Increasing the metabolic capacity of Escherichia coli for hydrogen production through heterologous expression of the Ralstonia eutropha SH operon.
Increasing the metabolic capacity of Escherichia coli for hydrogen production through heterologous expression of the Ralstonia eutropha SH operon.
复制标题
DOI：
10.1186/1754-6834-6-122
复制
发表时间：
2013-08-26
影响因子：
6.3
通讯作者：
Hallenbeck PC
中科院分区：
文献类型：
--
作者：
Ghosh D;Bisaillon A;Hallenbeck PC
发酵制氢是一种有吸引力的可持续生产这种未来能源载体的手段，但受到低产率的阻碍。一种可能的解决方案是使用代谢工程来创建可以绕过正常代谢限制以将底物转化为氢的菌株。大肠杆菌可以将多种糖降解为氢，但只能将丙酮酸结处可用的电子转化为氢，并且不能使用糖酵解过程中产生的NADH中可用的电子。在此，来自真养罗尔斯通氏菌H16的可溶性[NiFe]氢化酶（SH氢化酶）的异源表达被用于证明能够从发酵产生的NADH获得大量氢的途径的引入。通过体外酶活性测定证明成功表达。此外，SH的表达使葡萄糖上的厌氧生长恢复为adhE菌株，该菌株通常由于不能再氧化NADH而生长受阻。体内产氢的测量结果表明，几种代谢工程菌株能够使用SH氢化酶产生2摩尔H2/摩尔葡萄糖消耗，接近理论最大值。异源[NiFe]氢化酶在E.大肠杆菌的产氢活性依赖于NAD（P）H，但产氢水平很低。在这里，我们首次显示了大量的体内氢生产的异源表达的[NiFe]氢化酶，可溶性NAD依赖H2酶的R。真养型（SH氢化酶）。这种氢化酶能够将代谢产生的NADH偶联到氢的产生，从而拯救了醇脱氢酶（adhE）突变体。这扩大了可用于产氢的代谢范围，从而可能为大大改进产氢的创造打开大门。进一步提高产量的策略应该围绕提供额外的NADH。
Fermentative hydrogen production is an attractive means for the sustainable production of this future energy carrier but is hampered by low yields. One possible solution is to create, using metabolic engineering, strains which can bypass the normal metabolic limits to substrate conversion to hydrogen. Escherichia coli can degrade a variety of sugars to hydrogen but can only convert electrons available at the pyruvate node to hydrogen, and is unable to use the electrons available in NADH generated during glycolysis. Here, the heterologous expression of the soluble [NiFe] hydrogenase from Ralstonia eutropha H16 (the SH hydrogenase) was used to demonstrate the introduction of a pathway capable of deriving substantial hydrogen from the NADH generated by fermentation. Successful expression was demonstrated by in vitro assay of enzyme activity. Moreover, expression of SH restored anaerobic growth on glucose to adhE strains, normally blocked for growth due to the inability to re-oxidize NADH. Measurement of in vivo hydrogen production showed that several metabolically engineered strains were capable of using the SH hydrogenase to derive 2 mol H2 per mol of glucose consumed, close to the theoretical maximum. Previous introduction of heterologous [NiFe] hydrogenase in E. coli led to NAD(P)H dependent activity, but hydrogen production levels were very low. Here we have shown for the first time substantial in vivo hydrogen production by a heterologously expressed [NiFe] hydrogenase, the soluble NAD-dependent H2ase of R. eutropha (SH hydrogenase). This hydrogenase was able to couple metabolically generated NADH to hydrogen production, thus rescuing an alcohol dehydrogenase (adhE) mutant. This enlarges the range of metabolism available for hydrogen production, thus potentially opening the door to the creation of greatly improved hydrogen production. Strategies for further increasing yields should revolve around making additional NADH available.
登录
查看更多内容
影响因子：
0.6
作者：
Cho, Han-Saem;Kim, Young Mi;Park, Jong Moon
通讯作者：
Park, Jong Moon
影响因子：
4.1
作者：
Kim, Jaoon Y. H.;Jo, Byung Hoon;Cha, Hyung Joon
通讯作者：
Cha, Hyung Joon
影响因子：
3.2
作者：
Burgdorf, T;van der Linden, E;Friedrich, B
通讯作者：
Friedrich, B
影响因子：
3.2
作者：
GUPTA, S;CLARK, DP
通讯作者：
CLARK, DP
DOI：
10.1007/978-1-4614-1208-3_5
发表时间：
2012-01-01
期刊：
MICROBIAL TECHNOLOGIES IN ADVANCED BIOFUELS PRODUCTION
影响因子：
--
作者：
Abo-Hashesh, Mona;Hallenbeck, Patrick C.
通讯作者：
Hallenbeck, Patrick C.