• 한국토양동물학회지
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• 제3권2호
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• pp.51-57
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• 1998

A new rod-shaped endospore-forming bacterium isolated from the hindgut flora of the termite, Reticulitermes speratus kyushuenesis Morimoto is described. The isolate stained Gram positive, but the KOH test and the test for L-alanine aminopeptidase were negative. The length of a single cell varies from 2.5-9.0 $\mu $m, and the cell is about 0.5-0.7$\mu $m thick. The isolate had a high cellulolytic ability and was identified as Bacillus amyloliquefaciens.

• Journal of Microbiology and Biotechnology
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• 제27권3호
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• pp.573-583
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• 2017

Biofilm formation on the membrane surface results in the loss of permeability in membrane bioreactors (MBRs) for wastewater treatment. Studies have revealed that cellulose is not only produced by a number of bacterial species but also plays a key role during formation of their biofilm. Hence, in this study, cellulase was introduced to a MBR as a cellulose-induced biofilm control strategy. For practical application of cellulase to MBR, a cellulolytic (i.e., cellulase-producing) bacterium, Undibacterium sp. DM-1, was isolated from a lab-scale MBR for wastewater treatment. Prior to its application to MBR, it was confirmed that the cell-free supernatant of DM-1 was capable of inhibiting biofilm formation and of detaching the mature biofilm of activated sludge and cellulose-producing bacteria. This suggested that cellulase could be an effective anti-biofouling agent for MBRs used in wastewater treatment. Undibacterium sp. DM-1-entrapping beads (i.e., cellulolytic-beads) were applied to a continuous MBR to mitigate membrane biofouling 2.2-fold, compared with an MBR with vacant-beads as a control. Subsequent analysis of the cellulose content in the biofilm formed on the membrane surface revealed that this mitigation was associated with an approximately 30% reduction in cellulose by cellulolytic-beads in MBR.

• Journal of the Korean Wood Science and Technology
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• 제25권3호
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• pp.83-95
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• 1997

The degradation mode of lignocellulose by anaerobic ruminal cellulolytic bacterium Ruminococcus albus F-40 was investigated. Birchwood holocellulose and filter paper were incubated as the sole carbohydrate sources with using the Hungate techniques. After 2 or 4 days of incubation, samples were employed for chemical and electron microscopic evaluations. The degradation rate of cellulosic substrates and the adhesion rate of bacteria to the substrates increased proportionally with the decrease of relative crystallinity of cellulose, indicating the preferential breakdown of amorphous cellulose, by this bacterium. X-ray diffraction analyses and polarized light microscopy showed, however, that crystalline cellulose was also degraded by R. albus. FT-IR spectra indicated that not only cellulose but hemicellulose was also degraded by this bacterium. Electron microscopic investigations showed the protuberant structures on the surface of R. albus. These structures were much more significant when bacterial cells were grown in the media containing insoluble substrates, such as cellulose, indicating clearly that bacterial protuberant structures were induced by the substrates. Protuberant structures extended from the bacterial cells adhered tightly to the substrates and numerous vesicles covered the surface of cellulosic substrates affected. Cellulosome-like structures were distributed on the cellulose matrix. Electron microscopic works showed that diverse surface organells of R. albus were involved in the degradation of cellulosic materials. SEM examinations showed the breakdown of cellulose by R. albus was proceeded by severeal routes : short fiber formation, defibrillation and destrafication of cellulose microfibril.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1200-1210
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• 2004

Metabolic flux analysis was established by adapting previous stoichiometric model developed during growth with cellulose to cell grown with cellobiose for further direct comparison of the bacterial metabolism. In carbon limitation with cellobiose, a shift from acetate-ethanol fermentation to ethanol-lactate fermentation is observed and the pyruvate overflow is much higher than with cellulose. In nitrogen limitation with cellobiose, the cellodextrin and exopolysaccharide overflows are much higher than on cellulose. In carbon and nitrogen saturation with cellobiose, the cellodextrin, exopolysaccharide, and free amino acids overflows reach the highest levels observed but all remain limited on cellulose. By completely shunting the cellulosome, the use of cellobiose allows to reach much higher carbon consumption rates which, in return, highlights the metabolic limitation of C. cellulolyticum. Therefore, the physical nature of the carbon source has a profound impact on the metabolism of C. cellulolyticum and most probably of other cellulolytic bacteria. For cellulolytic bacteria, the use of soluble carbon substrate must carefully be taken into consideration for the interpretation of results. Direct comparison of metabolic flux analysis from cellobiose and cellulose revealed the importance of cellulosome, phosphoglucomutase and pyruvate-ferredoxin oxidoreductase in the distribution of carbon flow in the central metabolism. In the light of these findings, future directions for improvement of cellulose catabolism by this bacterium are discussed.

