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Volume 146, Issue 4

Expression and purification of four different rhizobial acyl carrier proteins

The GenBank accession numbers for the nucleotide sequences reported in this paper are AF159244 and AF159243 for the -containing regions of 1021 and of LPR5045, respectively.

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Abstract

In rhizobia, besides the constitutive acyl carrier protein (AcpP) involved in the biosynthesis and transfer of common fatty acids, there are at least three specialized acyl carrier proteins (ACPs): (1) the flavonoid-inducible nodulation protein NodF; (2) the RkpF protein, which is required for the biosynthesis of rhizobial capsular polysaccharides; and (3) AcpXL, which transfers 27-hydroxyoctacosanoic acid to a sugar backbone during lipid A biosynthesis. Whereas the nucleotide sequences encoding the three specialized ACPs are known, only the amino acid sequence of the AcpP of was available. In this study, using reverse genetics, the genes for the constitutive AcpPs of and of were cloned and sequenced. Previously, it had been shown that NodF and RkpF can be overproduced in using the T7 polymerase expression system. Using the same system, the constitutive AcpPs of and of , together with the specialized ACP AcpXL, were overproduced and purified. All the known ACPs of rhizobia can be labelled during expression in with radioactive β-alanine added to the growth medium due to their modification with a 4’-phosphopantetheine prosthetic group. The availability of all functionally different ACPs should help to unravel how different fatty acids are targeted towards different biosynthetic pathways in one organism.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): ACP, acyl carrier protein , acpP , acpXL , acyl carrier protein , iPCR, inverse PCR and Rhizobium
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2000-04-01
2024-04-27
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Expression and purification of four different rhizobial acyl carrier proteinsThe GenBank accession numbers for the nucleotide sequences reported in this paper are AF159244 and AF159243 for the acpP-containing regions of Sinorhizobium meliloti 1021 and of Rhizobium leguminosarum LPR5045, respectively.
Microbiology 146, 839 (2000); https://doi.org/10.1099/00221287-146-4-839
/content/journal/micro/10.1099/00221287-146-4-839
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