1887

Abstract

Biotin is a key cofactor of metabolic carboxylases, although many rhizobial strains are biotin auxotrophs. When some of these strains were serially subcultured in minimal medium, they showed diminished growth and increased excretion of metabolites. The addition of biotin, or genetic complementation with biotin synthesis genes resulted in full growth of CFN42 and CIAT652 strains. Half of rhizobial genomes did not show genes for biotin biosynthesis, but three-quarters had genes for biotin transport. Some strains had genes for an avidin homologue (rhizavidin), a protein with high affinity for biotin but an unknown role in bacteria. A CFN42-derived rhizavidin mutant showed a sharper growth decrease in subcultures, revealing a role in biotin storage. In the search of biotin-independent growth of subcultures, CFN42 and CIAT652 strains with excess aeration showed optimal growth, as they also did, unexpectedly, with the addition of aspartic acid analogues α- and -methyl aspartate. Aspartate analogues can be sensed by the chemotaxis aspartate receptor Tar. A homologue was identified and its mutants showed no growth recovery with aspartate analogues, indicating requirement of the Tar receptor in such a phenotype. Additionally, mutants did not recover full growth with excess aeration. A Rubisco-like protein was found to be necessary for growth as the corresponding mutants showed no recovery either with high aeration or aspartate analogues; also, diminished carboxylation was observed. Taken together, our results indicate a route of biotin-independent growth in rhizobial strains that included oxygen, a Tar receptor and a previously uncharacterized Rubisco-like protein.

Funding
This study was supported by the:
  • Dirección General de Asuntos del Personal Académico, Universidad Nacional Autónoma de México (Award PAPIIT IN201617, PAPIIT IN204320)
    • Principle Award Recipient: LourdesGirard
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2022-01-25
2024-04-20
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References

  1. Masson-Boivin C, Sachs JL. Symbiotic nitrogen fixation by rhizobia-the roots of a success story. Curr Opin Plant Biol 2018; 44:7–15 [View Article] [PubMed]
    [Google Scholar]
  2. Encarnación S, Dunn M, Willms K, Mora J. Fermentative and aerobic metabolism in Rhizobium etli. J Bacteriol 1995; 177:3058–3066 [View Article] [PubMed]
    [Google Scholar]
  3. Dunn MF, Encarnación S, Araíza G, Vargas MC, Dávalos A et al. Pyruvate carboxylase from Rhizobium etli: mutant characterization, nucleotide sequence, and physiological role. J Bacteriol 1996; 178:5960–5970 [View Article] [PubMed]
    [Google Scholar]
  4. Dunn MF, Araíza G, Mora J. Biochemical characterization of a Rhizobium etli monovalent cation-stimulated acyl-coenzyme A carboxylase with a high substrate specificity constant for propionyl-coenzyme A. Microbiology (Reading) 2004; 150:399–406 [View Article] [PubMed]
    [Google Scholar]
  5. Tong L. Structure and function of biotin-dependent carboxylases. Cell Mol Life Sci 2013; 70:863–891 [View Article] [PubMed]
    [Google Scholar]
  6. Yokota A. Revisiting RuBisCO. Biosci Biotechnol Biochem 2017; 81:2039–2049 [View Article] [PubMed]
    [Google Scholar]
  7. Ashida H, Saito Y, Kojima C, Kobayashi K, Ogasawara N et al. A functional link between RuBisCO-like protein of Bacillus and photosynthetic RuBisCO. Science 2003; 302:286–290 [View Article] [PubMed]
    [Google Scholar]
  8. Hanson TE, Tabita FR. Insights into the stress response and sulfur metabolism revealed by proteome analysis of a Chlorobium tepidum mutant lacking the Rubisco-like protein. Photosynthesis Research 2003; 78:231–248 [View Article]
    [Google Scholar]
  9. Erb TJ, Evans BS, Cho K, Warlick BP, Sriram J et al. A RubisCO-like protein links SAM metabolism with isoprenoid biosynthesis. Nat Chem Biol 2012; 8:926–932 [View Article] [PubMed]
