Cellulose synthesis by acetobacter xylinum II. Investigation into the relation between cellulose synthesis and cell envelope components

1975 ◽  
Vol 381 (1) ◽  
pp. 97-108 ◽  
Author(s):  
David Cooper ◽  
R.St.John Manley
1963 ◽  
Vol 41 (1) ◽  
pp. 1691-1702 ◽  
Author(s):  
T. E. Webb ◽  
J. Ross Colvin

The production of cellulose by lysozyme lysates of Acetobacter xylinum is similar to that of a suspension of whole cells, in contrast to the negative results obtained with previous "cell-free" preparations. The results of differential centrifugation of these lysates suggests that most of the enzymes required for cellulose synthesis from glucose normally are held by the cell envelope and are not located in the cytoplasm. However, a heat-stable cofactor(s) is present in the supernatant derived from the cell contents which may stimulate cellulose synthesis by the cell envelopes.The addition of extracts from a number of plant sources increased cellulose synthesis by whole cells of A. xylinum. In particular, the supernatant prepared by centrifugation of an homogenate of tomatoes increased bacterial cellulose production at pH 6 by a factor of 3. Both dialyzable and non-dialyzable substances in the extract are responsible. Fractionation of the non-dialyzable portion of the extract by column chromatography suggests that the overall increase is due to additive effects of several compounds. Here also the compounds appear to act upon the bacterial cell envelope.


1977 ◽  
Vol 23 (6) ◽  
pp. 701-709 ◽  
Author(s):  
J. Ross Colvin ◽  
Gary G. Leppard

The morphological aspects of biosynthesis of cellulose by Acetobacter xylinum and Acetobacter acetigenus were studied by transmission electron microscopy of both freeze-etch replicas and sections of cellulose-free cells in suspension culture before and subsequent to the induction of cellulose synthesis. Also examined were freshly synthesized, thoroughly washed, cellulose pellicles. Thin sections of rapidly dividing, glucose-metabolizing cells of both species showed irregular features on the cell surface including a small polar invagination which sometimes contained or was associated with fibrils as fine as 3 nm in diameter of a substance which stains with electron-microscopic counterstains. Cellulose microfibrils in thin sections of freshly synthesized pellicles were coated with a surface material which also stained with the same counterstains (uranyl ions and lead ions). The effect of air-drying on freshly synthesized cellulose was striking. When examined by freeze-etching, thoroughly washed, never air-dried pellicles of both species showed a nascent form of cellulose fibril which consisted of a central, dense core surrounded by a sheath of amorphous gel. This sheath may be up to 100 nm wide. When the pellicle was air-dried and rehydrated before freeze-etching, the amorphous sheath was rare and shrunken but ordinary microfibrils of classical dimensions were visible. The sheath and core are sometimes closely associated with the envelope of the cells of both species. These observations can be interpreted in the context of recent advances in cellulose synthesis by assuming that chains of an initial, highly hydrated, intermediate polyglucan are released from the cell and that such chains associate to form a nascent fibril external to the cell but associated with the cell envelope. Air-drying of nascent fibrils converts them to classical microfibrils and this conversion is considered here in molecular terms.


1963 ◽  
Vol 41 (8) ◽  
pp. 1691-1702 ◽  
Author(s):  
T. E. Webb ◽  
J. Ross Colvin

The production of cellulose by lysozyme lysates of Acetobacter xylinum is similar to that of a suspension of whole cells, in contrast to the negative results obtained with previous "cell-free" preparations. The results of differential centrifugation of these lysates suggests that most of the enzymes required for cellulose synthesis from glucose normally are held by the cell envelope and are not located in the cytoplasm. However, a heat-stable cofactor(s) is present in the supernatant derived from the cell contents which may stimulate cellulose synthesis by the cell envelopes.The addition of extracts from a number of plant sources increased cellulose synthesis by whole cells of A. xylinum. In particular, the supernatant prepared by centrifugation of an homogenate of tomatoes increased bacterial cellulose production at pH 6 by a factor of 3. Both dialyzable and non-dialyzable substances in the extract are responsible. Fractionation of the non-dialyzable portion of the extract by column chromatography suggests that the overall increase is due to additive effects of several compounds. Here also the compounds appear to act upon the bacterial cell envelope.


1961 ◽  
Vol 39 (3) ◽  
pp. 493-497 ◽  
Author(s):  
J. Ross Colvin ◽  
S. M. Martin ◽  
G. G. Dearing

Extracellular glucose-1-phosphate (G-1-P) was identified and isolated from suspensions of intact, washed, glucose-metabolizing cells of Acetobacter xylinum. The presence of this extracellular hexose phosphate was closely correlated with active glucose metabolism of the cells but the compound was not the immediate precursor of bacterial cellulose. The extracellular G-1-P was not a degradation product of uridine diphosphate glucose but could have been a fragment of a still larger, more complex molecule. Possible modes of transport of the ester across the bacterial cell membrane are discussed.


2005 ◽  
Vol 52 (3) ◽  
pp. 691-698 ◽  
Author(s):  
Alina Krystynowicz ◽  
Maria Koziołkiewicz ◽  
Agnieszka Wiktorowska-Jezierska ◽  
Stanisław Bielecki ◽  
Emilia Klemenska ◽  
...  

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 unknown. 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.


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