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PubMed2001

Effect of cellulose on the digestibility of high starch versus high fat diets in dogs.

The effects of cellulose added in three levels (7, 15 and 20% crude fibre in dry matter) to three different basal diets (a high fat diet, a high starch diet with raw starch and a high starch diet with cooked starch) on apparent digestibility were investigated in eight adult dogs. Cellulose had little effect on the apparent digestibility of fat. In the high fat diet there was no significant decrease, not even at the highest cellulose level (98.3% compared with 98.1% in the basal high fat diet). In the cooked starch diets, fat digestibility decreased from 95.1% in the basal cooked starch diet to 93.8% at the highest cellulose level. In the raw starch diets, digestibility did not decrease with increasing cellulose levels. The apparent digestibility of crude protein was considerably decreased by cellulose in all diets. Starch also decreased protein digestibility and the effects of cellulose and starch appeared to be additive (high fat diet decrease of protein digestibility from 86.7 to 83.5%, cooked starch from 81.6 to 78.6%, raw starch from 79.0 to 70.8%, basal diets to highest cellulose levels, respectively). The apparent digestibility of nitrogen-free extract decreased from 93.9% in the basal cooked starch diet to 84.5% at the highest cellulose level. The figures for the raw starch diets were similar (decrease from 93.4 to 85.9%). Cellulose decreased the apparent digestibility of energy in all diets. This decrease was more marked in the high starch diets (cooked starch decrease from 89.1 to 69.6%, raw starch from 88.9 to 70.2%) than in the high fat diet (decrease from 90.1 to 76.1%). An evaluation of previous data showed that in general fibre has a higher impact on the apparent digestibility of energy in high carbohydrate diets than in low carbohydrate diets. The apparent digestibility of potassium, sodium and chloride was impaired by cellulose. The apparent digestibility of these minerals tended to be lower in the high starch diets, especially in the raw starch diet, and the effect of cellulose was usually more marked in those diets.

Animal Feed↗

PubMed1993

Mode of action of exo- and endo-type cellulases from Irpex lacteus in the hydrolysis of cellulose with different crystallinities.

The mode of action of two highly purified cellulases of exo- and endo-types from Irpex lacteus was investigated by using pure cellulosic substrates with different crystallinities derived from cellulose I. Exo-type cellulase saccharified all celluloses more effectively than endo-type enzyme, and the saccharification activities of both cellulases similarly increased with decreasing crystallinity of cellulose. The DP-lowering activity of exo-type cellulase remained similar for celluloses with higher crystallinity, while this cellulase showed a degradation mode resembling that of the endo-type enzyme for the substrates with lower crystallinity. Compared with exo-type cellulase, endo-type cellulase remarkably decreased the DP of cellulose with higher crystallinity, while this activity was abated for cellulose with lower crystallinity. Thus, the effects of both cellulases became similar in the degradation of amorphous substrates such as H3PO4-treated cellulose. Endo-type cellulase produced several kinds of cellooligosaccharide from all kinds of cellulose used, while the product of the exo-type enzyme was only cellobiose from crystalline cellulose such as cotton and cotton linter even after a 12-h incubation period. The results indicate that each cellulase shows a typical mode of action (exo or endo) for crystalline cellulose, but that their characteristic modes of attack may change with decreasing crystallinity of cellulose.

Cellulase↗

PubMed1992

Response surface analysis of the effects of pH and dilution rate on Ruminococcus flavefaciens FD-1 in cellulose-fed continuous culture.