    [Google Scholar]
  10. Bi S, Sourjik V. Stimulus sensing and signal processing in bacterial chemotaxis. Curr Opin Microbiol 2018; 45:22–29 [View Article] [PubMed]
    [Google Scholar]
  11. Götz R, Schmitt R. Rhizobium meliloti swims by unidirectional, intermittent rotation of right-handed flagellar helices. J Bacteriol 1987; 169:3146–3150 [View Article] [PubMed]
    [Google Scholar]
  12. Alanis-Sánchez BM, Pérez-Tapia SM, Vázquez-Leyva S, Mejía-Calvo I, Macías-Palacios Z et al. Utilization of naproxen by Amycolatopsis sp. Poz 14 and detection of the enzymes involved in the degradation metabolic pathway. World J Microbiol Biotechnol 2019; 35:186 [View Article] [PubMed]
    [Google Scholar]
  13. Meier VM, Muschler P, Scharf BE. Functional analysis of nine putative chemoreceptor proteins in Sinorhizobium meliloti. J Bacteriol 2007; 189:1816–1826 [View Article] [PubMed]
    [Google Scholar]
  14. Caetano-Anollés G, Wall LG, De Micheli AT, Macchi EM, Bauer WD et al. Role of motility and chemotaxis in efficiency of nodulation by Rhizobium meliloti. Plant Physiol 1988; 86:1228–1235 [View Article] [PubMed]
    [Google Scholar]
  15. Yost CK, Del Bel KL, Quandt J, Hynes MF. Rhizobium leguminosarum methyl-accepting chemotaxis protein genes are down-regulated in the pea nodule. Arch Microbiol 2004; 182:505–513 [View Article] [PubMed]
    [Google Scholar]
  16. Yost CK, Rochepeau P, Hynes MF. Rhizobium leguminosarum contains a group of genes that appear to code for methyl-accepting chemotaxis proteins. Microbiology (Reading) 1998; 144 (Pt 7):1945–1956 [View Article] [PubMed]
    [Google Scholar]
  17. Van Bastelaere E, Lambrecht M, Vermeiren H, Van Dommelen A, Keijers V et al. Characterization of a sugar-binding protein from Azospirillum brasilense mediating chemotaxis to and uptake of sugars. Mol Microbiol 1999; 32:703–714 [View Article] [PubMed]
    [Google Scholar]
  18. Mora Y, Díaz R, Vargas-Lagunas C, Peralta H, Guerrero G et al. Nitrogen-fixing rhizobial strains isolated from common bean seeds: phylogeny, physiology, and genome analysis. Appl Environ Microbiol 2014; 80:5644–5654 [View Article]
    [Google Scholar]
  19. Helppolainen SH, Nurminen KP, Määttä JAE, Halling KK, Slotte JP et al. Rhizavidin from Rhizobium etli: the first natural dimer in the avidin protein family. Biochem J 2007; 405:397–405 [View Article] [PubMed]
    [Google Scholar]
  20. Shevchuk NA, Bryksin AV, Nusinovich YA, Cabello FC, Sutherland M et al. Construction of long DNA molecules using long PCR-based fusion of several fragments simultaneously. Nucleic Acids Res 2004; 32:e19 [View Article] [PubMed]
    [Google Scholar]
  21. Schäfer A, Tauch A, Jäger W, Kalinowski J, Thierbach G et al. Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene 1994; 145:69–73 [View Article] [PubMed]
    [Google Scholar]
  22. Martinez-Salazar JM, Romero D. Role of the ruvB gene in homologous and homeologous recombination in Rhizobium etli. Gene 2000; 243:125–131 [View Article] [PubMed]
    [Google Scholar]
  23. Hynes MF, McGregor NF. Two plasmids other than the nodulation plasmid are necessary for formation of nitrogen-fixing nodules by Rhizobium leguminosarum. Mol Microbiol 1990; 4:567–574 [View Article] [PubMed]
    [Google Scholar]
  24. Sullivan JT, Brown SD, Yocum RR, Ronson CW. The bio operon on the acquired symbiosis island of Mesorhizobium sp. strain R7A includes a novel gene involved in pimeloyl-CoA synthesis. Microbiology (Reading) 2001; 147:1315–1322 [View Article] [PubMed]
    [Google Scholar]
  25. Quinto C, De La Vega H, Flores M, Leemans J, Cevallos MA et al. Nitrogenase reductase: A functional multigene family in Rhizobium phaseoli. Proc Natl Acad Sci U S A 1985; 82:1170–1174 [View Article] [PubMed]
    [Google Scholar]
