title: Bacterial abl-like genes: production of the archaeal osmolyte N(ε)-acetyl-β-lysine by homologous overexpression of the yodP-kamA genes in Bacillus subtilis.
abstract: Nε-acetyl-β-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(ε)-acetyl-β-lysine. The biochemical and biophysical properties of N(ε)-acetyl-β-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(ε)-acetyl-β-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(ε)-acetyl-β-lysine. The data reported here thus open the bottleneck for the large-scale production of N(ε)-acetyl-β-lysine to investigate its properties as a compatible solute.
authors.affiliation: Molecular Microbiology & Bioenergetics, Institute of Molecular Biosciences, Goethe University, Max-von-Laue-Str. 9, 60438 Frankfurt am Main, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Müller
authors.forename: Tamara
authors.initials: T
authors.lastname: Hoffmann
authors.forename: Helena
authors.initials: H
authors.lastname: Santos
authors.forename: Stephan H
authors.initials: SH
authors.lastname: Saum
authors.forename: Erhard
authors.initials: E
authors.lastname: Bremer
authors.forename: Volker
authors.initials: V
authors.lastname: Müller
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D016313:Genes, abl; D008239:Lysine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D005810:Multigene Family; D019906:Nuclear Magnetic Resonance, Biomolecular
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; C523231:yoeB protein, Bacillus subtilis; C075812:N'-acetyl-beta-lysine; D008239:Lysine