The ruminal cellulolytic bacterium Ruminococcus flavefaciens FD-1 was grown in cellulose-fed continuous culture with 20 different combinations of pH and dilution rate (D); the combinations were selected according to the physiological pH range of the organism (6.0 to 7.1) and growth rate of the organism on cellulose (0.017 to 0.10 h-1). A response surface analysis was used to characterize the effects of pH and D on the extent of cellulose consumption, growth yield, soluble sugar concentration, and yields of fermentation products. The response surfaces indicate that pH and D coordinately affect cellulose digestion and growth yield in this organism. As expected, the net cellulose consumption increased with increasing D while the fraction of added cellulose that was utilized decreased with increasing D. The effect of changes in pH within the physiological range on cellulose consumption was smaller than that of changes in D. Cellulose degradation was less sensitive to low pH than to high pH. At low Ds (longer retention times), cellulose degradation did not follow first-order kinetics. This decreased rate of cellulose digestion was not due to poor mixing, limitation by other medium components, or preferential utilization of the more amorphous fraction of the cellulose. The cell yield increased from 0.13 to 0.18 mg of cells per mg of cellulose with increasing Ds from 0.02 to 0.06 h-1 and decreased when the pH was shifted from the optimum of 6.5 to 6.8. The effect of pH on cell yield increased with increasing D. The reduced cell yield at low pH appears to be due to both an increase in maintenance energy requirements and a decrease in true growth yield.

Bacteriological Techniques↗

PubMed1997

Competition for cellulose among three predominant ruminal cellulolytic bacteria under substrate-excess and substrate-limited conditions.

Three predominant ruminal cellulolytic bacteria (Fibrobacter succinogenes S85, Ruminococcus flavefaciens FD-1, and Ruminococcus albus 7) were grown in different binary combinations to determine the outcome of competition in either cellulose-excess batch culture or in cellulose-limited continuous culture. Relative populations of each species were estimated by using signature membrane-associated fatty acids and/or 16S rRNA-targeted oligonucleotide probes. Both F. succinogenes and R. flavefaciens coexisted in cellulose-excess batch culture with similar population sizes (58 and 42%, respectively; standard error, 12%). By contrast, under cellulose limitation R. flavefaciens predominated (> 96% of total cell mass) in coculture with F. succinogenes, regardless of whether the two strains were inoculated simultaneously or whether R. flavefaciens was inoculated into an established culture of F. succinogenes. The predominance of R. flavefaciens over F. succinogenes under cellulose limitation is in accord with the former's more rapid adherence to cellulose and its higher affinity for cellodextrin products of cellulose hydrolysis. In batch cocultures of F. succinogenes and R. albus, the populations of the two species were similar. However, under cellulose limitation, F. succinogenes was the predominant strain (approximately 80% of cell mass) in cultures simultaneously coinoculated with R. albus. The results from batch cocultures of R. flavefaciens and R. albus were not consistent within or among trials: some experiments yielded monocultures of R. albus (suggesting production of an inhibitory agent by R. albus), while others contained substantial populations of both species. Under cellulose limitation, R. flavefaciens predominated over R. albus (85 and 15%, respectively), as would be expected by the former's greater adherence to cellulose. The retention of R. albus in the cellulose-limited coculture may result from a combination of its ability to utilize glucose (which is not utilizable by R. flavefaciens), its demonstrated ability to adapt under selective pressure in the chemostat to utilization of lower concentrations of cellobiose, a major product of cellulose hydrolysis, and its possible production of an inhibitory agent.

Animals↗

PubMed1983

Role of bacterial cellulose fibrils in Agrobacterium tumefaciens infection.

During the attachment of Agrobacterium tumefaciens to carrot tissue culture cells, the bacteria synthesize cellulose fibrils. We examined the role of these cellulose fibrils in the attachment process by determining the properties of bacterial mutants unable to synthesize cellulose. Such cellulose-minus bacteria attached to the carrot cell surface, but, in contrast to the parent strain, with which larger clusters of bacteria were seen on the plant cell, cellulose-minus mutant bacteria were attached individually to the plant cell surface. The wild-type bacteria became surrounded by fibrils within 2 h after attachment. No fibrils were seen with the cellulose-minus mutants. Prolonged incubation of wild-type A. tumefaciens with carrot cells resulted in the formation of large aggregates of bacteria, bacterial fibrils, and carrot cells. No such aggregates were formed after the incubation of carrot cells with cellulose-minus A. tumefaciens. The absence of cellulose fibrils also caused an alteration in the kinetics of bacterial attachment to carrot cells. Cellulose synthesis was not required for bacterial virulence; the cellulose-minus mutants were all virulent. However, the ability of the parent bacterial strain to produce tumors was unaffected by washing the inoculation site with water, whereas the ability of the cellulose-minus mutants to form tumors was much reduced by washing the inoculation site with water. Thus, a major role of the cellulose fibrils synthesized by A. tumefaciens appears to be anchoring the bacteria to the host cells, thereby aiding the production of tumors.

