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. 1996 Aug;178(16):4814–4821. doi: 10.1128/jb.178.16.4814-4821.1996

Isolation and nucleotide sequence of the GDP-mannose:cellobiosyl-diphosphopolyprenol alpha-mannosyltransferase gene from Acetobacter xylinum.

E A Petroni 1, L Ielpi 1
PMCID: PMC178262  PMID: 8759843

Abstract

A genetic locus from Acetobacter xylinum involved in acetan polysaccharide synthesis has been characterized. The chromosomal region was identified by screening a genomic library of A. xylinum in a Xanthomonas campestris mutant defective in xanthan polysaccharide synthesis. The A. xylinum cosmid clone can functionally complement a xanthan-negative mutant. The polymer produced by the recombinant strain was found to be indistinguishable from xanthan. Insertion mutagenesis and subcloning of the cosmid clone combined with complementation studies allowed the identification of a 2.3-kb fragment of A. xylinum chromosomal DNA. The nucleotide sequence of this fragment was analyzed and found to contain an open reading frame (aceA) of 1,182 bp encoding a protein of 43.2 kDa. Results from biochemical and genetic analyses strongly suggest that the aceA gene encodes the GDP-mannose:cellobiosyl-diphosphopolyprenol alpha-mannosyltransferase enzyme, which is responsible for the transfer of an alpha-mannosyl residue from GDP-Man to cellobiosyl-diphosphopolyprenol. A search for similarities with other known mannosyltransferases revealed that all bacterial alpha-mannosyltransferases have a short COOH-terminal amino acid sequence in common.

