyl-1-butanol with a yield of 0.11 g/g glucose after 60 h.
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