Adhesiveness↗

PubMed2002

Microbial cellulose utilization: fundamentals and biotechnology.

Fundamental features of microbial cellulose utilization are examined at successively higher levels of aggregation encompassing the structure and composition of cellulosic biomass, taxonomic diversity, cellulase enzyme systems, molecular biology of cellulase enzymes, physiology of cellulolytic microorganisms, ecological aspects of cellulase-degrading communities, and rate-limiting factors in nature. The methodological basis for studying microbial cellulose utilization is considered relative to quantification of cells and enzymes in the presence of solid substrates as well as apparatus and analysis for cellulose-grown continuous cultures. Quantitative description of cellulose hydrolysis is addressed with respect to adsorption of cellulase enzymes, rates of enzymatic hydrolysis, bioenergetics of microbial cellulose utilization, kinetics of microbial cellulose utilization, and contrasting features compared to soluble substrate kinetics. A biological perspective on processing cellulosic biomass is presented, including features of pretreated substrates and alternative process configurations. Organism development is considered for "consolidated bioprocessing" (CBP), in which the production of cellulolytic enzymes, hydrolysis of biomass, and fermentation of resulting sugars to desired products occur in one step. Two organism development strategies for CBP are examined: (i) improve product yield and tolerance in microorganisms able to utilize cellulose, or (ii) express a heterologous system for cellulose hydrolysis and utilization in microorganisms that exhibit high product yield and tolerance. A concluding discussion identifies unresolved issues pertaining to microbial cellulose utilization, suggests approaches by which such issues might be resolved, and contrasts a microbially oriented cellulose hydrolysis paradigm to the more conventional enzymatically oriented paradigm in both fundamental and applied contexts.

Bacteria↗

PubMed1999

Effects of guar gum and cellulose on cecal enzyme activity and cecal short-chain fatty acids in young and aged mice.

The effects of cellulose or guar gum on cecal enzyme activity and cecal short-chain fatty acids (SCFAs) in young and aged mice were studied. Male Crj:CD-1 (ICR) mice were fed an MF diet for 4 (young mice) or 23 months (aged mice). The MF diet was then replaced with a semisynthetic diet supplemented with 5% guar gum or 5% cellulose. The mice were fed the guar gum or cellulose diet for 3 weeks. There was no significant difference in cecal content between the two diet groups. There were no significant differences in total short-chain fatty acid production between the young mice fed the cellulose and those fed the guar gum diet, and between the aged mice fed the cellulose and guar gum diet. There were significant differences in cecal enzyme activity between the young mice fed the cellulose and those fed the guar gum diet. Beta-glucuronidase activity was significantly higher in the young mice fed the guar gum diet than in those fed the cellulose diet. There were also significant differences in cecal enzyme activity between the aged mice fed the cellulose diet and those fed the guar gum diet. Beta-glucuronidase activity was significantly higher in the aged mice fed the guar gum diet than in these fed the cellulose diet. Beta-glucosidase activity was significantly lower in the aged mice fed the guar gum diet than in those fed the cellulose diet. The effect of cellulose on the microflora between the young and aged mice might be different from the effect of guar gum. The degree of adaptation to the diet of microflora in young and aged mice fed the cellulose diet might differ from that in those fed the guar gum diet. The higher enzyme activities of microflora in aged animals compared to young animals, might have some relation with the incidence of colon cancer in aged animals.

Aging↗

PubMed1998

Effect of ruminal cellulolytic bacterial concentrations on in situ digestion of forage cellulose.

