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Artificial citrate operon and Vitreoscilla hemoglobin gene enhanced mineral phosphate solubilizing ability of Enterobacter hormaechei DHRSS

  • Applied microbial and cell physiology
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Abstract

Mineral phosphate solubilization by bacteria is mediated through secretion of organic acids, among which citrate is one of the most effective. To overproduce citrate in bacterial systems, an artificial citrate operon comprising of genes encoding NADH-insensitive citrate synthase of E. coli and Salmonella typhimurium sodium-dependent citrate transporter was constructed. In order to improve its mineral phosphate solubilizing (MPS) ability, the citrate operon was incorporated into E. hormaechei DHRSS. The artificial citrate operon transformant secreted 7.2 mM citric acid whereas in the native strain, it was undetectable. The transformant released 0.82 mM phosphate in flask studies in buffered medium containing rock phosphate as sole P source. In fermenter studies, similar phenotype was observed under aerobic conditions. However, under microaerobic conditions, no citrate was detected and P release was not observed. Therefore, an artificial citrate gene cluster containing Vitreoscilla hemoglobin (vgb) gene under its native promoter, along with artificial citrate operon under constitutive tac promoter, was constructed and transformed into E. hormaechei DHRSS. This transformant secreted 9 mM citric acid under microaerobic conditions and released 1.0 mM P. Thus, incorporation of citrate operon along with vgb gene improves MPS ability of E. hormaechei DHRSS under buffered, microaerobic conditions mimicking rhizospheric environment.

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Acknowledgments

We thank Dr. Hemanta Adhikary for providing pUCPM18-YF and pBKS-citC plasmid. CK was provided with fellowship from Council for Scientific and Industrial Research (CSIR), New Delhi, Government of India.

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Authors and Affiliations

  1. Molecular Microbial Biochemistry Laboratory, Department of Biochemistry, Faculty of Science, The M. S. University of Baroda, Vadodara, 390002, Gujarat, India

    Kavita Yadav, Chanchal Kumar & G. Naresh Kumar

  2. Department of Microbiology and Biotechnology Center, Faculty of Science, The M. S. University of Baroda, Vadodara, 390002, Gujarat, India

    G. Archana

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  1. Kavita Yadav
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  2. Chanchal Kumar
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  3. G. Archana
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  4. G. Naresh Kumar
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Correspondence to G. Naresh Kumar.

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Yadav, K., Kumar, C., Archana, G. et al. Artificial citrate operon and Vitreoscilla hemoglobin gene enhanced mineral phosphate solubilizing ability of Enterobacter hormaechei DHRSS. Appl Microbiol Biotechnol 98, 8327–8336 (2014). https://doi.org/10.1007/s00253-014-5912-3

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  • DOI: https://doi.org/10.1007/s00253-014-5912-3

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