• Journal of Microbiology and Biotechnology
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• v.20 no.5
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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.

• KSBB Journal
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• v.11 no.6
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• pp.712-717
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• 1996

Cellulase was modified with synthetic copolymers of polyoxyethylene derivative and maleic acid anhydride. The saccharification characteristics and enzymatic reaction kinetic mechanism of modified and native cellulases were observed. In modification reaction of cellulase, degree of modification(DM) increased, as mass ratio of copolymers to enzyme increased. Maximum DM was 55% at mass ratio of 4 and remained activity was 75%. In saccharification experiment modified enzyme had maintained higher stability than native enzyme over all the reaction and the final conversion yield of modified enzyme was greater than that of native enzyme. Numerical simulation based on the reaction mechanism considering enzymatic deactivation was performed. Modified enzyme had kept higher free enzyme concentration over all the reaction than that of native enzyme. Comparing calculation values with experimental data, calculation values were in accordance with experimental data.

• Applied Biological Chemistry
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• v.38 no.6
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• pp.490-495
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• 1995

Among the cellulases by Cellulomonas sp. KL-6. CMCase and filter paperase, which were produced as the out enzymes of cell, had been much produced, but very small amounts of ${\beta}-glucosidase $, the enzyme of which is cell bound form, was produced by this organism. The optimal culture times for CMCase and filter paperase productions were 5 days, while that of ${\beta}-glucosidase$ was 4 days. When this strain was cultured under the optimal medium for enzyme production, CMCase, FPase and ${\beta}-glucosidase$ were $82\;units/m{\ell},\;80\;units/m{\ell}\;and\;1.2\;units/m{\ell}$, respectively. Thus these results were showed to increase enzyme productivities as about $60{\sim}70%$ than those produced in basal medium. $CaCO_3$ injected to the medium as the ratio of 0.1% was not only enhanced cellulase activities but also effective as acid neutralizing agent. The production effects of lignase and lactase by this bacterium in filter paper medium was not appeared.

• Journal of Physiology & Pathology in Korean Medicine
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• v.26 no.1
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• pp.35-39
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• 2012

Sweet potato soju(SPS) has been made by vacuum distillation because sweet potato contains much fibrous materials which give high density to sweet potato mash. Generally, the SPS made by vacuum distillation has soft flavors and tastes. If the viscosity of sweet potato mash could be decreased by degradation enzyme, the process and production of SPS making by the method of vacuum distillation may be simplified and easier to distil the fermented sweet potato. Because the fibrous materials of sweet potato contains pectin with methoxyl group, methanol can be produced by fibrous degradation enzyme. For appling the fiber degradation enzymes to sweet potato mash for making SPS, the enzyme should be needed to degrade fibrous material without producing methanol. Special two fibrolytic enzymes are selected from 26 kind of commercial enzymes for the simplified and easier production of sweet potato soju by vacuum distillation, The selected enzyme A and X can degrade the fibrous material pectin of sweet potato without producing methanol. Although the different companies have produced the enzymes, same cellulase has been prepared from Trichoderma. reesei. The viscosity of sweet potato mash treated by the enzymes is decreased by 3 times with comparison to the viscosity of sweet potato mash of control group. The methanol concentration in the vacuum distilled SPS treated with the enzymes is 0.16%. The concentration is similar to that of commercially distilled SPS(0.15%). The result may suggest that the selected cellulases, A and X, can be used to make SPS by vacuum distillation.

• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.539-546
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• 2020

A cellulolytic bacterial strain, S2-3, was isolated from sea water collected in Jeju island, Republic of Korea. The strain was aerobic and gram negative, and formed yellow colored colonies on marine agar medium. S2-3 cells were long rod-shaped, 0.5 × 0.25 ㎛ (width x length) in size, and did not have flagella. The optimal growth conditions for S2-3 were 30-35℃ and pH 6.5-7.0. Analysis of the 16S rRNA gene sequence of S2-3 revealed that it had the highest identity with those of Seonamhaeicola algicola Gy8 (97.08%), Hyunsoonleella udonensis JG48 (95.01%), and Aestuariibaculum scopimerae I-15 (94.86%). In phylogenetic analysis, S2-3 formed the same clade as S. algicola Gy8, implying that S2-3 belongs to the genus Seonamhaeicola. The major fatty acids (>10%) comprised C15:1 iso G (22.29%), C15:0 iso (17.71%), C17:0 iso 3OH (16.06%), and C15:0 iso 3OH (10.7%), resulting in quite different ratio of the component from those of S. algicola Gy8. Moreover, its biochemical characteristics, including acid production and enzyme activities, were different from those of S. algicola Gy8. Therefore, putting all these results together, we concluded S2-3 is distinct species from S. algicola Gy8, and thus named it Seonamhaeicola sp. S2-3. In liquid culture, S2-3 produced extracellular cellulases that can hydrolyze cellulose or cellooligosaccharides into cellobiose, which is a good enzyme resource that deserves further research.

