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Cloning and Characterization of the Genes for Biosynthesis of the Compatible Solute Ectoine in the Moderately Halophilic Bacterium Halobacillus dabanensis D-8T

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Abstract

A 11.2-kb fragment containing the ectABC genes of the biosynthetic pathway of ectoine from the Gram-positive, moderately halophilic bacterium Halobacillus dabanensis D-8T was obtained by inverse polymerase chain reaction. Subsequently, the entire ectABC cluster was cloned and analyzed. It revealed that the intergenic regions of the ectABC genes from H. dabanensis D-8T are more tightly spaced than those of Chromohalobacter salexigens, Halomonas elongata, Marinococcus halophilus, and Salibacillus pasteurii. The amino-acid sequence deduced from ectABC was highly homologous that from Virgibacillus pantethenticus (EctA 52%, EctB 60%, EctC 67%, respectively). The ectABC genes were cloned in the expression plasmid pMXB10 resulting in pMXB10ectABC. The ectoine was detected from cell extract in Escherishia coli ER2566 containing pMXB10ectABC using 13C nuclear magnetic resonance spectroscopy.

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Acknowledgments

We are grateful to T. Van Brussel (Leiden University, The Netherlands) for critical reading of the manuscript. We thank New England Biolabs for providing the plasmid pMBX10 and E. coli ER2566. This work was supported by the Chinese National Program for High Technology Research and Development (2003AA241150).

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  1. Department of Microbiology, College of Biologic Sciences, China Agricultural University and Key Laboratory of Agro-Microbial Resource and Application, Ministry of Agriculture, Beijing, 100094, PRC

    B. Zhao, W. Lu, L. Yang, B. Zhang, L. Wang & S. S. Yang

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  1. B. Zhao
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  2. W. Lu
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  3. L. Yang
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  4. B. Zhang
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  6. S. S. Yang
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Correspondence to S. S. Yang.

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Zhao, B., Lu, W., Yang, L. et al. Cloning and Characterization of the Genes for Biosynthesis of the Compatible Solute Ectoine in the Moderately Halophilic Bacterium Halobacillus dabanensis D-8T . Curr Microbiol 53, 183–188 (2006). https://doi.org/10.1007/s00284-005-0396-0

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