Characterization of exoglucanase and synergistic hydrolysis of cellulose in Clostridium stercorarium

Abstract A cellobiohydrolase component was isolated from an anaerobic thermophilic cellulolytic bacterium, Clostridium stercorarium . When acting alone, the enzyme showed minimal activity towards ordered substrates such as cellulose and filter paper but it has been shown to attack phosphoric-acid swollen cellulose giving cellobiose as principal product. When recombined with endoglucanase it did allow an extensive hydrolysis demonstrating a marked synergism in the action of those two components; the addition of β-glucosidase resulted in a further increase in activity.     

Variations of soil structure and microbial population in a compost amended soil

Changes in soil structure and in microbial population were recorded in a long term field experiment over the growing season of maize (June–November). Determinations were made on samples from plots which had received, for two years, the following treatments: mineral fertilizers, farmyard manure and three rates of compost. Seasonal variations were observed for the stability of soil aggregates, total porosity, pore size distribution, mycorrhizal infection and aerobic cellulolytic microorganisms. The stability of the soil aggregates changed in a similar way to that found for both mycorrhizal infection and the number of aerobic cellulolytic microorganisms. Physical characteristics were not affected in any instance by the organic dressings and microbiological populations were generally influenced only by the higher doses of compost.     

Biochemical characteristics of cellulose and a green alga degradation by Gilvimarinus japonicas 12-2T,and its application potential for seaweed saccharification

ABSTRACT

Cellulose is one of the major constituents of seaweeds, but reports of mechanisms in microbial seaweed degradation in marine environment are limited, in contrast to the multitude of reports for lignocellulose degradation in terrestrial environment. We studied the biochemical characteristics for marine cellulolytic bacterium Gilvimarinus japonicas 12-2^T in seaweed degradation. The bacterial strain was found to degrade green and red algae, but not brown algae. It was shown that the bacterial strain employs various polysaccharide hydrolases (endocellulase, agarase, carrageenanase, xylanase, and laminarinase) to degrade seaweed polysaccharides. Electrophoretic analysis and peptide sequencing showed that the major protein bands on the electrophoresis gel were homologous to known glucanases and glycoside hydrolases. A seaweed hydrolysate harvested from the bacterial culture was found useful as a substrate for yeasts to produce ethanol. These findings will provide insights into possible seaweed decomposition mechanisms of Gilvimarinus, and its biotechnological potential for ethanol production from inedible seaweeds.     

Effect of sulfur supplements on cellulolytic rumen micro-organisms and microbial protein synthesis in cattle fed a high fibre diet

AIM: To examine the effect of sulfur-containing compounds on the growth of anaerobic rumen fungi and the fibrolytic rumen bacteria Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogenes in pure culture and within the cattle rumen. METHODS AND RESULTS: The effect of two reduced sulfur compounds, 3-mercaptopropionic acid (MPA) or 3-mercapto-1-propanesulfonic acid as the sole S source on growth of pure fibroyltic fungal and bacterial cultures showed that these compounds were capable of sustaining growth. An in vivo trial was then conducted to determine the effect of sulfur supplements (MPA and sodium sulfate) on microbial population dynamics in cattle fed the roughage Dichanthium aristatum. Real-time PCR showed significant increases in fibrolytic bacterial and fungal populations when cattle were supplemented with these compounds. Sulfate supplementation leads to an increase in dry matter intake without a change in whole tract dry matter digestibility. CONCLUSIONS: Supplementation of low S-containing diets with either sodium sulfate or MPA stimulates microbial growth with an increase in rumen microbial protein supply to the animal. SIGNIFICANCE AND IMPACT OF THE STUDY: Through the use of real-time PCR monitoring, a better understanding of the effect of S supplementation on discrete microbial populations within the rumen is provided.     

Quantification by real-time PCR of cellulolytic bacteria in the rumen of sheep after supplementation of a forage diet with readily fermentable carbohydrates: effect of a yeast additive

AIM: To examine the effect of concentrate and yeast additive on the number of cellulolytic bacteria in the rumen of sheep. METHODS AND RESULTS: Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens were quantified using real-time PCR (targeting 16S rDNA) in parallel to cellulolytic flora enumeration with cultural techniques. Whatever the conditions tested, R. flavefaciens was slightly more abundant than F. succinogenes, with both species outnumbering R. albus. Before feeding, the shift from hay to hay plus concentrate diet had no effect on rumen pH and on the number of the three specie; while after feeding, the concentrate-supplemented diet induced a decrease (-1 log) of the number of the three species concomitant with the rumen acidification. Overall, the presence of the live yeast resulted in a significant increase (two- to fourfold) of the Ruminococci. CONCLUSION: The use of real-time PCR allowed us to show changes in the number of cellulolytic bacterial species in vivo in response to diet shift and additives that could not be as easily evidenced by classical microbial methods. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributes to the understanding of the negative impact of readily fermentable carbohydrates on rumen cellulolysis and the beneficial effect of yeast on rumen fermentation.     

