Windhorst Carina, Gescher Johannes
1Department of Applied Biology, Institute for Applied Biosciences, Karlsruhe Institute of Technology, Karlsruhe, Germany.
2Institute for Biological Interfaces, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany.
Biotechnol Biofuels. 2019 Jun 28;12:163. doi: 10.1186/s13068-019-1512-x. eCollection 2019.
is the best-studied knallgas (also termed hydrogen oxidizing) bacterium and provides a model organism for studying the production of the storage polymer polyhydroxybutyrate (PHB). Genetically engineered strains could be applied for the autotrophic production of valuable chemicals. Nevertheless, the efficiency of the catalyzed processes is generally believed to be lower than with acetogenic bacteria. Experimental data on the potential efficiency of autotrophic production with are sparse. Hence, this study aimed at developing a strain for the production of the bulk chemical acetoin from carbon dioxide and to analyze the carbon and electron yield in detail.
We developed a constitutive promoter system based on the natural PHB promoter of this organism. Codon-optimized versions of the acetolactate dehydrogenase () and acetolactate decarboxylase () from were cloned under control of the PHB promoter in order to produce acetoin from pyruvate. The production process's efficiency could be significantly increased by deleting the PHB synthase . Further deletion of the other PHB synthase encoded in the genome () led to a strain that produced acetoin with > 100% carbon efficiency. This increase in efficiency is most probably due to a minor amount of cell lysis. Using a variation in hydrogen and oxygen gas mixtures, we observed that the optimal oxygen concentration for the process was between 15 and 20%.
To the best of our knowledge, this study describes for the first time a highly efficient process for the chemolithoautotrophic production of the platform chemical acetoin.
是研究最为深入的爆鸣气(也称为氢氧化细菌)细菌,为研究储存聚合物聚羟基丁酸酯(PHB)的生产提供了一种模式生物。基因工程菌株可用于自养生产有价值的化学品。然而,一般认为催化过程的效率低于产乙酸细菌。关于利用自养生产潜在效率的实验数据很少。因此,本研究旨在开发一种从二氧化碳生产大宗化学品乙偶姻的菌株,并详细分析碳和电子产量。
我们基于该生物体的天然PHB启动子开发了一种组成型启动子系统。来自的乙酰乳酸脱氢酶()和乙酰乳酸脱羧酶()的密码子优化版本在PHB启动子的控制下进行克隆,以便从丙酮酸生产乙偶姻。通过删除PHB合酶,生产过程的效率可显著提高。进一步删除基因组中编码的另一种PHB合酶(),得到了一种以>100%的碳效率生产乙偶姻的菌株。效率的提高很可能是由于少量细胞裂解。通过改变氢气和氧气的混合气体,我们观察到该过程的最佳氧气浓度在15%至20%之间。
据我们所知,本研究首次描述了一种用于平台化学品乙偶姻化学自养生产的高效过程。