To evaluate the effects of ruminal cellulolytic bacterial concentrations on in vivo cellulose digestion, varying percentages of flaked soybean hulls were substituted for orchardgrass hay in high-forage diets fed to sheep. In two experiments, total and cellulolytic ruminal bacterial concentrations were not affected by diet. No differences were found for in situ digestion of forage cellulose in the first experiment; however, in Exp. 2, ruminal pH and in situ cellulose digestion were lower (P<.01) with a 40% soybean hull diet. In Exp. 3 with four sheep, two diets were compared, one containing 19.6% cellulose from alfalfa meal and the other 64.3% purified wood cellulose. Ruminal pH was lower (P<.02), 9 and 24 h after feeding, for the high-cellulose diet. Total bacterial concentrations did not change with diet; however, the concentration of cellulolytic bacteria increased (P<.05) when the higher cellulose diet was fed. In situ cellulose digestion was not different between diets. In Exp. 4, 3% sodium bicarbonate was added to the high-cellulose diet, and it was fed twice a day. No differences were observed in pH between diets (P>.42). However, the concentration of total ruminal bacteria increased (P<.06), the concentration of cellulolytic bacteria increased (P<.03), and the percentage of cellulolytic bacteria increased (P<.04) when the buffered high-cellulose diet was fed. In situ digestion of alfalfa cellulose at 30 h was not different between diets (P>.60). These data indicate that the concentration of cellulolytic bacteria is not the limiting factor in the digestion of cellulose in the rumen.

Animal Feed↗

PubMed1965

HYDROLYSIS OF FIBROUS COTTON AND REPRECIPITATED CELLULOSE BY CELLULOLYTIC ENZYMES FROM SOIL MICRO-ORGANISMS.

1. The action of cell-free filtrates from Trichoderma koningii was examined on undegraded cellulose in the form of cotton fibres, on degraded cellulose in the form of cellulose powder reprecipitated from phosphoric acid and on the soluble cellulose derivative CM-cellulose. 2. The cell-free filtrates compare favourably with intact cellulolytic micro-organisms in producing complete solubilization of undegraded as well as of degraded types of cellulose. Enzymic solubilization of cotton fibres gives quantitative conversion into glucose. Cellobiase is present. 3. The early enzymic breakdown of cotton fibres is characterized by the formation of very short fibres that increase to a maximum and disappear gradually by conversion into glucose. Disintegration of cotton fibres to short fibres is assisted by shaking, and within 20hr. the enzyme converts a minor fraction (up to 16%) of substrate into soluble products and a major portion (80%) into insoluble short fibres. 4. Maximum enzymic activity on cotton fibres occurs at about pH5.0, measured by the formation of short fibres, and at about pH3.8 on reprecipitated cellulose, measured by solubilization of the substrate. 5. Gluconolactone, glucose and cellobiose fail to produce marked inhibition of the enzymic hydrolysis of cotton fibres to short fibres or of the solubilization of reprecipitated cellulose unless present in amounts comparable with or greater than the initial weight of these two forms of cellulose. 6. The heavy-metal ions Cu(2+), Hg(2+) and Fe(3+) at 2mm concentration give 80-100% decrease in the enzymic breakdown of cotton fibres, measured by the formation of short fibres. At the same concentration Hg(2+) and Fe(3+) but not Cu(2+) also produce 70-100% inhibition of the solubilization of reprecipitated cellulose. 7. The ability to hydrolyse cotton fibres to short fibres and CM-cellulose to sugars is completely lost after heating enzyme preparations for 10min. at 71 degrees .

Cellulose↗

PubMed1997

A randomized clinical trial of oxidized regenerated cellulose adhesion barrier (Interceed, TC7) alone or in combination with heparin.

