• Title/Summary/Keyword: 섬유소분해 활성

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Effects of Anthriscus sylvestris Hoffmann Extract on the Biosynthesis of Collagen in Human Dermal Fibroblasts (사람 섬유아세포에서 전호(Anthriscus sylvestris Hoffmann)추출물이 콜라겐 생합성에 미치는 영향)

  • Lee, Woo-Jung;Kim, Yong-Kee;Kim, Su-Nam
    • Korean Journal of Plant Resources
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    • v.25 no.2
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    • pp.240-245
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    • 2012
  • In this study, we investigated the boosting effects on collagen biosynthesis of $Anthriscus$ $sylvestris$ ethanol extract (ASE) in human dermal fibroblasts. To obtain more effective fraction and subfraction for collagen biosynthesis, standard solvent partition and open column chromatography were performed. The EtOH extract, solvent fractions, and 8 EtOAc subfractions were tested for their collagen synthesis capacity by [$^3H$]Proline-incorporation and ELISA assay. ASE increased 25% of total collagen synthesis and 27% of procollagen biosynthesis. The total collagen biosynthesis was increased by EtOAc fraction and E6 subfraction to 28% and 50% respectively. Type I procollagens were also upregulated by EtOAc fraction and E6 subfraction to 30% and 47%, each. Taken together, our data suggest that potential anti-aging effect of ASE on skin is via increasing collagen biosynthesis and effective subtraction is E6 subfraction of EtOAc fraction.

Characteristics of Fibrinolytic Enzymes of Bacillus licheniformis CY-24 Isolated from Button Mushroom Compost (양송이 배지로부터 분리한 Bacillus licheniformis CY-24의 섬유소분해 효소의 특성)

  • Min, Gyeong-Jin;Park, Hea-sung;Lee, Een-ji;Lee, Chan-Jung
    • The Korean Journal of Mycology
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    • v.49 no.2
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    • pp.199-209
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    • 2021
  • The present study was performed to improve the technique used for fermenting the mushroom growth medium. Taxonomic analysis of 16S rDNA sequence from the predominant Bacillus strain CY-24 isolated during the fermentation phase of the rice straw medium identified it as Bacillus licheniformis. In addition, the growth environment of B. licheniformis was also examined in this study, which revealed the optimal growth temperature and pH to be 30 ℃ and 6.0, respectively. This study also revealed that carboxymethyl cellulase (CMCase) and polygalacturonase (PGase) enzymes isolated from B. licheniformis achieved their maximal activities at 50 ℃ and 60 ℃ respectively. Furthermore, the study confirmed that the two enzymes, i.e., CMCase and PGase in B. licheniformis are stable at temperatures above 60 ℃. The present study thus demonstrates that B. licheniformis CY-24 possesses excellent enzymatic properties. It also reveals that the action of enzymes during the production of growth mediums used for the cultivation of mushrooms is closely associated with the promotion of fermentation and softening of the rice straw. Overall, this study provides elementary information regarding the role of B. licheniformis enzymes during growth medium fermentation for Agaricus bisporus cultivation.

Bacterial Strains and Their Cellulase Activity from the Excavated Clothes at Daedeok-gu, Daejeon (대전 대덕구 출토 복식 유물에 부착된 세균의 종류 및 섬유소분해효소 활성)

  • Lee, Sang-Joon;Cha, Mi-Sun;Cho, Hyun-Hok;Back, Young-Mee;Kwon, Young-Suk
    • Fashion & Textile Research Journal
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    • v.7 no.1
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    • pp.70-74
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    • 2005
  • This study was aimed to isolation and identification of attached bacteria on the clothes excavated from Deajeon area dating on the 16th century. From the observation with colony shape, 17 bacterial strains were isolated, and then 7 bacterial strains were identified with morphological and biochemical characteristics. Streptococcus sp., Alcaligenes faecalis, Gemella sp., Acinetobacter sp., Pseudomonas vesicularis, Aeromonas sain. salmonicida, Moraxella spp. In observation of the bacterial strains by the sort of textile, more bacterial strains were found in silk, cotton, and cotton batt than in ramie and hemp. It is suggest that hemp has antibacterial characteristics due to the presence of lignin. In the comparison washed samples with unwashed ones, there were more kinds of bacterial strains in washed samples. In the cellulase activity tests, all isolated bacteria had low level cellulase activity.

Cloning and Characterization of a Cellulase Gene from a Plant Growth Promoting Rhizobacterium, Bacillus subtilis AH18 against Phytophthora Blight Disease in Red-Pepper (고추역병을 방제하는 PGPR균주 Bacillus subtilis AH18의 항진균성 Cellulase 유전자의 Cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Jung, Hee-Kyoung;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.311-317
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    • 2006
  • Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

Characterization of Carboxymethyl Cellulase Produced by Cellulomonas sp. CS1-1 on Microcrystalline Cellulose (Cellulomonas sp. CS1-1이 미소결정성 섬유소로부터 생산한 Carboxymethyl Cellulase의 효소적 성질)

  • Park, Jong-Soo;Yoon, Min-Ho;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.24 no.2
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    • pp.275-282
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    • 1997
  • The prcx.iuction of extracellular 1,4-${\beta}$-glucanase by Cellulomonas sp. CS1-1 on microcrystalline cellulose, sigmacell was maximal after 5-day cultivation as 280 units/mL, which was three times higher than the level produced on carboxymethyl cellulose. A carboxymethyl cellulase containing the carbohydrate of 8.2% was purified from the culture filtrate by successive procedures of column chromatographies. Purification factor was calculated as 22-folds with the specific carboxymethyl cellulase activity of 31.9 units/mg. The molecular weight and isoelectric point of the purified enzyme were 54,000 and pI 5.4, respectively. The optimal pH and temperature were 6.0 and $45^{\circ}C$, and the enzyme was stable between pH 6.5 and 7.5 and below $50^{\circ}C$. The estimated Km and Vmax were 10 mg/mL and $6.25{\mu}mol/min$ for carboxymethyl cellulose and 30.3 mg/mL and $2.85{\mu}mol/min$ for sigmacell, respectively. The enzyme was partially inhibited by $Ag^+$, $Zn^{+{+}}$, $Fe^{+{+}}$ and EDTA, while completely inhibited by $Cd^{+{+}}$ and $Hg^{+{+}}$ at 1 mM concentration.

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Purification and Characterization of Cellulolytic Enzymes from Aspergillus niger (Aspergillus niger가 생산(生産)하는 섬유소(纖維素) 분해효소(分解酵素)의 정제(精製) 및 특성(特性))

  • Park, Kwan-Hwa;Oh, Tae-Kwang;Shin, Jae-Doo
    • Applied Biological Chemistry
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    • v.24 no.3
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    • pp.186-193
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    • 1981
  • Three fractions of carboxymethyl-cellulase (F-I, F-II, and F-III) and ${\beta}-glucosidase$ form Aspergillus niger were partially purified by ammonium sulfate fractionation. Sephadex G-150 and DEAE-Sephadex column chromatography. The optimum conditions such as pH and temperature and thermal inactivation properties of the enzymes were investigated. Arrhenius plots of F-II and F-III appeared as straight lines, whereas that of F-I was biphasic. The Z-values of F-II and F-III were $8^{\circ}C$ and $10^{\circ}C$ respectively, while that of F-I was $4^{\circ}C$ over $60{\sim}70^{\circ}C$ and $383^{\circ}C$ over $70{\sim}98^{\circ}C$. Three fractions and the crude extract of carboxymethyl-cellulase exhibited a similar optimum pH 4.3 and temperature of $60^{\circ}C$, while Z-value of crude extract $(21.5^{\circ}C)$ was much higher than that of the purified enzyme. Maximum activity of both purified and crude extract of ${\beta}-glucosidase$ was shown at pH 4.7 and $60^{\circ}C$, and z-value of the enzyme was $7^{\circ}C$.

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Production of Lignocellulytic Enzymes from Spent Mushroom Compost of Pleurotus eryngii (큰느타리버섯 수확 후 배지로부터 리그닌섬유소분해효소 생산)

  • Lim, Sun-Hwa;Kim, Jong-Kun;Lee, Yun-Hae;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.40 no.3
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    • pp.152-158
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    • 2012
  • The lignocellulytic enzymes including a-amylase (EC 3.2.1.1), lignin peroxidase (EC 1.11.1.14), laccase (EC 1.10.3.2), xylanase (EC 3.2.1.8), ${\beta}$-xylosidase (EC 3.2.1.37), ${\beta}$-glucosidase (EC 3.2.1.21) and cellulase (EC 3.2.1.4) were extracted from spent mushroom compost (SMC) of Pleurotus eryngii. Different extraction buffers and conditions were tested for optimal recovery of the enzymes. The optimum extraction was shaking incubation (200 rpm) for 2 h at $4^{\circ}C$. ${\alpha}$-Amylase was extracted with the productivity range from 1.20 to 1.6 Unit/SMC g. Cellulase was recovered with the productivity range from 2.10 to 2.80 U/gf. ${\beta}$-glucosidase and ${\beta}$-xylosidase productivities showed lowest recovery producing 0.1 U/g and 0.02 U/g, respectively. The P. eryngii SMCs collected from three different mushroom farms showed different recovery on laccase and xylanse, cellulase. Furthermore, the water extracted SMC was compared to commercial enzymes for its industrial application in decolorization and cellulase activity.

Cloning of the Cellulase Gene and Characterization of the Enzyme from a Plant Growth Promoting Rhizobacterium, Bacillus licheniformis K11 (고추역병 방제능이 있는 식물성장촉진 균주 Bacillus licheniformis K11의 cellulase 유전자의 cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.50 no.2
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    • pp.95-100
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    • 2007
  • The cellulase gene of Bacillus licheniformis K11 which has plant growth-promoting activity by auxin and antagonistic ability by siderophore was cloned in pUC18 using PCR employing heterologous primers. The 1.6kb PCR fragment contained the full sequence of the cellulase gene, denoted celW which has been reported to encode a 499 amino acid protein. Similarity search in protein data base revealed that the cellulase from B. licheniformis K11 was more than 97% identical in amino acid sequence to those of various Bacillus spp. The cellulase protein from B. licheniformis K11, overproduced in E. coli DH5${\alpha}$ by the lac promoter on the vector, had apparent molecular weight of 55 kDa upon CMC-SDS-PAGE analysis. The protein not only had enzymatic activity toward carboxymethyl-cellulose (CMC), but also was able to degrade insoluble cellulose, such as Avicel and filter paper (Whatman$^{\circledR}$ No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. Consequently B. licheniformis K11 was able to suppress the peperblight causing P. capsici by its cellulase. Biochemical analysis showed that the enzyme had a maximum activity at 60$^{\circ}C$ and pH 6.0. Also, the enzyme activity was activated by Co$^{2+}$ of Mn$^{2+}$ but inhibited by Fe$^{3+}$ or Hg$^{2+}$. Moreover, enzyme activity was not inhibited by SDS or sodium azide.

Strain Improvement of Penicillium verruculosum for High Cellulase Production by Induced Mutation (섬유소분해효소 생산증진을 위한 Penicillium verruculosum의 균주개량)

  • 정기철
    • Microbiology and Biotechnology Letters
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    • v.15 no.6
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    • pp.388-395
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    • 1987
  • In order to obtain a regulatory mutant strain with high cellulase activity, a newly isolated Penicillium verrculosum, strain F-3 was used as parental strain since it was proved to be an efficient cellulase producer. A number of experiments were conducted to determine the optimum conditions to in-duce mutagenesis and isolate the desirable mutant strains. Out of several restriction compounds tested, 1.5% oxgall was found to be most effective to restrict the colony size by suppressing overgrowth. Derepression of catabolites was employed as a criterion in selecting mutant strains with high cellulase productivity. Production of cellulase by Penicillium venculosum F-3 was suppressed when cultured on the media with more than 1% of glucose or glycerol. It was found that either irradiation with UV light for 19 mins or treatment with nitrosoguanidine at 200$\mu\textrm{g}$/m1 for 60 mins, induced mutagenesis at desired level, when the survival rate of the spore was 0.2% and 48%, respectively. Three mutant strains of F-3, UV-9, UV-10, and NTG-3 that had the highest cellulase productivity were finally selected, based on filter paper degradation rate, size of clearing zone on the screening plate and cellulase activity in the medium containing cellulose powder. When the mutant strains were compared with parental strain F-3, on the KC-M-W medium containing cellulose powder, the filter paper activities of UV-9, UV-10, and NTG-3 were increased by 34%, 55%, and 41%, respectively. However, the assimilation of cellobiose octaacetate by UV-9 or NTG-3 was markedly reduced. When the mutant UV-10 was grown on cellobiose octaacetate medium (CCA-4) in shaking flasks, the cellulase activities of the mutant increased by 20 to 50% compared to the parental strain. Excreation of soluble protein from the mutant also elevated up to 30%. The mutant also constitutively produced both CMCase and $\beta$-glucosidase, though at relatively low level, in the presence of glucose or cellobiose as carbon sources.

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Conversion of Fermented Feed by Basidiomycetes (담자균(擔子菌)을 이용(利用)한 발효(醱酵) 사료(飼料)에 관한 연구(硏究))

  • Hong, Jae-Sik;Kim, Joong-Man;Jeong, Jin-Chul;Lee, Tae-Kyu;Kim, Dong-Han;Kim, Myung-Kon;Lee, Keug-Ro
    • The Korean Journal of Mycology
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    • v.13 no.3
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    • pp.157-168
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    • 1985
  • To investigate nutritive values of a feed fermented with basidiomycetes, among the isolated strains, Lyophyllum decastes (Fr.) Sing. was found with the greatest enzyme productivity and rapid mycelial growth in rice straw medium. Optimum temperature, pH and moisture content for mycelial growth and enzyme production of the strain were $25{\sim}30^{\circ}C,\;pH\;4.0{\sim}7.0\;and\;70{\sim}75\;%$, respectively. Fifteen days of culture were required for the highest enzyme productivity. Among the sub-materials added, $30{\sim}40\;%$ of rice bran and $10{\sim}20\;%$ of defatted perilla seeds were effective for the enzyme production, but caused a reduced mycelial growth. The greatest effect of an addition of inorganic salts was obtained with $0.36{\sim}0.72\;%\;of\;(NH_4)_2HPO_4$. When 40 mesh or smaller rice straw and steam treatment at $0.5\;kg/cm^2$ were used, the mycelial growth decreased, whereas the enzyme production increased. The mycelial growth and enzyme production increased when $Ca(OH)_2$ was used as the alkali treatment, but decreased with increasing concentration of NaOH. As the fermentation proceeded, the amounts of ash, reducing sugar and total nitrogen increased, but cellulose, lignin and pentosan decreased. When the rice straw was treated with alkali, the amounts of ash, total nitrogen and lignin decreased, but reducing sugar and cellulose increased. At higher NaOH concentration, the variation become greater. The in vitro dry matter digestibility of the products increased from 55.03 % at the beginning of the fermentation to 62.72 % at 45 days after fermentation. The most effective alkali treatment on the digestibility of rice straw was KOH followed by NaOH. However, the digestibility increased with increasing concentration of NaOH. The digestibility of pretreated with alkali increased after fermentation as well.

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