Preferences help
enabled [disable] Abstract
Number of results
2005 | 52 | 3 | 691-698
Article title

Molecular basis of cellulose biosynthesis disappearance in submerged culture of Acetobacter xylinum

Title variants
Languages of publication
Acetobacter xylinum strains are known as very efficient producers of bacterial cellulose which, due to its unique properties, has great application potential. One of the most important problems faced during cellulose synthesis by these bacteria is generation of cellulose non-producing cells, which can appear under submerged culture conditions. The reasons of this remain unknow. These studies have been undertaken to compare at the molecular level wild-type, cellulose producing (Cel+) A. xylinum strains with Cel- forms of cellulose-negative phenotype. Comparison of protein profiles of both forms of A. xylinum by 2D electrophoresis allowed for the isolation of proteins which were produced exclusively by either Cel+ or Cel- cells. Sequences of peptides derived from these proteins were aligned with those of proteins deposited in databases. This analysis revealed that Cel- cells lacked two enzymes: phosphoglucomutase and glucose-1-phosphate uridylyltransferase, which generates UDP-glucose being the substrate for cellulose synthase. DNA was analyzed by ligation-mediated PCR carried out at low denaturation temperature (PCR-MP). Two DNA fragments of different thermal stability (218 and 217 bp) were obtained from the DNA of Cel+ and Cel- forms, respectively. The only difference between these Cel- and Cel+ DNA fragments is deletion of one T residue. Alignment of those two sequences with those deposited in the GenBank database revealed that similar fragments are present in the genomes of some bacterial cellulose producers and are located downstream from open reading frames (ORF) encoding phosphoglucomutase. The meaning of this observation is discussed.
Physical description
  • Institute of Technical Biochemistry, Technical University of Lodz, Łódź, Poland
  • Institute of Technical Biochemistry, Technical University of Lodz, Łódź, Poland
  • Institute of Technical Biochemistry, Technical University of Lodz, Łódź, Poland
  • Institute of Technical Biochemistry, Technical University of Lodz, Łódź, Poland
  • Institute of Biotechnology and Antibiotics, Warszawa, Poland
  • Institute of Biotechnology and Antibiotics, Warszawa, Poland
  • Institute of Biotechnology and Antibiotics, Warszawa, Poland
  • Bielecki S, Krystynowicz A, Turkiewicz M, Kalinowska H (2005) Bacterial cellulose. In Biotechnology of Biopolymers, Steinbuchel A, Doi Y, eds, vol. 1, pp 381-434. Willey-VCH, Weinheim.
  • Bartlett DH, Silverman M (1989) Nucleotide sequences of IS492, a novel insertion sequences causing variation in extracellular polysaccharide production in the marine bacterium Pseudomonas atlantica. J Bacteriol 171: 1763-1766.
  • Brautaset T, Standal R, Fjaervik E, Valla S (1994) Nucleotide sequence and expression analysis of the Acetobacter xylinum phosphoglucomutase gene. Microbiology 140: 1183-1188.
  • Coucheron DH (1991) An Acetobacter xylinum insertion sequence element associated with inactivation of cellulose production. J Bacteriol 173: 5723-5731.
  • Dybczyński I, Płucienniczak A (1988) A protocol for DNA fragment extraction from polyacrylamide gels. Biotechniques 6: 924-926.
  • Easson DD Jr, Sinskey AJ, Peoples OP (1987) Isolation of Zoogloea ramigera I-16 M exopolysaccharide biosynthetic genes and evidence for instability within this region. J Bacteriol 169: 4518-4524.
  • Edwards KJ, Jay AJ, Colquhoun IJ, Morris VJ, Gasson MJ, Griffin AM (1999) Generation of a novel polysaccharide by inactivation of the aceP gene from the acetan biosynthetic pathway in Acetobacter xylinum. Microbiology 145: 1499-1506.
  • Hotte B, Roth-Arnold I, Puhler A, Simon R (1990) Cloning and analysis of a 35.3 kb DNA region involved in exopolysaccharide production by Xanthomonas campestris. J Bacteriol 172: 2804-2807.
  • Krystynowicz A, Czaja W, Wiktorowska-Jezierska A, Goncalves-Miśkiewicz M, Turkiewicz M, Bielecki S (2002) Factors affecting the yield and properties of bacterial cellulose. J Ind Microbiol Biotechnol 29: 189-195.
