Synthesis of L-2,3-diaminopropionic acid, a siderophore and antibiotic precursor.
L-2,3-diaminopropionic acid (L-Dap) is an amino acid that is a precursor of antibiotics and staphyloferrin B a siderophore produced by Staphylococcus aureus. SbnA and SbnB are encoded by the staphyloferrin B biosynthetic gene cluster and are implicated in L-Dap biosynthesis. We demonstrate here that SbnA uses PLP and substrates O-phospho-L-serine and L-glutamate to produce a metabolite N-(1-amino-1-carboxyl-2-ethyl)-glutamic acid (ACEGA). SbnB is shown to use NAD(+) to oxidatively hydrolyze ACEGA to yield α-ketoglutarate and L-Dap. Also, we describe crystal structures of SbnB in complex with NADH and ACEGA as well as with NAD(+) and α-ketoglutarate to reveal the residues required for substrate binding, oxidation, and hydrolysis. SbnA and SbnB contribute to the iron sparing response of S. aureus that enables staphyloferrin B biosynthesis in the absence of an active tricarboxylic acid cycle.
Department of Microbiology and Immunology, Life Sciences Institute, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada.
Marek J
MJ
Kobylarz
Department of Microbiology and Immunology, Life Sciences Institute, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada.
Jason C
JC
Grigg
Department of Microbiology and Immunology, Life Sciences Institute, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada.
Shin-ichi J
SJ
Takayama
Department of Microbiology and Immunology, Life Sciences Institute, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada.
Dushyant K
DK
Rai
Department of Microbiology and Immunology, The Centre for Human Immunology, University of Western Ontario, London, ON N6A 5C1, Canada.
David E
DE
Heinrichs
Department of Microbiology and Immunology, Life Sciences Institute, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada. Electronic address: michael.murphy@ubc.ca.
Michael E P
ME
Murphy
D000900:Anti-Bacterial Agents / Q000737:chemistry*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D020134:Catalytic Domain; D002951:Citrates / Q000096:biosynthesis / Q000737:chemistry; D018360:Crystallography, X-Ray; D018698:Glutamic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D006868:Hydrolysis; D007656:Ketoglutaric Acids / Q000737:chemistry / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D009243:NAD / Q000737:chemistry / Q000378:metabolism; D010768:Phosphoserine / Q000031:analogs & derivatives / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D017262:Siderophores / Q000096:biosynthesis* / Q000737:chemistry; D013211:Staphylococcus aureus / Q000201:enzymology / Q000378:metabolism*; D015091:beta-Alanine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D002951:Citrates; D007656:Ketoglutaric Acids; D011994:Recombinant Proteins; D017262:Siderophores; D009243:NAD; D015091:beta-Alanine; C083439:staphyloferrin B; D010768:Phosphoserine; D018698:Glutamic Acid; C011757:2,3-diaminopropionic acid