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Proteomics approach to decipher novel genes and enzymes characterization of a bioelectricity-generating and dye-decolorizing bacterium Proteus hauseri ZMd44

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Abstract

The first-attempt study employed a proteomics strategy for the identification of abundant proteins from a bioelectricity generation and dye decolorization bacterium Proteus hauseri ZMd44. By using the degenerated primers designed based on the peptide sequences from tandem mass spectroscopy and the whole genomics annotation of the closely associated strain, Proteus penneri ATCC 35198, the genes were successfully obtained for two full-length genes of 543 bp (laccase) and 1,086 bp (Omp F, porin) encoding to 181 amino acids and 362 amino acids, respectively. It explored laccase and NADH dehydrogenase involvement in the oxidation-reduction reaction as well, as porin played an important role in providing channels for related proteins in the accomplishment of electron transportation in P. hauseri. Detailed enzymatic assays indicated that laccase activity of 542.2 U/DCW could be stimulated by 2.5 mM copper induction in LB medium (ca. 293-fold to those without copper induction). Among intracellular proteins, NADH dehydrogenase activity of 257.2 U/mg via mediator riboflavin was in parallel with the decolorizing capability of azo dye Rb160 that only took place in LB medium. From the evaluation of kinetic parameters (Vmax and Km were 0.272 U/min and 0.393 mM with ABTS, 0.046 U/min and 43.8 μM with NADH), it is better to decipher the decolorization mechanism of ZMd44 indicating that laccase and NADH dehydrogenase played the most crucial role for azo dye decolorization.

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Authors and Affiliations

  1. Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361-005, China

    I. -Son Ng, Xuesong Zheng, Xiaoqin Chi & Yinghua Lu

  2. The Key Laboratory for Synthetic Biotechnology of Xiamen City, Xiamen University, Xiamen, 361-005, China

    I. -Son Ng & Yinghua Lu

  3. Department of Chemical and Materials Engineering, National I-Lan University, I-Lan, 26047, Taiwan

    Bor-Yann Chen

  4. Department of Biotechnology, Southern Taiwan University of Science Technology, Tainan County, 71005, Taiwan

    Chun-Sheng Chang

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  1. I. -Son Ng
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Correspondence to I. -Son Ng.

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Ng, I.S., Zheng, X., Chen, BY. et al. Proteomics approach to decipher novel genes and enzymes characterization of a bioelectricity-generating and dye-decolorizing bacterium Proteus hauseri ZMd44. Biotechnol Bioproc E 18, 8–17 (2013). https://doi.org/10.1007/s12257-012-0340-7

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  • DOI: https://doi.org/10.1007/s12257-012-0340-7

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