  • Leisinger T, Wiemken A, Ettlinger L (1966) Űber cellulosefreie Mutanten von Acetobacter xylinus. Arch Microbiol 54: 21-36.
  • Maniatis T, Sambrook J, Fritsch EF (1989) Molecular Cloning. A Laboratory Manual. 2 edn.
  • Masny A, Płucienniczak A (2003) Ligation mediated PCR performed at low denaturation temperatures - PCR melting profiles. Nucleic Acids Res 31: e114.
  • Nakai T, Moriya A, Tonouchi N, Tsuchida T, Yoshinaga F, Horinouchi S, Sone Y, Mori H, Sakai F, Hayashi T (1998) Control of expression by the cellulose synthase (bcsA) promoter region from Acetobacter xylinum BPR 2001. Gene 213: 93-100.
  • Protein Electrophoresis, Technical manual (1999) Amersham Pharmacia Biotech.
  • Prust C, Hoffmeister M, Liesegang H, Wiezer A, Fricke WF, Ehrenreich A, Gottschalk G, Deppenmeier U (2005) Complete genome sequence of the acetic acid bacterium Gluconobacter oxydans. Nat Biotechnol 23: 195-200.
  • Ring DF, Nashed W, Dow T (1986) Liquid loaded pad for medical applications. US Patent 45884000.
  • Ross P, Mayer R, Benziman M (1991) Cellulose biosynthesis and function in bacteria. Microbiol Rev 55: 35-58.
  • Schramm M, Hestrin S (1954) Factors affecting production of cellulose at the air/liquid interface of a culture of Acetobacter xylinum. J Gen Microbiol 11: 123-129.
  • Son HJ, Heo MS, Kim YG, Lee SJ (2001) Optimization of fermentation conditions for the production of bacterial cellulose by a newly isolated Acetobacter sp. A9 in shaking cultures. Biotechnol Appl Biochem 33: 1-5.
  • Tajima K, Nakajima K, Yamashita H, Shiba T, Munekata M, Takai M (2001) Cloning and sequencing of the beta-glucosidase gene from Acetobacter xylinum ATCC 23769. DNA Res 8: 263-269.
  • Tamaki T, Horinouchi S, Fukaya M, Okumura H, Kawamura Y, Beppu T (1989) Nucleotide sequence of the membrane-bound aldehyde dehydrogenase gene from Acetobacter polyoxogenes. J Biochem (Tokyo) 106: 541-544.
  • Thurner C, Vela C, Thony-Meyer L, Meile L, Teuber M (1997) Biochemical and genetic characterization of the acetaldehyde dehydrogenase complex from Acetobacter europaeus. Arch Microbiol 168: 81-91.
  • Turner RJ, Lu Y, Switzer RL (1994) Regulation of the Bacillus subtilis pyrimidine biosynthetic gene cluster by an autogenous transcriptional attenuation mechanism. J Bacteriol 176: 3708-3722.
  • Westemeier R, Naven T (2002) Proteomics in Practise. A Laboratory Manual of Proteome Analysis. Wiley-VCH.
  • Wong HC, Fear AL, Calhoon RD, Eichinger GH, Mayer R, Amikam D, Benziman M, Gelfand DH, Meade JH, Emerick AW, Bruner R, Ben-Bassat A, Tal R (1990) Genetic organization of the cellulose synthase operon in Acetobacter xylinum. Proc Natl Acad Sci USA 87: 8130-8134.
  • Valla S, Coucheron DH, Kjosbakken J (1987) The plasmids of Acetobacter xylinum and their interaction with the host chromosome. Mol Gen Genet 208: 76-83.
  • Valla S, Coucheron DH, Fjaervik E, Kjosbakken J, Weinhouse H, Ross P, Amikam D, Benziman M (1989) Cloning of a gene involved in cellulose biosynthesis in Acetobacter xylinum: complementation of cellulose-negative mutant by the UDPG pyrophosphorylase structural gene. Mol Gen Genet 217: 26-30.
  • Yamada Y (2000) Transfer of Acetobacter oboediens Sokollek et al. 1998 and Acetobacter intermedius Boesch et al., 1998 to the genus Gluconacetobacter as Gluconacetobacter oboediens comb. nov. and Gluconacetobacter intermedius comb. nov. Int J Syst Evol Microbiol 50: 2225-2227.
  • Yanofsky C (2000) Transcription attenuation: once viewed as a novel regulatory strategy. J Bacteriol 182: 1-8.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.