Biochanin A improves fibre fermentation by cellulolytic bacteria

Aims

The objective was to determine the effect of the isoflavone biochanin A (BCA) on rumen cellulolytic bacteria and consequent fermentative activity.

Methods and Results

When bovine microbial rumen cell suspensions (n = 3) were incubated (24 h, 39°C) with ground hay, cellulolytic bacteria proliferated, short‐chain fatty acids were produced and pH declined. BCA (30 μg ml^?1) had no effect on the number of cellulolytic bacteria or pH, but increased acetate, propionate and total SCFA production. Addition of BCA improved total digestibility when cell suspensions (n = 3) were incubated (48 h, 39°C) with ground hay, Avicel, or filter paper. Fibrobacter succinogenes S85, Ruminococcus flavefaciens 8 and Ruminococcus albus 8 were directly inhibited by BCA. Synergistic antimicrobial activity was observed with BCA and heat killed cultures of cellulolytic bacteria, but the effects were species dependent.

Conclusions

These results indicate that BCA improves fibre degradation by influencing cellulolytic bacteria competition and guild composition.

Significance and Impact of the Study

BCA could serve as a feed additive to improve cellulosis when cattle are consuming high‐fibre diets. Future research is needed to evaluate the effect of BCA on fibre degradation and utilization in vivo.     

Soil microbial communities in the zone of uranium deposits of the Central Aldan (South Yakutia)

Specific features of soil microbial colonies in industry-affected mountainous-taiga permafrost landscapes in the El’kon uranium ore district on the territory of South Yakutia are revealed: a high number of ecological trophic groups of microorganisms (2.0 × 10³ ?7.6 × 10⁷ cells/g), comparable to the density of microbes in meadow steppe soils of Central Yakutia and a special nature of their distribution over the soil profile depending on the uranium content of the soil. In the soil of a uranium-contaminated pit, the number of individuals in all examined groups of microorganisms increases with a decrease in uranium content to 161 ppm. In the remaining samples of this pit, the disappearance of microorganisms or their decline by one or two orders of magnitude with increasing uranium content of the soil is observed. A different situation is with microorganisms in the soil of native landscape: Their numbers remain there high over the entire soil profile. Estimated correlations between the number of individuals of the main ecological trophic groups of microorganisms and factors such as uranium content, temperature, and humidity of soil shows that the relationship with the concentration of this radionuclide was strongly negative (r = ?0.6) in the radiation-contaminated alluvial soil, while a strict relationship between the number of microorganisms and soil temperature was revealed for the uncontaminated soil (r = ±1).     

Cellulolytic streptomycetes from <Emphasis Type="Italic">Sphagnum</Emphasis> peat bogs and factors controlling their activity

Two strains of Actinobacteria, ACTY and ACTR, were isolated from cellulolytic microbial communities obtained from an ombrotrophic Sphagnum peat bog. The strains were able to degrade cellulose, the main component of plant phytomass in this ecosystem. On the basis of their phenotypic and phylogenetic characteristics, the strains were identified as members of the genus Streptomyces. The isolates developed on media without available nitrogen sources and hydrolyzed cellulose within a temperature range of 5–25°C and in the pH interval from 4.5 to 6.0; they also exhibited acetylene reduction activity. Comparative analysis of the rates of cellulose degradation by the peat-inhabiting streptomyces at 5, 15, and 25°C and at pH values of 4.5 and 6.0, with and without a source of available nitrogen in the medium, indicated that high acidity and low temperatures, typical for boreal Sphagnum peat bogs, are the main factors limiting the growth and hydrolytic activity of these bacteria.     

混合培养对固氮菌和纤维素分解菌生长及固氮的影响

对筛选的自生固氮菌和纤维素分解菌进行混合培养 ,研究了菌数与菌液含氮量的变化情况 ,并与其单独培养进行了比较。实验证明 :在混合培养条件下 ,两种菌能相互利用、相互促进 ,混合培养液的菌数增加 ,固氮菌的固氮能力提高。这两种菌可混合培养制成混合菌剂。。