Design and characterisation of synthetic operons for biohydrogen technology.
Design and characterisation of synthetic operons for biohydrogen technology.
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DOI：
10.1007/s00203-016-1322-5
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发表时间：
2017-04
影响因子：
2.8
通讯作者：
Sargent F
中科院分区：
文献类型：
--
作者：
Lamont CM;Sargent F
生物氢由许多微生物系统产生，并且常用的宿主细菌大肠杆菌在发酵条件下天然产生氢。一种工程化额外的氢产生途径的方法是将非天然氢化酶引入E.杆菌一个有吸引力的候选者是来自Ralstonia eutropha的可溶性[NiFe]-氢化酶，其已被证明将NADH/NAD+生物化学直接连接到氢代谢，这是E.大肠杆菌不起作用。在这项工作中，设计了三个合成操纵子，编码可溶性氢化酶和两种不同的酶成熟酶系统。有趣的是，利用这个系统，重组可溶性氢化酶被发现是由天然的E。coli [NiFe]-氢化酶装配机器，反之亦然，合成的成熟酶操纵子能够补充E.大肠杆菌氢化酶生物合成缺陷型突变体。异源表达的可溶性氢化酶被发现是活跃的，并显示出在体内产生生物氢。
Biohydrogen is produced by a number of microbial systems and the commonly used host bacterium Escherichia coli naturally produces hydrogen under fermentation conditions. One approach to engineering additional hydrogen production pathways is to introduce non-native hydrogenases into E. coli. An attractive candidate is the soluble [NiFe]-hydrogenase from Ralstonia eutropha, which has been shown to link NADH/NAD+ biochemistry directly to hydrogen metabolism, an activity that E. coli does not perform. In this work, three synthetic operons were designed that code for the soluble hydrogenase and two different enzyme maturase systems. Interestingly, using this system, the recombinant soluble hydrogenase was found to be assembled by the native E. coli [NiFe]-hydrogenase assembly machinery, and, vice versa, the synthetic maturase operons were able to complement E. coli mutants defective in hydrogenase biosynthesis. The heterologously expressed soluble hydrogenase was found to be active and was shown to produce biohydrogen in vivo.