Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Feb;179(4):1013–1022. doi: 10.1128/jb.179.4.1013-1022.1997

Genetic characterization of the pdu operon: use of 1,2-propanediol in Salmonella typhimurium.

D Walter 1, M Ailion 1, J Roth 1
PMCID: PMC178792  PMID: 9023178

Abstract

Salmonella typhimurium is able to catabolize 1,2-propanediol for use as the sole carbon and energy source; the first enzyme of this pathway requires the cofactor adenosyl cobalamin (Ado-B12). Surprisingly, Salmonella can use propanediol as the sole carbon source only in the presence of oxygen but can synthesize Ado-B12 only anaerobically. To understand this situation, we have studied the pdu operon, which encodes proteins for propanediol degradation. A set of pdu mutants defective in aerobic degradation of propanediol (with exogenous vitamin B12) defines four distinct complementation groups. Mutations in two of these groups (pduC and pduD) eliminate propanediol dehydratase activity. Based on mutant phenotypes, a third complementation group (pduG) appears to encode a cobalamin adenosyl transferase activity. No function has been assigned to the pduJ complementation group. Propionaldehyde dehydrogenase activity is eliminated by mutations in any of the four identified complementation groups, suggesting that this activity may require a complex of proteins encoded by the operon. None of the mutations analyzed affects either of the first two genes of the operon (pduA and pduB), which were identified by DNA sequence analysis. Available data suggest that the pdu operon includes enough DNA for about 15 genes and that the four genetically identified genes are the only ones required for aerobic use of propanediol.

