• Journal of Industrial Technology
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• v.29 no.B
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• pp.121-125
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• 2009

New cellulase genes, named as CelV2 and CelN1, respectively, were isolated from Erwinia carotovora ATCC15713 and expressed in E. coli. The CelV2 and CelN1 gene were PCR amplified with degenerated primers and PCR products were sequenced and expressed in E. coli. Two new cellulase genes showed 97% homologies with previously reported Erwinia cellulase genes. The recombinant cellulase were purified with Ni-NTA column chromatography and its enzymatic properties were characterized. The optimum temperature of two enzymes were about $50^{\circ}C$ degree and optimum pH were around pH7.0. The newly isolated celluase genes could be used for enhancing substrate range of alcohol-producing bacteria such as Zymomonas mobilis.

• Microbiology and Biotechnology Letters
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• v.15 no.5
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• pp.346-351
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• 1987

A cellulase gene of Clostridium themocellum was transferred to Escherichia coli by molecular cloning with pBR322. The gene was carried in a Hind III digested DNA sequence of about 1.8 kb. This Rind III fragment expressed activities on carboxymethyl cellulose (CMC) and on filter gaper in E. coli. The expression of clostridial cellulase gene in E. coli was studied and compared with the pro-ducts of cellulase genes in C. themocellum.

• Journal of Life Science
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• v.22 no.7
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• pp.912-919
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• 2012

A Bacillus sp. strain producing celluase and xylanase was isolated from environmental soil with LB agar plate containing carboxymethylcellulose (CM-cellulose) and beechwood xylan stained with trypan blue as substrates, respectively. Based on the 16S rRNA gene sequence and API 50 CHL test, the strain was identified as B. subtilis and named B. subtilis NC1. The cellulase and xylanase from B. subtilis NC1 exhibited the highest activities for CM-cellulose and beechwood xylan as substrate, respectively, and both enzymes showed the maximum activity at pH 5.0 and $50^{\circ}C$. We cloned and sequenced the genes for cellulase and xylanase from genomic DNA of the B. subtilis NC1 by the shot-gun cloning method. The cloned cellulase and xylanase genes consisted of a 1,500 bp open reading frame (ORF) encoding a 499 amino acid protein with a calculated molecular mass of 55,251 Da and a 1,269 bp ORF encoding a 422 amino acid protein with a calculated molecular mass of 47,423 Da, respectively. The deduced amino acid sequences from the genes of cellulase and xylanase showed high identity with glycosyl hydrolases family (GH) 5 and 30, respectively.

• 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.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.524.1-524
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• 1986

CMCase, a member of cellulose decomposing enzymes, hydrolyze cellulose up to cellobiose. Cellobiase splits cellobiose to glucose units. Therefore, a linkage of the twogenes coding for CMCase and cellobiase on the same plasmid is needed to produce a cellulase complex which can produce glucose from cellulose. A genetic operon in which the two structural genes are under the control of a single promoter would be ideal for this purpose. The present report is on the linking of the two cellulase genes in one plasmid as a preliminary step of the operon construction.

• 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.

• Journal of Animal Science and Technology
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• v.62 no.5
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• pp.761-763
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• 2020

Here we report the complete genome sequence of Bacillus amyloliquefaciens ATC6, which produces acidic cellulase, isolated from pig feces. The genome is 4,062,817 bp in length and has a guanine-cytosine (GC) content of 46.27%. Among the predicted 3,913 protein-coding genes, two glucanase genes, which are involved in lichenan and cellulose degradation, were found. This genome analysis helps clarify the mechanism involved in cellulose biodegradation and support its application for efficient use of livestock feeds.

• Journal of Microbiology and Biotechnology
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• v.22 no.8
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• pp.1133-1140
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• 2012

Ribonucleic acid interference (RNAi) inhibits the expression of target genes in a sequence-specific manner, and shows potential for gene knockdown in filamentous fungi, in which the locus-specific gene knockout occurs in low frequency. In this study, the function of the repressor of cellulase expression I (ACEI) was verified in Trichoderma koningii (T. koningii) YC01 through RNAi, and ace1-silenced strains with improved cellulase productivity were obtained. An expression cassette that transcribed the interfering double-stranded RNA (dsRNA) of ace1 was constructed and transformed into T. koningii, and the transformants, in which the expression of ace1 was successfully silenced, were selected. As a result of the ace1 gene silencing, the expression levels of the main cellulase and xylanase genes were elevated, and the enhanced production of total proteins, cellulase, and xylanase was observed in the cultivation. In addition, the down-regulation of ace1 resulted in an increasing expression of xyr1, but no clear variation in the expression of cre1, which suggested that ACEI acted as a repressor of the xyr1 transcription, but was not involved in the regulation of the cre1 expression. The results of this work indicate that ace1 is a valid target gene for enhancing enzyme production in T. koningii, and RNAi is an appropriate tool for improving the properties of industrial fungi.

• Microbiology and Biotechnology Letters
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• v.39 no.1
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• pp.20-28
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• 2011

Phylogenetic analysis of two cellulase genes, xynB1 and bglA2, and the groEL1 gene from 34 Sorangium cellulosum strains isolated in Korea suggested that there are at least five subgroups in S. cellulosum, which is the most proficient producer of secondary metabolites among myxobacteria. This analysis also revealed diversity among the isolated S. cellulosum. It appeared that at least 30 out of 34 strains are different each other.

• Korean Journal Plant Pathology
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• v.13 no.3
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• pp.172-178
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• 1997

Xanthmonas campestris pv. campestris, causal agent of Black rot of crucifers, were isolated and identified from crucifer host. In order to determine the characters of X. c. pv. campestris associated with pathogenicity, Tn5 mutagenesis was carried out by conjugating with E. coli pJB4J1. Transconjugants were plate- assayed for missing cellulase, protease and amylase activity. A cellulase negative mutant was selected and tested for pathogenicity. Light microscopy and Scanning electron microscopy revealed that substomatal tissues were colonized by mutant, but was far less extensive than those by wild type. Stomatal surface and substomatal tissue appeared to have degraded by only wild type in 24 hrs and progression of pathogenesis was distinct in 48 hrs. In 6 days, wild type proliferated well in the tissue facilitated by cellulase activity. As a result, cellulase was determined as the important factor in pathogenesis.