Expression and characterization of Pantoea CO dehydrogenase to utilize CO-containing industrial waste gas for expanding the versatility of CO dehydrogenase.
Expression and characterization of Pantoea CO dehydrogenase to utilize CO-containing industrial waste gas for expanding the versatility of CO dehydrogenase.
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DOI：
10.1038/srep44323
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发表时间：
2017-03-14
影响因子：
4.6
通讯作者：
Kim YH
中科院分区：
文献类型：
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作者：
Choi ES;Min K;Kim GJ;Kwon I;Kim YH
虽然好氧一氧化碳脱氢酶（CODHs）具有广泛的应用前景，但其活性低、无异源表达等问题一直制约着其实际应用。我们首次在大肠杆菌中功能表达重组PsCODH，并通过简便的方法获得了一种高浓度的重组酶。考察了其电子受体谱、最佳条件（pH 6.5和30℃）、动力学参数（kcat为12.97 s−1，Km为0.065 mM，比活性为0.86 μ g−1）。高炉煤气（BFG）是炼钢过程中产生的一种废气，含20%的CO，作为PsCODH的底物进行了试验。与纯CO和20% CO相比，即使加入BFG，重组PsCODH的活性也分别保持了88.2%和108.4%。研究结果不仅为利用含co的工业废气作为廉价、丰富和可再生的资源提供了一个有希望的策略，而且为进一步研究以CODH为基础的co利用系统通过CO2作为中间体产生增值化学品的级联反应提供了重要信息，这将最终扩大CODH的通用性。
Although aerobic CO dehydrogenases (CODHs) might be applicable in various fields, their practical applications have been hampered by low activity and no heterologous expression. We, for the first time, could functionally express recombinant PsCODH in E. coli and obtained a highly concentrated recombinant enzyme using an easy and convenient method. Its electron acceptor spectra, optimum conditions (pH 6.5 and 30 °C), and kinetic parameters (kcat of 12.97 s−1, Km of 0.065 mM, and specific activity of 0.86 Umg−1) were examined. Blast furnace gas (BFG) containing 20% CO, which is a waste gas from the steel-making process, was tested as a substrate for PsCODH. Even with BFG, the recombinant PsCODH retained 88.2% and 108.4% activity compared with those of pure CO and 20% CO, respectively. The results provide not only a promising strategy to utilize CO-containing industrial waste gases as cheap, abundant, and renewable resources but also significant information for further studies about cascade reactions producing value-added chemicals via CO2 as an intermediate produced by a CODH-based CO-utilization system, which would ultimately expand the versatility of CODH.