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Selected References

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  1. Boulnois G. J., Roberts I. S. Genetics of capsular polysaccharide production in bacteria. Curr Top Microbiol Immunol. 1990;150:1–18. doi: 10.1007/978-3-642-74694-9_1. [DOI] [PubMed] [Google Scholar]
  2. Brede G., Fjaervik E., Valla S. Nucleotide sequence and expression analysis of the Acetobacter xylinum uridine diphosphoglucose pyrophosphorylase gene. J Bacteriol. 1991 Nov;173(21):7042–7045. doi: 10.1128/jb.173.21.7042-7045.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buendia A. M., Enenkel B., Köplin R., Niehaus K., Arnold W., Pühler A. The Rhizobium meliloti exoZl exoB fragment of megaplasmid 2: ExoB functions as a UDP-glucose 4-epimerase and ExoZ shows homology to NodX of Rhizobium leguminosarum biovar viciae strain TOM. Mol Microbiol. 1991 Jun;5(6):1519–1530. doi: 10.1111/j.1365-2958.1991.tb00799.x. [DOI] [PubMed] [Google Scholar]
  4. Cangelosi G. A., Martinetti G., Leigh J. A., Lee C. C., Thienes C., Theines C., Nester E. W. Role for [corrected] Agrobacterium tumefaciens ChvA protein in export of beta-1,2-glucan. J Bacteriol. 1989 Mar;171(3):1609–1615. doi: 10.1128/jb.171.3.1609-1615.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Coucheron D. H. A family of IS1031 elements in the genome of Acetobacter xylinum: nucleotide sequences and strain distribution. Mol Microbiol. 1993 Jul;9(1):211–218. doi: 10.1111/j.1365-2958.1993.tb01682.x. [DOI] [PubMed] [Google Scholar]
  6. Couso R. O., Ielpi L., Garcia R. C., Dankert M. A. Biosynthesis of polysaccharides in Acetobacter xylinum. Sequential synthesis of a heptasaccharide diphosphate prenol. Eur J Biochem. 1982 Apr;123(3):617–627. doi: 10.1111/j.1432-1033.1982.tb06577.x. [DOI] [PubMed] [Google Scholar]
  7. Ditta G., Schmidhauser T., Yakobson E., Lu P., Liang X. W., Finlay D. R., Guiney D., Helinski D. R. Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. Plasmid. 1985 Mar;13(2):149–153. doi: 10.1016/0147-619x(85)90068-x. [DOI] [PubMed] [Google Scholar]
  8. Fath M. J., Kolter R. ABC transporters: bacterial exporters. Microbiol Rev. 1993 Dec;57(4):995–1017. doi: 10.1128/mr.57.4.995-1017.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Glucksmann M. A., Reuber T. L., Walker G. C. Genes needed for the modification, polymerization, export, and processing of succinoglycan by Rhizobium meliloti: a model for succinoglycan biosynthesis. J Bacteriol. 1993 Nov;175(21):7045–7055. doi: 10.1128/jb.175.21.7045-7055.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gosselé F., Swings J., Mossel D. A., de Ley J. Identification of Acetobacter strains isolated from spoiled lactic acid fermented meat food for pets. Antonie Van Leeuwenhoek. 1984;50(3):269–274. doi: 10.1007/BF02342137. [DOI] [PubMed] [Google Scholar]
  11. Gramajo H. C., Viale A. M., de Mendoza D. Expression of cloned genes by in vivo insertion of tac promoter using a mini-Mu bacteriophage. Gene. 1988 May 30;65(2):305–314. doi: 10.1016/0378-1119(88)90467-2. [DOI] [PubMed] [Google Scholar]
  12. Harding N. E., Cleary J. M., Cabañas D. K., Rosen I. G., Kang K. S. Genetic and physical analyses of a cluster of genes essential for xanthan gum biosynthesis in Xanthomonas campestris. J Bacteriol. 1987 Jun;169(6):2854–2861. doi: 10.1128/jb.169.6.2854-2861.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hull R. A., Gill R. E., Hsu P., Minshew B. H., Falkow S. Construction and expression of recombinant plasmids encoding type 1 or D-mannose-resistant pili from a urinary tract infection Escherichia coli isolate. Infect Immun. 1981 Sep;33(3):933–938. doi: 10.1128/iai.33.3.933-938.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ielpi L., Couso R. O., Dankert M. A. Sequential assembly and polymerization of the polyprenol-linked pentasaccharide repeating unit of the xanthan polysaccharide in Xanthomonas campestris. J Bacteriol. 1993 May;175(9):2490–2500. doi: 10.1128/jb.175.9.2490-2500.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Iversen T., Standal R., Pedersen T., Coucheron D. H. IS1032 from Acetobacter xylinum, a new mobile insertion sequence. Plasmid. 1994 Jul;32(1):46–54. doi: 10.1006/plas.1994.1043. [DOI] [PubMed] [Google Scholar]
  16. Jann K., Jann B. Biochemistry and expression of bacterial capsules. Biochem Soc Trans. 1991 Aug;19(3):623–628. doi: 10.1042/bst0190623. [DOI] [PubMed] [Google Scholar]
  17. Jansson P. E., Kenne L., Lindberg B. Structure of extracellular polysaccharide from Xanthomonas campestris. Carbohydr Res. 1975 Dec;45:275–282. doi: 10.1016/s0008-6215(00)85885-1. [DOI] [PubMed] [Google Scholar]
  18. Jansson P. E., Lindberg J., Wimalasiri K. M., Dankert M. A. Structural studies of acetan, an exopolysaccharide elaborated by Acetobacter xylinum. Carbohydr Res. 1993 Jul 19;245(2):303–310. doi: 10.1016/0008-6215(93)80079-t. [DOI] [PubMed] [Google Scholar]