OBJECTIVE: To compare the efficacy of heparin-saturated oxidized regenerated cellulose absorbable adhesion barrier, Interceed (TC7; Johnson and Johnson Medical Inc., New Brunswick, NJ) to oxidized regenerated cellulose alone for the prevention of postoperative adhesions. DESIGN: Clinical trial. By random assignment, one ovary was wrapped in oxidized regenerated cellulose, and the contralateral ovary was wrapped in oxidized regenerated cellulose saturated with a heparin solution (1,000 U/mL). PATIENT(S): Forty women with defects on both ovaries due to adhesiolysis and/or ovarian cystectomy. MAIN OUTCOME MEASURE: Adhesion formation and raw ovarian surface area were assessed at second-look laparoscopy 10 days to 16 weeks later. RESULT(S): At the second-look laparascopy-adhesions were present on 52.5% (21/40) of the ovaries treated with oxidized regenerated cellulose plus heparin and in 65% (26/40) of the contralateral ovaries treated with oxidized regenerated cellulose alone. For ovaries treated with oxidized regenerated cellulose plus heparin, the raw surface area was reduced from 9.41 +/- 1.27 cm2 (mean +/- SE) at laparotomy to 1.33 +/- 0.52 cm2 at second-look laparoscopy. The corresponding figures for ovaries treated with oxidized regenerated cellulose alone were from 10.24 +/- 1.08 to 1.92 +/- 0.54 cm2, respectively. The mean difference between the reductions in raw surface area (85.9% for oxidized regenerated cellulose plus heparin; 81.3% for oxidized regenerated cellulose alone) was not significantly different from zero (difference = - 0.24 cm2; 95% confidence interval = -2.56 to 3.04). CONCLUSION(S): Adding heparin did not enhance significantly the adhesion-reducing capacity of oxidized regenerated cellulose adhesion barrier when applied to ovarian surfaces after cystectomy and/or ovariolysis at laparotomy. This conclusion is subject to the possibility of a type II error.

Adult↗

PubMed2004

Comparative study of crude and purified cellulose from wheat straw.

A sequential totally chlorine-free procedure for isolation of cellulose from wheat straw was proposed in this study. The dewaxed straw was pretreated with 0.5 M NaOH in 60% methanol at 60 degrees C for 2.5 h under ultrasonic irradiation for 0-35 min and sequentially posttreated with 2% H(2)O(2)-0.2% TAED at pH 11.8 for 12 h at 48 degrees C, which together solubilized 85.3-86.1% of the original hemicelluloses and 91.7-93.2% of the original lignin, respectively. The yield of crude cellulose ranged between 46.2 and 49.2% on a dry weight basis related to wheat straw, which contained 11.2-12.2% residual hemicelluloses and 2.5-2.9% remaining lignin. Further treatment of the corresponding crude cellulosic preparations with 80% acetic acid-70% nitric acid under the condition given yielded 36.8-37.7% of the purified cellulose, which contained minor amounts of bound hemicelluloses (2.5-2.8%) and was relatively free of associated lignin (0.1-0.2%). The isolated crude and purified cellulose samples were comparatively studied by FT-IR and CP/MAS (13)C NMR spectroscopy, and the relative crystallinity was also estimated. The final stage treatment with 80% acetic acid-70% nitric acid decreased the hemicelluloses and lignin associated in the crude cellulose but led to 3.1-5.4% degradation of the original cellulose; in addition, the purity of the obtained cellulose was high. However, it was found that the final stage treatment is not severe enough to cause decrystallization of cellulose. The thermal stability of the purified cellulose is higher than that of the corresponding crude cellulose.

Cellulose↗

PubMed2006

Understanding the biological rationale for the diversity of cellulose-directed carbohydrate-binding modules in prokaryotic enzymes.

