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Bacterial abl-like genes: production of the archaeal osmolyte \( {N^{\varepsilon }}{\text{ - acetyl - }}\beta {\text{ - lysine}} \) by homologous overexpression of the yodP–kamA genes in Bacillus subtilis

  • Applied Genetics and Molecular Biotechnology
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Abstract

\( {N^{\varepsilon }}{\text{ - acetyl - }}\beta {\text{ - lysine}} \) is an archaeal compatible solute whose synthesis is mediated by the sequential reactions of the lysine-2,3-aminomutase AblA and the acetyltransferase AblB. α-Lysine serves as the precursor and is converted by AblA to β-lysine, and AblB then acetylates this intermediate to \( {N^{\varepsilon }}{\text{ - acetyl - }}\beta {\text{ - lysine}} \). The biochemical and biophysical properties of \( {N^{\varepsilon }}{\text{ - acetyl - }}\beta {\text{ - lysine}} \) have so far not been studied intensively due to restrictions in the supply of this compound. A search for ablAB-like genes in the genomes of members of the family Bacillaceae revealed the yodP–kamA genes that encode a AblA-related lysine-2,3-aminomutase and AblB-related putative acetyltransferase. In Bacillus subtilis, the yodP–kamA genes are part of a transcriptional unit (yodT–yodS–yodR–yodQ–yodP–kamA) whose expression is upregulated during sporulation and controlled by the mother-cell-specific transcription factor SigE. \( {N^{\varepsilon }}{\text{ - acetyl - }}\beta {\text{ - lysine}} \) was not detectable in vegetatively growing or osmotically stressed B. subtilis cells, and the deletion of the yodT–yodS–yodR–yodQ–yodP–kamA region had no noticeable effects on growth in rich or minimal media or osmotic stress resistance. However, when we expressed the yodP–kamA genes outside their natural genetic context from an isopropyl β-d-1-thiogalactopyranoside-inducible promoter on a plasmid in B. subtilis, the recombinant strain synthesized considerable amounts (0.28 μmol/mg protein) of \( {N^{\varepsilon }}{\text{ - acetyl - }}\beta {\text{ - lysine}} \). The data reported here thus open the bottleneck for the large-scale production of \( {N^{\varepsilon }}{\text{ - acetyl - }}\beta {\text{ - lysine}} \) to investigate its properties as a compatible solute.

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Acknowledgements

This work was supported by a grant from the Deutsche Forschungsgemeinschaft (to V.M.) and by a grant through the LOEWE initiative of the State of Hessen (SynMicro LOEWE Centre; Marburg) (to E.B). The NMR spectrometers are part of the National NMR Network (REDE/1517/RMN/2005), supported by Programa Operacional Ciência e Inovação 2010 and Fundação para a Ciência e a Tecnologia.

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  1. Stephan H. Saum

    Present address: Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Avenue, Boston, MA, 02115, USA

Authors and Affiliations

  1. Molecular Microbiology & Bioenergetics, Institute of Molecular Biosciences, Goethe University, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany

    Stefan Müller, Stephan H. Saum & Volker Müller

  2. Laboratorium für Mikrobiologie, Philipps Universität Marburg, Karl-von-Frisch Str. 8, 35043, Marburg, Germany

    Tamara Hoffmann & Erhard Bremer

  3. Instituto de Tecnologia Química e Biológica, Universida de Nova de Lisboa, 2780-156, Oeiras, Portugal

    Helena Santos

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Correspondence to Volker Müller.

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Suppl. Table 1

Identity and homology of B. subtilis YodP to corresponding enzymes in other organisms. (DOC 13 kb)

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Müller, S., Hoffmann, T., Santos, H. et al. Bacterial abl-like genes: production of the archaeal osmolyte \( {N^{\varepsilon }}{\text{ - acetyl - }}\beta {\text{ - lysine}} \) by homologous overexpression of the yodP–kamA genes in Bacillus subtilis . Appl Microbiol Biotechnol 91, 689–697 (2011). https://doi.org/10.1007/s00253-011-3301-8

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