• Asian-Australasian Journal of Animal Sciences
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• 제14권10호
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• pp.1429-1433
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• 2001

This study provides electron microscopic evidence for the presence of cellulosome-like structures on the cell surface of Ruminococcus albus F-40. Electron microscopy showed that clusters of tightly packed spherical particles were located on the cell surface of R. albus. The protuberant structures present mainly on the bacterial surface and also bound to the cellulose substrate appeared to be the site of cellulosome-like structures. From the evidence presented, we suggest that the structures described here might be a characteristic feature of some ruminal cellulolytic bacteria.

• Journal of Microbiology and Biotechnology
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• 제17권11호
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• pp.1789-1796
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• 2007

A ${\beta}$-glucosidase with the molecular mass of 160,000 Da was purified to homogeneity from cell extract of a cellulolytic bacterium, Cellulomonas uda CS1-1. The kinetic parameters ($K_m$ and $V_{max}$) of the enzyme were determined with pNP-cellooligosccharides (DP 1-5) and cellobiose. The molecular orbital theoretical studies on the cellulolytic reactivity between the pNP-cellooligosaccharides as substrate (S) molecules and the purified ${\beta}$-glucosidase (E) were conducted by applying the frontier molecular orbital (FMO) interaction theory. The results of the FMO interaction between E and S molecules verified that the first stage of the reaction was induced by exocyclic cleavage, which occurred in an electrophilic reaction based on a strong charge-controlled reaction between the highest occupied molecular orbital (HOMO) energy of the S molecule and the lowest occupied molecular orbital (LUMO) energy of the hydronium ion ($H_3O^+$), more than endocyclic cleavage, whereas a nucleophilic substitution reaction was induced by an orbital-controlled reaction between the LUMO energy of the oxonium ion ($SH^+$) protonated to the S molecule and the HOMO energy of the $H_2O_2$ molecule. A hypothetic reaction route was proposed with the experimental results in which the enzymatic acid-catalyst hydrolysis reaction of E and S molecules would be progressed via $SN_1$ and $SN_2$ reactions. In addition, the quantitative structure-activity relationships (QSARs) between these kinetic parameters showed that $K_m$ has a significant correlation with hydrophobicity (logP), and specific activity has with dipole moment, respectively.

• Journal of Microbiology and Biotechnology
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• 제20권5호
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• pp.893-903
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• 2010

A cellulolytic and xylanolytic enzyme complex-producing alkalothermoanaerobacterium strain, Tepidimicrobium xylanilyticum BT14, is described. The cell was Grampositive, rod-shaped, and endospore-forming. Based on 16S rRNA gene analysis and various lines of biochemical and physiological properties, the strain BT14 is a new member of the genus Tepidimicrobium. The strain BT14 cells had the ability to bind to Avicel, xylan, and corn hull. The pH and temperature optima for growth were 9.0 and $60^{\circ}C$, respectively. The strain BT14 was able to use a variety of carbon sources. When the bacterium was grown on corn hulls under an anaerobic condition, a cellulolytic and xylanolytic enzyme complex was produced. Crude enzyme containing cellulase and xylanase of the strain BT14 was active in broad ranges of pH and temperature. The optimum conditions for cellulase and xylanase activities were pH 8.0 and 9.0 at $60^{\circ}C$, respectively. The crude enzyme had the ability to bind to Avicel and xylan. The analysis of native-PAGE and native-zymograms indicated the cellulosebinding protein showing both cellulase and xylanase activities, whereas SDS-PAGE zymograms showed 4 bands of cellulases and 5 bands of xylanases. Evidence of a cohesinlike amino acid sequence seemed to indicate that the protein complex shared a direct relationship with the cellulosome of Clostridium thermocellum. The crude enzyme from the strain BT14 showed effective degradation of plant biomass. When grown on corn hulls at pH 9.0 and $60^{\circ}C$ under anaerobic conditions, the strain BT14 produced ethanol and acetate as the main fermentation products.