Plant cell walls are degraded by glycoside hydrolases that often contain noncatalytic carbohydrate-binding modules (CBMs), which potentiate degradation. There are currently 11 sequence-based cellulose-directed CBM families; however, the biological significance of the structural diversity displayed by these protein modules is uncertain. Here we interrogate the capacity of eight cellulose-binding CBMs to bind to cell walls. These modules target crystalline cellulose (type A) and are located in families 1, 2a, 3a, and 10 (CBM1, CBM2a, CBM3a, and CBM10, respectively); internal regions of amorphous cellulose (type B; CBM4-1, CBM17, CBM28); and the ends of cellulose chains (type C; CBM9-2). Type A CBMs bound particularly effectively to secondary cell walls, although they also recognized primary cell walls. Type A CBM2a and CBM10, derived from the same enzyme, displayed differential binding to cell walls depending upon cell type, tissue, and taxon of origin. Type B CBMs and the type C CBM displayed much weaker binding to cell walls than type A CBMs. CBM17 bound more extensively to cell walls than CBM4-1, even though these type B modules display similar binding to amorphous cellulose in vitro. The thickened primary cell walls of celery collenchyma showed significant binding by some type B modules, indicating that in these walls the cellulose chains do not form highly ordered crystalline structures. Pectate lyase treatment of sections resulted in an increased binding of cellulose-directed CBMs, demonstrating that decloaking cellulose microfibrils of pectic polymers can increase CBM access. The differential recognition of cell walls of diverse origin provides a biological rationale for the diversity of cellulose-directed CBMs that occur in cell wall hydrolases and conversely reveals the variety of cellulose microstructures in primary and secondary cell walls.

Arabidopsis↗

PubMed2001

Prevention of adhesions by surfactants and cellulose derivatives in mice.

OBJECTIVE: To evaluate the efficacy of hydrophobically modified ethyl (hydroxyethyl) cellulose (cellulose), sodium hyaluronate (hyaluronate) and phosphatidylglycerol, in the reduction of adhesion formation. DESIGN: Controlled study. SETTING: Experimental academic unit, Sweden. MATERIAL: NMRI mice. Solutions: (1) cellulose, (2) hyaluronate, (3) phosphatidylglycerol, (4) phosphatidylglycerol and cellulose, and (5) phosphatidylglycerol, cellulose and hyaluronate. INTERVENTIONS: A standard lesion was created in the parietal peritoneum in mice. One of the viscous solutions to be tested, or saline, was given intraperitoneally. MAIN OUTCOME MEASURES: Amount of adhesions found one week postoperatively. RESULTS: Cellulose; phosphatidylglycerol and cellulose; and phosphatidylglycerol, cellulose and hyaluronate all significantly reduced the amount of adhesions (p=0.0002, p=0.002, p < 0.0001), as did the hyaluronate alone (p < 0.05). Phosphatidylglycerol alone did not reduce the amount of adhesions. Combining cellulose with phosphatidylglycerol, or with hyaluronate, did not improve efficacy. CONCLUSION: Cellulose and hyaluronate were effective in reducing the formation of adhesions. Combining cellulose with hyaluronate or phosphatidylglycerol or both did not improve efficacy.

Animals↗

PubMed2002

Caspase-3-dependent pathway mediates apoptosis of human mononuclear cells induced by cellulosic haemodialysis membranes.

BACKGROUND: Mononuclear cells from patients dialysed with cellulosic membranes undergo rapid apoptosis in vitro. The aim of the present study was to determine whether the apoptosis associated with cellulosic haemodialysis membrane shares similar features with the spontaneous apoptosis described in normal monocytes. Thus, we determined whether apoptosis is dependent on caspase-3 activity and is inhibited by lipopolysaccharide (LPS), which are two features of spontaneous apoptosis in normal monocytes. METHODS: We examined mononuclear cells from healthy subjects and from 14 end-stage renal failure patients on haemodialysis with cellulosic membranes (n=7) and non-cellulosic membranes (n=7). Isolated mononuclear cells were cultured for 48 h. To determine the effect of haemodialysis membrane exposure on caspase-3 activity, on mononuclear apoptosis, or both, cells from healthy subjects were cultured in mini-dialysers with the same membrane types that were used in the haemodialysis patients. Caspase-3 active form was determined by flow cytometric analysis using anti-human-active caspase-3 antibodies. The effect of LPS and Ac-DEVD-CHO, a specific inhibitor of active caspase-3, was also evaluated. Cell apoptosis was assessed by the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labelling method. RESULTS: After 48 h of culture, the percentage of mononuclear cells expressing the active form of caspase-3 was greater in patients dialysed with cellulosic membranes than in patients using non-cellulosic membranes and in healthy subjects. This increase in caspase-3 activity was associated with a high rate of apoptosis, which was prevented by Ac-DEVD-CHO, an inhibitor of caspase-3 activity. LPS decreased both apoptosis and caspase-3 activity in mononuclear cells from patients dialysed with cellulosic membranes. Finally, in cells from healthy subjects, both caspase-3 activation and apoptosis were induced after incubation with cellulosic membranes. In contrast, the active form of caspase-3 was not increased in cells cultured with non-cellulosic membranes and was significantly lower than with cellulosic membranes. CONCLUSION: These findings suggest that the apoptosis of mononuclear cells induced by cellulosic haemodialysis membranes occurs through a pathway that is similar to the spontaneous apoptosis of normal monocytes. They additionally suggest that LPS regulates the proteolytic activation of caspase-3.