  19. Kido N., Torgov V. I., Sugiyama T., Uchiya K., Sugihara H., Komatsu T., Kato N., Jann K. Expression of the O9 polysaccharide of Escherichia coli: sequencing of the E. coli O9 rfb gene cluster, characterization of mannosyl transferases, and evidence for an ATP-binding cassette transport system. J Bacteriol. 1995 Apr;177(8):2178–2187. doi: 10.1128/jb.177.8.2178-2187.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Knauf V. C., Nester E. W. Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid. 1982 Jul;8(1):45–54. doi: 10.1016/0147-619x(82)90040-3. [DOI] [PubMed] [Google Scholar]
  21. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  22. Lee S. J., Romana L. K., Reeves P. R. Sequence and structural analysis of the rfb (O antigen) gene cluster from a group C1 Salmonella enterica strain. J Gen Microbiol. 1992 Sep;138(9):1843–1855. doi: 10.1099/00221287-138-9-1843. [DOI] [PubMed] [Google Scholar]
  23. Leigh J. A., Coplin D. L. Exopolysaccharides in plant-bacterial interactions. Annu Rev Microbiol. 1992;46:307–346. doi: 10.1146/annurev.mi.46.100192.001515. [DOI] [PubMed] [Google Scholar]
  24. Leigh J. A., Walker G. C. Exopolysaccharides of Rhizobium: synthesis, regulation and symbiotic function. Trends Genet. 1994 Feb;10(2):63–67. doi: 10.1016/0168-9525(94)90151-1. [DOI] [PubMed] [Google Scholar]
  25. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  26. Liu D., Haase A. M., Lindqvist L., Lindberg A. A., Reeves P. R. Glycosyl transferases of O-antigen biosynthesis in Salmonella enterica: identification and characterization of transferase genes of groups B, C2, and E1. J Bacteriol. 1993 Jun;175(11):3408–3413. doi: 10.1128/jb.175.11.3408-3413.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Miller G., Feiss M. Cohesive end annealing and the helper-mediated transformation system of phage lambda. Virology. 1981 Mar;109(2):379–390. doi: 10.1016/0042-6822(81)90508-0. [DOI] [PubMed] [Google Scholar]
  28. Paulson J. C., Colley K. J. Glycosyltransferases. Structure, localization, and control of cell type-specific glycosylation. J Biol Chem. 1989 Oct 25;264(30):17615–17618. [PubMed] [Google Scholar]
  29. Pavelka M. S., Jr, Hayes S. F., Silver R. P. Characterization of KpsT, the ATP-binding component of the ABC-transporter involved with the export of capsular polysialic acid in Escherichia coli K1. J Biol Chem. 1994 Aug 5;269(31):20149–20158. [PubMed] [Google Scholar]
  30. Rubens C. E., Heggen L. M., Haft R. F., Wessels M. R. Identification of cpsD, a gene essential for type III capsule expression in group B streptococci. Mol Microbiol. 1993 May;8(5):843–855. doi: 10.1111/j.1365-2958.1993.tb01631.x. [DOI] [PubMed] [Google Scholar]
  31. Saxena I. M., Kudlicka K., Okuda K., Brown R. M., Jr Characterization of genes in the cellulose-synthesizing operon (acs operon) of Acetobacter xylinum: implications for cellulose crystallization. J Bacteriol. 1994 Sep;176(18):5735–5752. doi: 10.1128/jb.176.18.5735-5752.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Semino C. E., Dankert M. A. In vitro biosynthesis of acetan using electroporated Acetobacter xylinum cells as enzyme preparations. J Gen Microbiol. 1993 Nov;139(11):2745–2756. doi: 10.1099/00221287-139-11-2745. [DOI] [PubMed] [Google Scholar]
  33. Stanfield S. W., Ielpi L., O'Brochta D., Helinski D. R., Ditta G. S. The ndvA gene product of Rhizobium meliloti is required for beta-(1----2)glucan production and has homology to the ATP-binding export protein HlyB. J Bacteriol. 1988 Aug;170(8):3523–3530. doi: 10.1128/jb.170.8.3523-3530.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Stankowski J. D., Mueller B. E., Zeller S. G. Location of a second O-acetyl group in xanthan gum by the reductive-cleavage method. Carbohydr Res. 1993 Mar 17;241:321–326. doi: 10.1016/0008-6215(93)80123-v. [DOI] [PubMed] [Google Scholar]
  35. Tolmasky M. E., Crosa J. H. Molecular cloning and expression of genetic determinants for the iron uptake system mediated by the Vibrio anguillarum plasmid pJM1. J Bacteriol. 1984 Dec;160(3):860–866. doi: 10.1128/jb.160.3.860-866.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wong H. C., Fear A. L., Calhoon R. D., Eichinger G. H., Mayer R., Amikam D., Benziman M., Gelfand D. H., Meade J. H., Emerick A. W. Genetic organization of the cellulose synthase operon in Acetobacter xylinum. Proc Natl Acad Sci U S A. 1990 Oct;87(20):8130–8134. doi: 10.1073/pnas.87.20.8130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. de Mendoza D., Rosa A. L. Cloning of mini-mu bacteriophage in cosmids: in vivo packaging into phage lambda heads. Gene. 1985;39(1):55–59. doi: 10.1016/0378-1119(85)90107-6. [DOI] [PubMed] [Google Scholar]

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