• 한국미생물·생명공학회지
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• 제46권1호
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• pp.59-67
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• 2018

A xylanolytic and cellulolytic anaerobic bacterium strain CtC72 was isolated from cattle rumen liquor. Phylogenetic analysis of 16S rRNA gene sequences revealed that strain CtC72 shared only 97.78% homology with its nearest phylogenetic affiliate Actinomyces ruminicola, showing its novelty. The strain could grow on medium containing xylan, carboxymethyl cellulose and avicel producing $CO_2$, acetate, and ethanol as major fermentation products. The whole genome analysis of the strain CtC72 exhibited a broad range of carbohydrate-active enzymes required for the breakdown and utilization of lignocellulosic biomass. Genes related to the production of ethanol and stress tolerance were also detected. Further there were several unique genes in CtC72 for chitin degradation, pectin utilization, sugar utilization, and stress response in comparison with Actinomyces ruminicola. The results show that the strain CtC72, a putative novel bacterium can be used for lignocellulosic biomass based biotechnological applications.

• Asian-Australasian Journal of Animal Sciences
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• 제20권5호
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• pp.802-810
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• 2007

Fibrobacter succinogenes, a Gram-negative, anaerobic ruminal bacterium is a major fibre digesting species in the rumen. It intensively degrades plant cell walls by an erosion type of mechanism, burrowing its way through the complex matrix of cellulose and hemicellulose with the release of digestible and undigested cell wall fragments. The enzymes involved in this process include a combination of glucanases, xylanases, arabinofuranosidase(s) and esterases. The genome of the bacterium has been sequenced and this has revealed in excess of 100 putative glycosyl hydrolase, pectate lyase and carbohydrate esterase genes, which is greater than the numbers reported present in other major cellulolytic organisms for which genomes have been sequenced. Modelling of the amino acid sequences of two glycanases, CedA and EGB, by reference to crystallized homologs has enabled prediction of the major features of their tertiary structures. Two dimensional gel electrophoresis in conjunction with mass spectroscopy has permitted the documentation of proteins over expressed in F. succinogenes grown on cellulose, and analysis of the cell surfaces of mutant strains unable to bind to cellulose has enabled the identification of candidate proteins with roles in adhesion to the plant cell wall substrate, the precursor to cellulose biodegradation.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1280-1290
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• 2014

In order to obtain the cellulolytic bacterial consortia, sediments from Great Basin hot springs (Nevada, USA) were sampled and enriched with cellulosic biomass as the sole carbon source. The bacterial composition of the resulting anaerobic ethanol-producing celluloytic bacterial consortium, named SV79, was analyzed. With methods of the full-length 16S rRNA library-based analysis and denaturing gradient gel electrophoresis, 21 bacteria belonging to eight genera were detected from this consortium. Clones with closest relation to the genera Acetivibrio, Clostridium, Cellulosilyticum, Ruminococcus, and Sporomusa were predominant. The cellulase activities and ethanol productions of consortium SV79 using different agricultural residues (sugarcane bagasse and spent mushroom substrate) and energy crops (Spartina anglica, Miscanthus floridulus, and Pennisetum sinese Roxb) were studied. During cultivation, consortium SV79 produced the maximum filter paper activity (FPase, 9.41 U/ml), carboxymethylcellulase activity (CMCase, 6.35 U/ml), and xylanase activity (4.28 U/ml) with sugarcane bagasse, spent mushroom substrate, and S. anglica, respectively. The ethanol production using M. floridulus as substrate was up to 2.63 mM ethanol/g using gas chromatography analysis. It has high potential to be a new candidate for producing ethanol with cellulosic biomass under anoxic conditions in natural environments.