• Microbiology and Biotechnology Letters
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• v.50 no.2
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• pp.245-254
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• 2022

Cellulases are a group of biocatalyst enzymes that are capable of degrading cellulosic biomass present in the natural environment and produced by a large number of microorganisms, including bacteria and fungi, etc. In the current study, we isolated, screened and characterized cellulase-producing bacteria from soil. Three cellulose-degrading species were isolated based on clear zone using Congo red stain on carboxymethyl cellulose (CMC) agar plates. These bacterial isolates, named as HB2, HS5 and HS9, were subsequently characterized by morphological and biochemical tests as well as 16S rRNA gene sequencing. Based on 16S rRNA analysis, the bacterial isolates were identified as Bacillus cerus, Bacillus subtilis and Bacillus stratosphericus. Moreover, for maximum cellulase production, different growth parameters were optimized. Maximum optical density for growth was also noted at pH 7.0 for 48 h for all three isolates. Optical density was high for all three isolates using meat extract as a nitrogen source for 48 h. The pH profile of all three strains was quite similar but the maximum enzyme activity was observed at pH 7.0. Maximum cellulase production by all three bacterial isolates was noted when using lactose as a carbon rather than nitrogen and peptone. Further studies are needed for identification of new isolates in this region having maximum cellulolytic activity. Our findings indicate that this enzyme has various potential industrial applications.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.68-72
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• 2010

The ${\beta}$-1,4-glucanase gene of Bacillus licheniformis B1 was expressed in Esherichia coli BL21, and a protein with a mass of 50 kDa that was soluble was overproduced. A protein with a mass of 37 kDa was secreted from B. licheniformis. It seems that the ${\beta}$-1,4-glucanase produced in E. coli contained the leader peptide and unprocessed carboxy-terminal region, but its processing occurred in the carboxyterminal in Bacillus. The optimal temperature of ${\beta}$-1,4-glucanase was $40^{\circ}C$. The enzyme still had 76% maximal activity at $60^{\circ}C$. The optimal pH of the enzyme was 7. The enzyme retained considerable activities over the weak-acidic, neutral, and weak-basic pH range. Acidic fungal cellulases are used in food, detergent, pulp, paper, textile industries. However, studies about neutral and alkaline cellulase are not enough. The cellulase developed in this study may be useful for industrial applications in the fields of biofuel development.

• Journal of the Korea Organic Resources Recycling Association
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• v.15 no.3
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• pp.80-89
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• 2007

Two endogenous endo-${\beta}$-1,4-D-glucanase (EGase, EC 3.2.1.4) cDNAs were cloned from the midgut of the earthworm Eisenia anderi, and named EaEG2 and EaEG3, respectively. A sequence of 1,368 bp was determined and the coding region is composed of 456 amino acid residues including the initiation methionine. The N-terminal region of 20 residues in the deduced sequence was regarded as the signal peptide. These EGases belong to glycosyl hydrolase family 9 (GHF9) and showed high levels of identity(51-55%) with selected termite, cockroache, crayfish and mollusc EGases. The EGases of earthworm consist of three consensus catalytic domains found in most microbial cellulases. A phylogenetic tree was constructed using the deduced amino acid sequence data matched through the BLASTX program and showed that GHF9 families could be divided into five groups of arthropoda, bacteria, plant, annelida and mollusc.

• Journal of Animal Science and Technology
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• v.45 no.4
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• pp.659-666
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• 2003

Endoglucanase A from Clostridium thermocellum which is resistant to pancreatic proteinase was selected out of numbers cellulases then were expressed in lactobacilli. Recombinant lactobacilli expression vector, pSD1, harboring the endoglucanase gene from C. thermocellum under the control of its own promoter, was constructed. Both L. bulgaricus and L. plantarum were electrotransformed with pSD1. The endoglucanase activities of 0.120 and 0.144 U/ml were found in culture media of L. bulgaricus and L. plantarum containing pSD1, respectively. In vitro survival characteristics of the transformed lactobacilli were tested. Both L. bulgaricus and L. plantarum showed a similar resistance to low pH 3. Moreover, L. plantarum was bile-salt resistant in the presence of 0.3 and 1% oxgall. L. bulgaricus and L. plantarum showed a rather homogenous resistant pattern against the tested antibiotics. Both of the strains were resistant to amikacin, gentamicin, streptomycin, kanamycin, and colistin.

• Korean Journal of Food Science and Technology
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• v.23 no.1
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• pp.31-36
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• 1991

Cellulase genes from thermophilic alkalophilic Bacillus sp. F204 a potent cellulase complex-producing bacterium, were cloned in Escherichia coli with pUC 19. Plasmids pBC191 and pBC192, isolated from transformants forming yellow zone around colony on the LB agar plate containing 0.5% carboxymethyl cellulose and ampicillin, contained 4.6 Kb and 5.8 Kb HindIII fragments, respectively. The 4.6 Kb insert of pBC191 had single sites for BamHI EcoRI, KpnI and pvuII. DNA hybridization and immunodiffusion studies showed that pBC191-encoded cellulase gene was homologous with that of host strain. pKC231, constructed by inserting 4.6 Kb insert of pBC191 at the HindIII site of pKK223-3, E. coli expression vector, and pGC711, constructed by inserting 4.6 Kb insert of pBC191 at the HindIII site of pGR71, E. coli and B. subtilis shuttle vector, had 3.2 times and 2.8 times as much cellulase activity as pBC191, respectively. Substrate specificity analysis showed that cellulases cloned were CMCase.