• Journal of Microbiology and Biotechnology
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• v.20 no.5
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• pp.889-892
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• 2010

We investigated a potential for glucose production from cellulose material using two kinds of hyperthermophilic enzymes, endocellulase (EG) and beta-glucosidase (BGL). Two BGLs, from hyperthermophile Pyrococcus furiosus and mesophile Aspergillus aculeatus, were compared with P. horikoshii endocellulase (EGPh) for complete hydrolysis of cellulose. The combination reactions by each BGL enzyme and EGPh could produce only glucose without the other oligosaccharides from phosphoric acid swollen Avicel (PSA). The combination of both the hyperthermophilic cellulases, BGLPf and EGPh, will be adaptable to a high efficiency system to produce glucose at high temperature.

• Plant Resources
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• v.8 no.3
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• pp.175-180
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• 2005

Thin sections of cellulose fibers were incubated with an endo- and an exoglucanase labeled with gold particles of differing sizes. The hydrolytic sites were then visualized under transmission electron microscopy (TEM). The potential interaction between the ${\beta}$-1, 4-glucan substrates and the endo- and the exoglucanases was investigated using cellulosic and lignocellulosic substrates. The simultaneous visualization was very successful in distinguishing preferred substrates for each cellulase in lignocellulosic substrates. When plant lignocellulose was preincubated with endocellulase, density of the gold labeling greatly increased suggesting that preliminary exposure of lignocellulosic material to endocellulase may have enhanced the accessibility of the substrate to endocellulase and exocellulase. This result provided a plausible explanation for the observed endo/exo cellulase co-hydrolysis.

• 한국당과학회:학술대회논문집
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• 2006.11a
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• pp.31-31
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• 2006

• Journal of Microbiology and Biotechnology
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• v.21 no.3
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• pp.284-292
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• 2011

An endocellulase-free multienzyme complex was produced by a thermophilic anaerobic bacterium, Thermoanaerobacterium thermosaccharolyticum strain NOI-1, when grown on xylan. The temperature and pH optima for growth were $60^{\circ}C$ and 6.0, respectively. The bacterial cells were found to adhere to insoluble xylan and Avicel. A scanning electron microscopy analysis showed the adhesion of xylan to the cells. An endocellulase-free multienzyme complex was isolated from the crude enzyme of strain NOI-1 by affinity purification on cellulose and Sephacryl S-300 gel filtration. The molecular mass of the multienzyme complex was estimated to be about 1,200 kDa. The multienzyme complex showed one protein on native PAGE, one xylanase on a native zymogram, 21 proteins on SDS-PAGE, and 5 xylanases on a SDS zymogram. The multienzyme complex consisted of xylanase, ${\beta}$-xylosidase, ${\alpha}$-L-arabinofuranosidase, ${\beta}$-glucosidase, and cellobiohydrolase. The multienzyme complex was effective in hydrolyzing xylan and corn hulls. This is the first report of an endocellulase-free multienzyme complex produced by a thermophilic anaerobic bacterium, T. thermosaccharolyticum strain NOI-1.

• Journal of Industrial Technology
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• v.29 no.A
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• pp.161-167
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• 2009

The bifunctional Xylanase-Cellulase hybrid protein was constructed by gene fusion. Two genes corresponding to endoxylanase gene (xylS) and endocellulase gene (celA) were amplified by PCR from Bacillus licleniformis NBL420. It was then linked through splicing by overlap extension (SOE) by PCR method. The two resulting fused hybrids, xyl/cel and cel/xyl, which differ by its orientation, were confirmed by its nucleotide sequencings. One of two fusion genes, xyl/cel was successfully expressed into pET22b(+) vector (pxyl/cel) with bifunctional xylanase-cellulase activity. On the contrary, the other cel/xyl fusion protein showed only cellulase activity with much decreased xylanase activity. Enzymatic properties of Xyl/Cel fusion protein were investigated regarding optimum pH, optimum temp, thermostability, and pH stability. It was revealed that Xyl/Cel fusion protein retained the bifunctional xylanase-cellulase activities eventhough two enzymes were connected with each other directly. These informations could be useful for construction of other hybrid proteins as well as increased range of substrate utilization.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.633-639
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• 1990

The genes for cellulases of Pseudomonas sp. LBC505 and CYC10, potent cellulase complex-producing strains, were cloned in Escherichia coli with pUC19. Recombinant plasmids pLCl and pLC2 were isolated from transformants producing cellulase by Congo red staining, and their genes cloned were 0.7 kb and 4.6 kb HindIII fragments, respectively. The inserts of pLCl and pLC2 were hybridized to chromosomal DNAs digested with HindIII from Pseudomona~ sp. LBC505 and CYC10, respectively. Immunodiffusion assays revealed that pLC1-and pLC2-encoded cellulase showed similarity with that of host strains. About 24% of cellulase activity was observed in the extracellular fraction of E. coli carrying pLC1, and its activity was higher about 1.4 times than that of LBC505. The enzymatic properties of pLC1 and pLC2 encoded cellulase were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases cloned were endocellulase.