Towards improved butanol production through targeted genetic modification of Clostridium pasteurianum.
Towards improved butanol production through targeted genetic modification of Clostridium pasteurianum.
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DOI：
10.1016/j.ymben.2017.01.009
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发表时间：
2017-03
影响因子：
8.4
通讯作者：
Minton NP
中科院分区：
文献类型：
--
作者：
Schwarz KM;Grosse-Honebrink A;Derecka K;Rotta C;Zhang Y;Minton NP
化石燃料储量的减少，加上对其持续开采和开采的环境关切，导致人们作出艰苦努力，以确定获得能源和燃料的可再生途径。一个有吸引力的选择是使用巴氏梭菌将甘油（生物柴油工业的副产品）转化为正丁醇（工业上重要的化学品和潜在的液体运输燃料）。在某些生长条件下，当在甘油上生长时，该梭菌属物种已显示出主要产生正丁醇以及乙醇和1，3-丙二醇。进一步提高C.通过合理的代谢工程改造菌株，因此，在本报告中，我们已经开发并举例说明了一个强大的工具包的代谢工程的C。pasteurianum中，并使用该系统制备了涉及溶剂生产的关键基因即hydA（氢化酶）、雷克斯（氧化还原反应调节剂）和dhaBCE（甘油脱氢酶）的首次报道的框内缺失突变体。这是第一次在C。pasteurianum，能够消除1，3-丙二醇的合成，并证明其生产是必不可少的生长上的甘油作为碳源。雷克斯和hydA的灭活导致正丁醇滴度增加，这代表了提高C. pasteurianum作为这种重要化学品的工业生产的底盘。巴氏梭菌的综合分子工具集。通过dhaBCE KO消除副产物（1，3-丙二醇）的形成。首次报道了梭菌属中主要氢化酶HydA的缺失。雷克斯和hydA的灭活导致正丁醇滴度增加。
Declining fossil fuel reserves, coupled with environmental concerns over their continued extraction and exploitation have led to strenuous efforts to identify renewable routes to energy and fuels. One attractive option is to convert glycerol, a by-product of the biodiesel industry, into n-butanol, an industrially important chemical and potential liquid transportation fuel, using Clostridium pasteurianum. Under certain growth conditions this Clostridium species has been shown to predominantly produce n-butanol, together with ethanol and 1,3-propanediol, when grown on glycerol. Further increases in the yields of n-butanol produced by C. pasteurianum could be accomplished through rational metabolic engineering of the strain. Accordingly, in the current report we have developed and exemplified a robust tool kit for the metabolic engineering of C. pasteurianum and used the system to make the first reported in-frame deletion mutants of pivotal genes involved in solvent production, namely hydA (hydrogenase), rex (Redox response regulator) and dhaBCE (glycerol dehydratase). We were, for the first time in C. pasteurianum, able to eliminate 1,3-propanediol synthesis and demonstrate its production was essential for growth on glycerol as a carbon source. Inactivation of both rex and hydA resulted in increased n-butanol titres, representing the first steps towards improving the utilisation of C. pasteurianum as a chassis for the industrial production of this important chemical. A comprehensive molecular tool set for Clostridium pasteurianum. Abolishment of by-product (1,3-propanediol) formation by dhaBCE KO. First reported deletion of main hydrogenase HydA in Clostridium spp. Inactivation of rex and hydA leads to increased n-butanol titres.