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Influence of different factors on the nitrogenase activity of the engineered Escherichia coli 78-7

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Abstract

The engineered Escherichia coli 78-7 is a derivative of E. coli JM109 carrying a nitrogen fixation (nif) gene cluster composed of nine genes (nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV) and its own σ70-dependent nif promoter from a gram-positive bacterium Paenibacillus sp. WLY78. The physiological and biochemical characteristics of the engineered E. coli 78-7 were analyzed by using Biolog GEN III MicroPlate, with E. coli JM109 and JM109/pHY300PLK (E. coli JM109 carrying empty vector) as controls. Analysis of 94 phenotypic tests: 71 carbon source utilization assays and 23 chemical sensitivity tests showed that the engineered E. coli 78-7, E. coli JM109 and JM109/pHY300PLK gave similar patterns of utilization of various substrates as carbon and energy sources. Furthermore, the effect of carbon source, nitrogen source, culture temperature on the nitrogenase activity of the engineered E. coli 78-7 was investigated. Our study demonstrates that the nif capacity of E. coli 78-7 was affected significantly by the different culture condition. The significant nitrogenase activity of E. coli 78-7 were obtained when cells were cultivated in the medium containing 4 g/l glucose (carbon source) and 2 mM glutamate (nitrogen source) and at 30 °C.

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Acknowledgments

This work was supported by Guangdong Innovative and Entrepreneurial Research Team Program (No. 2013S033) and from the National High Technology Research and Development Program (“863” Program No. 2013AA102802-04).

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Authors and Affiliations

  1. State Key Laboratory for Agrobiotechnology and College of Biological Sciences, China Agricultural University, Yuanmingyuan Road, Haidian District, Beijing, 100193, China

    Li-hong Zhang & San-feng Chen

  2. College of Life Science, Shanxi Normal University, Linfen, 041000, ShanXi, China

    Li-hong Zhang

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  1. Li-hong Zhang
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  2. San-feng Chen
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Correspondence to San-feng Chen.

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Zhang, Lh., Chen, Sf. Influence of different factors on the nitrogenase activity of the engineered Escherichia coli 78-7. World J Microbiol Biotechnol 31, 921–927 (2015). https://doi.org/10.1007/s11274-015-1846-x

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  • DOI: https://doi.org/10.1007/s11274-015-1846-x

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