Adult↗

PubMed2000

Cellulose catabolism by Clostridium cellulolyticum growing in batch culture on defined medium.

A reinvestigation of cellulose degradation by Clostridium cellulolyticum in a bioreactor with pH control of the batch culture and using a defined medium was performed. Depending on cellulose concentration, the carbon flow distribution was affected, showing the high flexibility of the metabolism. With less than 6.7 g of cellulose liter(-1), acetate, ethanol, H(2), and CO(2) were the main end products of the fermentation and cellulose degradation reached more than 85% in 5 days. The electron flow from the glycolysis was balanced by the production of H(2) and ethanol, the latter increasing with increasing initial cellulose concentration. From 6.7 to 29.1 g of cellulose liter(-1), the percentage of cellulose degradation declined; most of the cellulase activity remained on the cellulose fibers, the maximum cell density leveled off, and the carbon flow was reoriented from ethanol to acetate. In addition to that of previously indicated end products, lactate production rose, and, surprisingly enough, pyruvate overflow occurred. Concomitantly the molar growth yield and the energetic yield of the biomass decreased. Growth arrest may be linked to sufficiently high carbon flow, leading to the accumulation of an intracellular inhibitory compound(s), as observed on cellobiose (E. Guedon, M. Desvaux, S. Payot, and H. Petitdemange, Microbiology 145:1831-1838, 1999). These results indicated that bacterial metabolism exhibited on cellobiose was distorted compared to that exhibited on a substrate more closely related to the natural ecosystem of C. cellulolyticum. To overcome growth arrest and to improve degradation at high cellulose concentrations (29.1 g liter(-1)), a reinoculation mode was evaluated. This procedure resulted in an increase in the maximum dry weight of cells (2,175 mg liter(-1)), cellulose solubilization (95%), and end product concentrations compared to a classical batch fermentation with a final dry weight of cells of 580 mg liter(-1) and 45% cellulose degradation within 18 days.

Biodegradation, Environmental↗

PubMed1995

Identification of a second cellulose synthase gene (acsAII) in Acetobacter xylinum.

A second cellulose synthase gene (acsAII) coding for a 175-kDa polypeptide that is similar in size and sequence to the acsAB gene product has been identified in Acetobacter xylinum AY201. Evidence for the presence of this gene was obtained during analysis of A. xylinum mutants in which the acsAB gene was disrupted (I.M. Saxena, K. Kudlicka, K. Okuda, and R.M. Brown, Jr., J. Bacteriol. 176:5735-5752, 1994). Although these mutants produced no detectable cellulose, they exhibited significant cellulose synthase activity in vitro. The acsAII gene was isolated by using an acsAB gene fragment as a probe. The acsAII gene coded for cellulose synthase activity as determined from sequence analysis and study of mutants in which this gene was disrupted. A mutant in which only the acsAII gene was disrupted showed no significant differences in either the in vivo cellulose production or the in vitro cellulose synthase activity compared with wild-type cells. Mutants in which both the acsAII and acsAB genes were disrupted produced no cellulose in vivo and exhibited negligible cellulose synthase activity in vitro, thus confirming that the cellulose synthase activity observed in the acsAB mutants was coded by the acsAII gene. These results establish the presence of an additional gene for cellulose synthase expressed in cells of A. xylinum, yet this gene is not required for cellulose production when cells are grown under laboratory conditions.