Full Text

The Full Text of this article is available as a PDF (165.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ailion M., Bobik T. A., Roth J. R. Two global regulatory systems (Crp and Arc) control the cobalamin/propanediol regulon of Salmonella typhimurium. J Bacteriol. 1993 Nov;175(22):7200–7208. doi: 10.1128/jb.175.22.7200-7208.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berkowitz D., Hushon J. M., Whitfield H. J., Jr, Roth J., Ames B. N. Procedure for identifying nonsense mutations. J Bacteriol. 1968 Jul;96(1):215–220. doi: 10.1128/jb.96.1.215-220.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bobik T. A., Ailion M., Roth J. R. A single regulatory gene integrates control of vitamin B12 synthesis and propanediol degradation. J Bacteriol. 1992 Apr;174(7):2253–2266. doi: 10.1128/jb.174.7.2253-2266.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bochner B. R., Huang H. C., Schieven G. L., Ames B. N. Positive selection for loss of tetracycline resistance. J Bacteriol. 1980 Aug;143(2):926–933. doi: 10.1128/jb.143.2.926-933.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Castilho B. A., Olfson P., Casadaban M. J. Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons. J Bacteriol. 1984 May;158(2):488–495. doi: 10.1128/jb.158.2.488-495.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chen P., Andersson D. I., Roth J. R. The control region of the pdu/cob regulon in Salmonella typhimurium. J Bacteriol. 1994 Sep;176(17):5474–5482. doi: 10.1128/jb.176.17.5474-5482.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cunningham P. R., Clark D. P. The use of suicide substrates to select mutants of Escherichia coli lacking enzymes of alcohol fermentation. Mol Gen Genet. 1986 Dec;205(3):487–493. doi: 10.1007/BF00338087. [DOI] [PubMed] [Google Scholar]
  8. Elliott T., Roth J. R. Characterization of Tn10d-Cam: a transposition-defective Tn10 specifying chloramphenicol resistance. Mol Gen Genet. 1988 Aug;213(2-3):332–338. doi: 10.1007/BF00339599. [DOI] [PubMed] [Google Scholar]
  9. Escalante-Semerena J. C., Suh S. J., Roth J. R. cobA function is required for both de novo cobalamin biosynthesis and assimilation of exogenous corrinoids in Salmonella typhimurium. J Bacteriol. 1990 Jan;172(1):273–280. doi: 10.1128/jb.172.1.273-280.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Evans C. T., Sumegi B., Srere P. A., Sherry A. D., Malloy C. R. [13C]propionate oxidation in wild-type and citrate synthase mutant Escherichia coli: evidence for multiple pathways of propionate utilization. Biochem J. 1993 May 1;291(Pt 3):927–932. doi: 10.1042/bj2910927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fernández-Briera A., Garrido-Pertierra A. A degradation pathway of propionate in Salmonella typhimurium LT-2. Biochimie. 1988 Jun;70(6):757–768. doi: 10.1016/0300-9084(88)90105-8. [DOI] [PubMed] [Google Scholar]
  12. Hughes K. T., Roth J. R., Olivera B. M. A genetic characterization of the nadC gene of Salmonella typhimurium. Genetics. 1991 Apr;127(4):657–670. doi: 10.1093/genetics/127.4.657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Iuchi S., Chepuri V., Fu H. A., Gennis R. B., Lin E. C. Requirement for terminal cytochromes in generation of the aerobic signal for the arc regulatory system in Escherichia coli: study utilizing deletions and lac fusions of cyo and cyd. J Bacteriol. 1990 Oct;172(10):6020–6025. doi: 10.1128/jb.172.10.6020-6025.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jeter R. M. Cobalamin-dependent 1,2-propanediol utilization by Salmonella typhimurium. J Gen Microbiol. 1990 May;136(5):887–896. doi: 10.1099/00221287-136-5-887. [DOI] [PubMed] [Google Scholar]
  15. Jeter R. M., Olivera B. M., Roth J. R. Salmonella typhimurium synthesizes cobalamin (vitamin B12) de novo under anaerobic growth conditions. J Bacteriol. 1984 Jul;159(1):206–213. doi: 10.1128/jb.159.1.206-213.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kleckner N., Bender J., Gottesman S. Uses of transposons with emphasis on Tn10. Methods Enzymol. 1991;204:139–180. doi: 10.1016/0076-6879(91)04009-d. [DOI] [PubMed] [Google Scholar]
  17. LeVine S. M., Ardeshir F., Ames G. F. Isolation and Characterization of acetate kinase and phosphotransacetylase mutants of Escherichia coli and Salmonella typhimurium. J Bacteriol. 1980 Aug;143(2):1081–1085. doi: 10.1128/jb.143.2.1081-1085.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Obradors N., Badía J., Baldomà L., Aguilar J. Anaerobic metabolism of the L-rhamnose fermentation product 1,2-propanediol in Salmonella typhimurium. J Bacteriol. 1988 May;170(5):2159–2162. doi: 10.1128/jb.170.5.2159-2162.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Price G. D., Howitt S. M., Harrison K., Badger M. R. Analysis of a genomic DNA region from the cyanobacterium Synechococcus sp. strain PCC7942 involved in carboxysome assembly and function. J Bacteriol. 1993 May;175(10):2871–2879. doi: 10.1128/jb.175.10.2871-2879.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rondon M. R., Escalante-Semerena J. C. The poc locus is required for 1,2-propanediol-dependent transcription of the cobalamin biosynthetic (cob) and propanediol utilization (pdu) genes of Salmonella typhimurium. J Bacteriol. 1992 Apr;174(7):2267–2272. doi: 10.1128/jb.174.7.2267-2272.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Roof D. M., Roth J. R. Functions required for vitamin B12-dependent ethanolamine utilization in Salmonella typhimurium. J Bacteriol. 1989 Jun;171(6):3316–3323. doi: 10.1128/jb.171.6.3316-3323.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Roy I., Leadlay P. F. Physical map location of the new Escherichia coli gene sbm. J Bacteriol. 1992 Sep;174(17):5763–5764. doi: 10.1128/jb.174.17.5763-5764.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schmieger H. A method for detection of phage mutants with altered transducing ability. Mol Gen Genet. 1971;110(4):378–381. doi: 10.1007/BF00438281. [DOI] [PubMed] [Google Scholar]
  25. Stojiljkovic I., Bäumler A. J., Heffron F. Ethanolamine utilization in Salmonella typhimurium: nucleotide sequence, protein expression, and mutational analysis of the cchA cchB eutE eutJ eutG eutH gene cluster. J Bacteriol. 1995 Mar;177(5):1357–1366. doi: 10.1128/jb.177.5.1357-1366.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Suh S., Escalante-Semerena J. C. Purification and initial characterization of the ATP:corrinoid adenosyltransferase encoded by the cobA gene of Salmonella typhimurium. J Bacteriol. 1995 Feb;177(4):921–925. doi: 10.1128/jb.177.4.921-925.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tobimatsu T., Hara T., Sakaguchi M., Kishimoto Y., Wada Y., Isoda M., Sakai T., Toraya T. Molecular cloning, sequencing, and expression of the genes encoding adenosylcobalamin-dependent diol dehydrase of Klebsiella oxytoca. J Biol Chem. 1995 Mar 31;270(13):7142–7148. doi: 10.1074/jbc.270.13.7142. [DOI] [PubMed] [Google Scholar]
  28. Toraya T., Honda S., Fukui S. Fermentation of 1,2-propanediol with 1,2-ethanediol by some genera of Enterobacteriaceae, involving coenzyme B12-dependent diol dehydratase. J Bacteriol. 1979 Jul;139(1):39–47. doi: 10.1128/jb.139.1.39-47.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Toraya T., Ushio K., Fukui S., Hogenkamp P. C. Studies on the mechanism of the adenosylcobalamin-dependent diol dehydrase reaction by the use of analogs of the coenzyme. J Biol Chem. 1977 Feb 10;252(3):963–970. [PubMed] [Google Scholar]
  30. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  31. Way J. C., Davis M. A., Morisato D., Roberts D. E., Kleckner N. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene. 1984 Dec;32(3):369–379. doi: 10.1016/0378-1119(84)90012-x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES