• 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.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.41-47
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• 2004

This study was performed to investigate the antimicrobial effects of hetero-chitosans and their oligosaccharides on the halophilic bacterium, Vibrio parahaemolyticus. Nine classes of hetero-chitosan oligosaccharides were prepared based on their molecular weights, using an ultrafiltration membrane reactor system with chitosanase and celluase, from partially different deacetylated chitosans, 90%, 75%, and 50% deacetylated chitosan, respectively. Thirty-two strains of V. parahaemolyticus were isolated from various marine organisms such as shellfish, shrimps, octopus, and seabirds. Seventy-five percent deacetylated chitosan showed the highest antimicrobial acitivity. The minimal inhibitory concentration (MIC) was 0.5 mg/ml on 14 strains of V. parahaemolyticus, and MIC of the rest strains (18 strains) was 1.0 mg/ml. In addition, MIC of most hetero-chitosan oligosaccharides was 8.0 mg/ml. The results revealed that the antimicrobial effects of hetero-chitosans and their oligosaccharides against V. parahaemolyticus depend on the degree of deacetylation, their molecular weights, and strains tested.

• Korean Journal of Pharmacognosy
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• v.20 no.1
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• pp.43-47
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• 1989

An attempt was made to increase the yield of extraction of ginkgoflavonglycosides from leaves of Ginkgo biloba by treatments of with Cellulase C and macerating enzymes. The yield of dried extract and its contents of ginkgoflavonols, when treated only with cellulase C, were analyzed to be 1. 99% and 0. 38%, respectively. The contents of ginkgoflavonglycosides in the dried extracts were calculated to be 25. 28%. By the treatment with a mixture of three enzymes, cellulase C: cellulase NC and macerosin (1 : 1 : 2), the yield of the dried extract, ginkgoflavonols as well as their glycosides were determined to be 2. 48%, 0. 48% and 24. 16%, respectively.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 2003.10a
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• pp.154-154
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• 2003

국내산 홍화씨를 부위별(Whole, Coat and Endosperm)로 분리한 후 추출용매, 볶음조건, 효소가수분해 등의 처리조건을 달리하여 추출된 추출물의 이화학적특성과 기능성성분 함량을 분석, 비교하였다. 홍화씨 부위별추출물의 고형분함량은 60%에탄올배유부분추출물이 11.29%로, 19$0^{\circ}C$, 30분 볶음처리배유추출물이 14.53%로, amyloglucosidase 처리배유추출물이 24.21%로 높은 고형분이 추출되었다. 총페놀함량은 추출용매의 에탄올농도가 증가함에 따라 증가하는 경향이었고, 80% 에탄올점질추출물이 965mg%로 가장 높았으며, 21$0^{\circ}C$, 30분 볶음처리껍질추출물이 756 mg%로, celluase처리껍질추출물이 1170mg%로 높은 함량이었다. 총플라보노이드함량은 80%에탄올전체추출물이 317 mg%로, 21$0^{\circ}C$, 30분 볶음 처리배유추출물이 488 mg%로, $\beta$-amylase 처리전체추출물이 554 mg%로 높은 함량이었다. 유리당 중 sucrose함량은 21$0^{\circ}C$, 10분 볶음처리배유추출물이 123.4 mg%로, 60%에탄올배유추출물이 57.0 mg%로, Celluase 처리배유추출물이 67.1 mg%로 가장 높은 함량이었고, 또한 glucose, fructose, xylose 및 arabinose 등도 함유하고 있었다. 유기산 중 citric acid 함량은 20%n 에탄올배유추출물이 243.3 67.1 mg%로, 21$0^{\circ}C$, 30분 볶음처리전체추출물이 76.3 mg%로, amyloglucosidase 처리배유추출물이 699.3 mg%로 가장 높은 함량이었고, 또한, oxalic, malic, succinic, acetic 및 fumaric acid 등도 확인되었다. Serotonin 화합물 중 serotonin- I 함량은 100% 에탄올껍질추출물이 431 mg%로, 21$0^{\circ}C$, 10분 볶음처리껍질추출물이 192 mg%로, amyloglucosidase 처리껍질추출물이 256 mg%로 가장 많았다. 또한, Serotonin-II함량은 100%에탄올껍질추출물이 763 mg%로, 17$0^{\circ}C$, 10분 볶음처리전체추출물이 312 mg%로, amyloglucosidase 처리껍질추출물이 456 mg%로 가장 많았다. Acacetin 함량은 80%에탄올배유추출물이 34.9 mg%로, 21$0^{\circ}C$, 30분 볶음처리배유추출물이 221.0 mg%로, amyloglucosidase 처리배유추출물이 27.8 mg%로 가장 많았다.

• 한국신재생에너지학회:학술대회논문집
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• 2010.06a
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• pp.253-253
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• 2010

Rice straw was pretreated with aqueous ammonia in order to enhance enzyme digestibility. Soaking in ammonia aqueous (SAA) was conducted with 15% ammonia, at $60^{\circ}C$. for 24 h. Optimization of both saccharification conditions and enzyme loading of SAA rice straw was carried out. Especially enzyme loading test was performed using statistical method. Moreover proton beam irradiation (PBI) was also performed to overcome the problem which inhibit the enzyme digestibility at 1-25 kGy doses with 45 MeV of beam energy. Optimal condition for enzymatic saccharification was follows; pH 4.8, $50^{\circ}C$, 60 FPU of enzyme activity, 1:4 ratio of celluase and ${\beta}$-glucosidase. Also, optimal doses of PBI on rice straw and SAA-treated rice straw for efficient sugar recovery were found to be 3 kGy, respectively. When saccharification was performed with optimal condition, glucose conversion yield was 89% of theocratical maximum in 48 h, and 3 kGy of PBI was applied to SAA-treated rice straw, approximately 90% of the theoretical glucose yield was obtained in 12 h. The results of X-ray diffractometry (XRD) support the effect of both SAA and PBI on sugar recovery, and scanning electron microscopy (SEM) images unveiled the physical change of the rice straw surface since rugged rice straw surface was observed.

• Korean Journal of Microbiology
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• v.45 no.3
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• pp.257-262
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• 2009

Carboxymethyl celluase (cellulase) was purified from cell-free extract of the recombinant Escherichia coli carrying a Bacillus licheniformis WL-12 cellulase gene by DEAE-Sepharose and phenyl-Sepharose column chromatography with specific activity of 163 U/mg protein. The molecular mass of the purified enzyme was estimated to be approximately 49.5 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme had a pH optimum at 5.5 and a temperature optimum at $55^{\circ}C$. The activity of the enzyme was completely inhibited by SDS (5 mM), and slightly enhanced by $Cu^{2+}$ (5 mM). The cellulase was active on CMC, konjac, barely glucan and lichenan, while it did not exhibit activity towards xylan, locust bean gum, and p-nitrophenyl-$\beta$-glucopyranoside. The predominant products resulting from the cellulase hydrolysis were cellobiose and cellotriose for cellooligosaccharides including cellotriose, cellotetraose and cellopentaose. The enzyme could hydrolyze cellooligosaccharides larger than cellobiose.

• 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.

• The Journal of Natural Sciences
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• v.4
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• pp.103-142
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• 1991

These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

• 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 the Korean Applied Science and Technology
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• v.35 no.3
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• pp.702-708
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• 2018

Hydrogels are natural polymer networks that can contain huge quantities of water and many cosmetical ingredients. Their hydrophilic functional groups creates a matrix, which allows high efficacy in delivering active ingredients into the skin. In industry, hydrating properties and strength of the hydrogels are of great interest in manufacturing hydrogel mask packs. We have used the cellulose in various forms such as powder, cotton fiber and cellulase treated cotton fiber to investigate the property changes of cellulose/hydrogel sheets. When 0.1% and 0.3% of cellulose powder were added to hydrogels, tensile strength of hydrogel sheets were decreased by 10% and 14% respectively. Vise versa, when 0.5 ~ 2 cm of cotton fibers were added, tensile strength of hydrogel sheets were significantly increased by about 20%. The hydrogels which contain cotton fibers also gave an excellent moisturizing effect. Especially cellolose/hydrogels containing cellulase-treated cotton fibers showed the best effect on retaining moisture content increasing upto 380% in comparison with the one containing untreated cotton as well as excellent dispersibility.