Amino Acid Sequence↗

PubMed2006

A CsgD-independent pathway for cellulose production and biofilm formation in Escherichia coli.

Bacterial growth on a surface often involves the production of a polysaccharide-rich extracellular matrix that provides structural support for the formation of biofilm communities. In Salmonella, cellulose is one of the major constituents of the biofilm matrix. Its production is regulated by CsgD and the diguanylate cyclase AdrA that activates cellulose synthesis at a posttranscriptional level. Here, we studied a collection of Escherichia coli isolates, and we found that the ability to produce cellulose is a common trait shared by more than 50% of the tested strains. We investigated the genetic determinants of cellulose production and its role in biofilm formation in the commensal strain E. coli 1094. By contrast with the Salmonella cellulose regulatory cascade, neither CsgD nor AdrA is required in E. coli 1094 to regulate cellulose production. In this strain, an alternative cellulose regulatory pathway is used, which involves the GGDEF domain protein, YedQ. Although AdrA(1094) is functional, it is weakly expressed in E. coli 1094 compared to YedQ, which constitutively activates cellulose production under all tested environmental conditions. The study of cellulose regulation in several other E. coli isolates showed that, besides the CsgD/AdrA regulatory pathway, both CsgD-independent/YedQ-dependent and CsgD-independent/YedQ-independent pathways are found, indicating that alternative cellulose pathways are common in E. coli and possibly in other cellulose-producing Enterobacteriaceae.

Amino Acid Motifs↗

PubMed2006

Manure composition of swine as affected by dietary protein and cellulose concentrations.

An experiment was conducted to investigate the effects of reducing dietary CP and increasing dietary cellulose concentrations on manure DM, C, N, S, VFA, indole, and phenol concentrations. Twenty-two pigs (105 kg initial BW) were fed diets containing either 14.5 or 12.0% CP, in combination with either 2.5 or 8.7% cellulose. Pigs were fed twice daily over the 56-d study, with feed intake averaging 2.74 kg/d. Feces and urine were collected after each feeding and added to the manure storage containers. Manure storage containers were designed to provide a similar unit area per animal as found in industry (7,393 cm2). Before sampling on d 56, the manure was gently stirred to obtain a representative sample for subsequent analyses. An interaction of dietary CP and cellulose was observed for manure acetic acid concentration, in that decreasing CP lowered acetic acid in pigs fed standard levels of cellulose but increased acetic acid in pigs fed greater levels of cellulose (P = 0.03). No other interactions were noted. Decreasing dietary CP reduced manure pH (P = 0.01), NH4 (P = 0.01), isovaleric acid (P = 0.06), phenol (P = 0.05), and 4-ethyl phenol (P = 0.02) concentrations. Increasing dietary cellulose decreased pH (P = 0.01) and NH4 (P = 0.07) concentration but increased manure C (P = 0.03), propionic acid (P = 0.01), butyric acid (P = 0.03), and cresol (P = 0.09) concentrations in the manure. Increasing dietary cellulose also increased manure DM (P = 0.11), N (P = 0.11), and C (P = 0.02) contents as a percentage of nutrient intake. Neither cellulose nor CP level of the diet affected manure S composition or output as a percentage of S intake. Headspace N2O concentration was increased by decreasing dietary CP (P = 0.03) or by increasing dietary cellulose (P = 0.05). Neither dietary CP nor cellulose affected headspace concentration of CH4. This study demonstrates that diets differing in CP and cellulose content can significantly impact manure composition and concentrations of VFA, phenol, and indole, and headspace concentrations of N(2)O, which may thereby affect the environmental impact of livestock production on soil, air, and water.

Animal Feed↗