Unraveling aerobic cultivable cellulolytic microorganisms within the gastrointestinal tract of sheep (Ovis aries) and their evaluation for cellulose biodegradation

Anaerobic cellulolytic microbes in gastrointestinal tract (GT) of ruminants have been well-documented, however, knowledge of aerobic microbes with cellulolytic activities in ruminant GT is comparably limited. Here, we unraveled aerobic cultivable cellulolytic microbes in GT of Ujimqin sheep (Ovis aries) and evaluated the cellulolytic potential of promising isolates. Twenty-two strains were found to possess cellulose degrading potential by Congo-red staining and phylogenetic analysis of the 16S rDNA/ITS sequence revealed that all strains belonged to nine genera, i.e., Bacillus, Streptomyces, Pseudomonas, Lactobacillus, Brachybacterium, Sanguibacter, Rhizobium, Fusarium, and Aspergillus. Strains with high cellulolytic activities were selected to further evaluate the various enzyme activities on lignocellulosic alfalfa hay (Medicago sativa). Among them, isolate Bacillus subtilis RE2510 showed the highest potential of cellulose degradation considering the high endoglucanase (0.1478 ± 0.0014 IU ml-1), exoglucanase (0.1735 ± 0.0012 IU ml-1) and β-glucosidase (0.3817 ± 0.0031 IU ml-1) after 10-day incubation with alfalfa hay. A significant destruction effect of the cellulose structure and the attachment of B. subtilis RE2510 to the hay were also revealed by using scanning electron microscope. This study expands our knowledge of aerobic cellulolytic isolates from GT of sheep and also highlights their potential application as microbial additive in the aerobic process of cellulose bioconversion.

Exploring cellulolytic microorganisms from coffee industry by-products and their enzyme properties

Cellulolytic microorganism has immense potential due to their cellulase production, enzyme complexity and widespread habitat of life. This study was conducted to obtain microbial cellulase with vast industrial applicability from the coffee industry by-product in East Java, Indonesia. Fifty-four isolates with significant clear zone formation were obtained by Congo red staining in CMC agar plates. Eighteen bacteria, two yeasts and two moulds with high cellulolytic index were subjected to protein content determination as well as reducing sugar analysis in various conditions such as pH, temperature, addition of metal ions, surfactant and inhibitor agent. The specific activity measurements of all the crude enzymes result in the highest value of cellulase activity produced by isolate C12 which was 0.401 ± 0.018 U/mg. This enzyme activity was known to be optimum at 50°C and pH 9. It was also stimulated by K+, Na+, Mg2+, Fe3+, and SDS. However, the enzyme activity was inhibited by EDTA at 10 mM concentration. The use of coffee industry by-products as the source of cellulolytic microorganisms offers a promising approach for its various types of indigenous microorganisms and their unique property of cellulase produced that is useful for industrial application.

QUALITY ANALYSIS OF "KOSMOS" TKKS COMPOSE USING SELULOTIC MICROORGANISM

This study aimed to determine the comparison of the chemical composition of oil oil palm empty friut bunches (TKKS) compost using cellulolytic microorganisms and TKKS compost. This research is a descriptive study that describes the chemical composition of compost with cellulolytic microorganisms. The treatments used in this study were the differences in the ratio of TKKS and cow dung KM1 (100: 0), KM2 (90:10), KM3 (80:20), KM4 (70:30), KM5 (60:40) and KM6 ( 50:50). This study's results were the selected treatment, namely KM5 with an N content of 0.73%, C-Organic 9.28%, C / N ratio 12.29%, P 0.42%, K 3.99%, while the TKKS content only. N 6.28%, C-Organic 14.19%, C / N ratio 2.26%, P 1.88%, K 2.51% and Mg 1.61%.