  26. González V, Acosta JL, Santamaría RI, Bustos P, Fernández JL et al. Conserved symbiotic plasmid DNA sequences in the multireplicon pangenomic structure of Rhizobium etli. Appl Environ Microbiol 2010; 76:1604–1614 [View Article]
    [Google Scholar]
  27. Nordlund HR, Hytönen VP, Laitinen OH, Kulomaa MS. Novel avidin-like protein from a root nodule symbiotic bacterium, Bradyrhizobium japonicum. J Biol Chem 2005; 280:13250–13255 [View Article] [PubMed]
    [Google Scholar]
  28. Meir A, Helppolainen SH, Podoly E, Nordlund HR, Hytönen VP et al. Crystal structure of rhizavidin: insights into the enigmatic high-affinity interaction of an innate biotin-binding protein dimer. J Mol Biol 2009; 386:379–390 [View Article] [PubMed]
    [Google Scholar]
  29. Guillén-Navarro K, Encarnación S, Dunn MF. Biotin biosynthesis, transport and utilization in rhizobia. FEMS Microbiol Lett 2005; 246:159–165 [View Article] [PubMed]
    [Google Scholar]
  30. Helliwell KE, Collins S, Kazamia E, Purton S, Wheeler GL et al. Fundamental shift in vitamin B12 eco-physiology of a model alga demonstrated by experimental evolution. ISME J 2015; 9:1446–1455 [View Article] [PubMed]
    [Google Scholar]
  31. Merhej V, Royer-Carenzi M, Pontarotti P, Raoult D. Massive comparative genomic analysis reveals convergent evolution of specialized bacteria. Biol Direct 2009; 4:13 [View Article] [PubMed]
    [Google Scholar]
  32. White 3rd HB Biotin-binding proteins and biotin transport to oocytes. Ann N Y Acad Sci 1985; 447:202–211 [View Article] [PubMed]
    [Google Scholar]
  33. Hytonen VP. (Strept)avidin as a template for ligands other than biotin: an overview. Meth Enzymol 2020; 633:21–28
    [Google Scholar]
  34. Kramer KJ, Morgan TD, Throne JE, Dowell FE, Bailey M et al. Transgenic avidin maize is resistant to storage insect pests. Nat Biotechnol 2000; 18:670–674 [View Article] [PubMed]
    [Google Scholar]
  35. Adler J. Chemoreceptors in bacteria. Science 1969; 166:1588–1597 [View Article] [PubMed]
    [Google Scholar]
  36. Mesibov R, Adler J. Chemotaxis toward amino acids in Escherichia coli. J Bacteriol 1972; 112:315–326 [View Article] [PubMed]
    [Google Scholar]
  37. Parales RE, Luu RA, Chen GY, Liu X, Wu V et al. Pseudomonas putida F1 has multiple chemoreceptors with overlapping specificity for organic acids. Microbiology 2013; 159:1086–1096 [View Article]
    [Google Scholar]
  38. Xu N, Wang M, Yang X, Xu Y, Guo M. In silico analysis of the chemotactic system of Agrobacterium tumefaciens. Microb Genom 2020; 6:mgen000460 [View Article] [PubMed]
    [Google Scholar]
  39. Tabita FR, Hanson TE, Li H, Satagopan S, Singh J et al. Function, structure, and evolution of the RubisCO-like proteins and their RubisCO homologs. Microbiol Mol Biol Rev 2007; 71:576–599 [View Article] [PubMed]
    [Google Scholar]
  40. Kim SM, Lim HS, Lee SB. Discovery of a RuBisCO-like protein that functions as an oxygenase in the novel d-hamamelose pathway. Biotechnol Bioproc E 2018; 23:490–499 [View Article]
    [Google Scholar]
  41. Noel KD, Sanchez A, Fernandez L, Leemans J, Cevallos MA. Rhizobium phaseoli symbiotic mutants with transposon Tn5 insertions. J Bacteriol 1984; 158:148–155 [View Article] [PubMed]
    [Google Scholar]
  42. Quandt J, Hynes MF. Versatile suicide vectors which allow direct selection for gene replacement in gram-negative bacteria. Gene 1993; 127:15–21 [View Article] [PubMed]
    [Google Scholar]
  43. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA et al. Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene 1995; 166:175–176 [View Article] [PubMed]
    [Google Scholar]
  44. Figurski DH, Helinski DR. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A 1979; 76:1648–1652 [View Article] [PubMed]
    [Google Scholar]
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