Cloning of cellulose synthesis related genes from Acetobacter xylinum ATCC23769 and ATCC53582: comparison of cellulose synthetic ability between strains.

About 14.5 kb of DNA fragments from Acetobacter xylinum ATCC23769 and ATCC53582 were cloned, and their nucleotide sequences were determined. The sequenced DNA regions contained endo-beta-1,4-glucanase, cellulose complementing protein, cellulose synthase subunit AB, C, D and beta-glucosidase genes. The results from a homology search of deduced amino acid sequences between A. xylinum ATCC23769 and ATCC53582 showed that they were highly similar. However, the amount of cellulose production by ATCC53582 was 5 times larger than that of ATCC23769 during a 7-day incubation. In A. xylinum ATCC53582, synthesis of cellulose continued after glucose was consumed, suggesting that a metabolite of glucose, or a component of the medium other than glucose, may be a substrate of cellulose. On the other hand, cell growth of ATCC23769 was twice that of ATCC53582. Glucose is the energy source in A. xylinum as well as the substrate of cellulose synthesis, and the metabolic pathway of glucose in both strains may be different. These results suggest that the synthesis of cellulose and the growth of bacterial cells are contradictory.

[1]  D. Danon,et al.  SYNTHESIS OF CELLULOSE BY ACETOBACTER XYLINUM V. Ultrastructure of Polymer , 1962 .

[2]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[3]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[4]  N. Tonouchi,et al.  A beta-glucosidase gene downstream of the cellulose synthase operon in cellulose-producing Acetobacter. , 1997, Bioscience, biotechnology, and biochemistry.

[5]  R. Brown,et al.  Cloning and sequencing of the cellulose synthase catalytic subunit gene of Acetobacter xylinum , 1990, Plant Molecular Biology.

[6]  R. Brown,et al.  Identification of a second cellulose synthase gene (acsAII) in Acetobacter xylinum , 1995, Journal of bacteriology.

[7]  K. Tajima,et al.  Structural analyses of new tri- and tetrasaccharides produced from disaccharides by transglycosylation of purified Trichoderma viride β-glucosidase , 1999, Glycoconjugate Journal.

[8]  M. Van Montagu,et al.  Rapid mapping of transposon insertion and deletion mutations in the large Ti-plasmids of Agrobacterium tumefaciens. , 1979, Nucleic acids research.

[9]  K. Tajima,et al.  Cloning and sequencing of the beta-glucosidase gene from Acetobacter xylinum ATCC 23769. , 2001, DNA research : an international journal for rapid publication of reports on genes and genomes.

[10]  S. Horinouchi,et al.  Addition of a Small Amount of an Endoglucanase Enhances Cellulose Production by A cetobacter xylinum , 1995 .

[11]  A. Szalay,et al.  Site-directed Tn5 and transplacement mutagenesis: methods to identify symbiotic nitrogen fixation genes in slow-growing Rhizobium. , 1987, Methods in enzymology.

[12]  B. Henrissat,et al.  Multidomain architecture of beta-glycosyl transferases: implications for mechanism of action , 1995, Journal of bacteriology.

[13]  K. Tajima,et al.  Effects of endogenous endo-beta-1,4-glucanase on cellulose biosynthesis in Acetobacter xylinum ATCC23769. , 2002, Journal of bioscience and bioengineering.

[14]  J. Blatny,et al.  A new gene required for cellulose production and a gene encoding cellulolytic activity in Acetobacter xylinum are colocalized with the bcs operon , 1994, Journal of bacteriology.

[15]  Y. Pyun,et al.  Evidence that a beta-1,4-endoglucanase secreted by Acetobacter xylinum plays an essential role for the formation of cellulose fiber. , 1998, Bioscience, biotechnology, and biochemistry.

[16]  C. Wang,et al.  The dissimilation of glucose and gluconate by Acetobacter xylinum. 1. The origin and the fate of triose phosphate. , 1964, The Biochemical journal.

[17]  J. H. Boom,et al.  Regulation of cellulose synthesis in Acetobacter xylinum by cyclic diguanylic acid , 1987, Nature.