Endoglucanase Activity of Novel Aspergillus sydowii Isolate WIU-01 and Application for Hydrophyte Degradation

<p>Many eutrophic lakes contain rapidly growing hydrophytes. Overgrown biomass is usually me-chanically harvested and thrown away, leading to resource waste and secondary environmental pollution. Microbial degradation is an economically and environmentally friendly approach for managing hydrophytic waste, fuelling the search for efficient biomass degraders. Here, we present isolation and characterization of Aspergillus sydowii WIU-01, a novel cellulolytic fungus. Strain WIU-01 was isolated from air. The degradation rate (29.75 vs. 21.95%) and endoglucanase activi-ty (0.31 vs. 0.16 U mL–1) of the fungus were higher in Canna indica (emergent plant) medium than in Hydrilla verticillata (submerged plant) medium, accordingly. Further, fungal endoglucanase ac-tivity was significantly positively correlated with the degradation rate, neutral detergent fiber con-tent, and acid detergent fiber content of hydrophyte powder. Fungal biomass was significantly negatively correlated with reducing sugar and cellulose content of hydrophyte medium, but was significantly positively correlated with hemicellulose, acid detergent lignin, and ash content of the medium. Collectively, these observations indicate that A. sydowii decomposes emergent and sub-merged plant mass without acid–base sample pretreatment, albeit its endoglucanase activity is rela-tively low. This highlights the role of cellulolytic microorganisms in the natural environment and the notion that the environment can be a source of cellulolytic microorganisms for potential envi-ronmentally friendly applications.</p>

Isolation and Characterization of Cellulase Producing Bacteria for the Purpose of Converting Spent Mushroom Edible Canna-Waste into Organic Fertilizer

Converting spent mushroom substrates into organic fertilizer helps to tackle the problem of pollution in edible canna starch processing villages and adds new value to the production chain of edible canna. To successfully turn the spent substrates into compost, there is certainly an indispensable role for cellulolytic microorganisms, in which Bacillus strains are always important. Several bacterial strains have been isolated from spent edible canna substrate after cultivation of monkey head mushroom in this study. Among isolated strains, the strain NDK5 has been selected exhibiting the highest cellulolytic activities with solubilization indexes of 6.14 and 18.3 mm for the ratio between the halo zone diameters and the colony diameters in the point cultivation method (SIratio) and the offset between the halo zone diameters and the agar hole diameters (SIoffset), respectively. The highest CMCase activity was 4.29 ± 0.071 U/ml. Morphological, physiological, biochemical, and 16S rRNA sequence analyses (100% homology with B. amyloliquefaciens sp. plantarum FZB42) were further carried out for the selected strain, leading to the identification of the strain as B. amyloliquefaciens sp. plantarum NDK5 strain. In addition, NDK5 was proved to have a capacity for synthesizing indole-3-acetic acid, a plant growth hormone, on an L-tryptophan-containing medium. Trial incubation of spent mushroom edible canna-substrate with the strain NDK5 showed increases in several quality criteria of the waste after 20 days of incubation, that meet the standard criteria for bio-organic fertilizer according to TCVN 7185:2002.

Prediction, enrichment, and isolation identify a responsive, competitive community of cellulolytic microorganisms from a municipal landfill

Landfills are engineered, heterogeneously contaminated sites containing large reservoirs of paper waste. Cellulose degradation is an important process within landfill microbial ecology, and these anoxic, saturated environments are prime locations for discovery of cellulases that may offer improvements on industrial cellulose degradation efforts. We sampled leachate from three locations within a municipal landfill, a leachate collection cistern, and groundwater from an adjacent aquifer to identify cellulolytic populations and their associated cellulases. Metagenomic sequencing identified wide-spread and taxonomically diverse cellulolytic potential, with a notable scarcity of predicted exocellulases. 16S rRNA amplicon sequencing detected nine landfill microorganisms enriched in a customized leachate medium amended with microcrystalline cellulose or common paper stocks. Paper-enrichment cultures showed three competition dynamics in response to the specific composition (lignin: hemi-cellulose: cellulose) of the different paper stocks. From leachate biomass, four novel cellulolytic bacteria were isolated, including two with the capacity for cellulolysis at industrially-relevant temperatures. None of the isolates demonstrated exocellulase activity, consistent with the metagenome-based predictions. However, there was very little overlap between metagenome-derived predicted cellulolytic organisms, organisms enriched on paper sources, or the isolates, suggesting the landfill cellulolytic community is at low abundance but able to rapidly respond to introduced substrates.