• Title/Summary/Keyword: 분해균주

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Characterization of Cellulases from Schizophyllum commune for Hydrolysis of Cellulosic Biomass (Schizophyllum commune에 의한 Cellulase 생산 및 섬유소계 바이오매스의 당화를 위한 효소적 특성)

  • Kim, Hyun-Jung;Kim, Yoon-Hee;Cho, Moon-Jung;Shin, Keum;Lee, Dong-Heub;Kim, Tae-Jong;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.6
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    • pp.547-560
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    • 2010
  • The optimum culture condition of Schizophyllum commune for the cellulase production and its enzymatic characteristics for saccharification of cellulosic biomass were analyzed. S. commune secrets ${\beta}$-1,4-xylosidase (BXL) and cellulases, including endo-${\beta}$-1,4-glucanase (EG), cellobiohydrolase (CBH), and ${\beta}$-glucosidase (BGL). The optimum reaction temperature for all cellulases was $50^{\circ}C$ and the thermostable range was $30{\sim}40^{\circ}C$C. The optimum reaction pH for all cellulases was 5.5 in a range of temperature from $0^{\circ}C$ to $55^{\circ}C$. The best nutritions for the cellulase production of S. commune among tested nutrients were 2% cellulose for the carbon source and corn steep liquor or peptone/yeast extract for the nitrogen source without vitamins. The environmental culture condition for the cellulase production was 5.5~6.0 for pH at $25{\sim}30^{\circ}C$. The enzyme activities of EG, BGL, CBH, and BXL were 3670.5, 631.9, 398.5, and 15.2 U/$m{\ell}$, respectively, after concentration forty times from the culture broth of S. commune which was grown at the optimized culture condition. Alternative filter paper unit assay showed 11 FPU/$m{\ell}$ enzyme activity. The saccharification tests using cellulase of S. commune showed the low saccharification rate on tested hardwoods but a high value of 50.5% on cellulose, respectively. The saccharification rate (50.5%) of cellulose by cellulase produced in this work is higher than 45.7% in the commercial enzyme (Celluclast 1.5L, 30 FPU/g, glucan).

The Whole Extract of Enterococcus faecalis Has Suppressive Effect on the Allergic Responses in Asthmatic Mouse Model (천식 마우스 모델의 알러지 반응에서 Enterococcus faecalis 전체 추출물의 억제 효과)

  • Chang, Jeong Hyun;Yang, EunJu;Yu, Sun Nyoung;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1168-1175
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    • 2017
  • Probiotics are usually defined as intestinal bacteria that provide healthy benefit to the host and may offer new therapeutic materials for the treatment of inflammatory diseases. Lactobacillus, Bifidobacterium and Enterococcus are known as typical probiotics. But, these bacteria have mostly a weak viability and thus decreased probiotics-mediated effects in the intestinal tract. Asthma is an inflammatory airway disease, which is characterized by the releases of inflammatory mediators including cytokine and IgE. They are mainly associated with the recruitment, activation and disregulation of specific inflammatory cells, especially mast cells, monocytes, T cells, eosinophils and neutrophils in asthma. We performed these studies as in vitro and in vivo test the human inflammatory cell lines and ovalbumin (OVA)-induced asthma mouse model. And then the inhibitory effects of Enterococcus faecalis whole extract on inflammatory responses were examined. For our examinations, the E. faecalis whole extract (Ef extract) was acquired from whole bacteria of E. faecalis using freeze/thawing after ultrasonication method. As results, OVA-mediated THP-1 cell viability was decreased by the treatment of Ef extract. In the asthmatic mouse model, Ef extract inhibited the infiltration of inflammatory cells into the inflammatory sites and blood. This whole extract may have anti-asthmatic effects associated with the regulation of IL-5 and IgE expression. It may also be a promising candidate in anti-allergic medicine for the treatment of asthma.

Quality characteristics and preparing of solid starter using fungal strains for Takju (탁주용 곰팡이 고체종국 제조 및 품질 특성)

  • Baek, Chang-Ho;Baek, Seong Yeol;Mun, Ji-Young;Choi, Han-Seok;Kang, Ji-Eun;Jung, Seok-Tae;Yeo, Soo-Hwan
    • Food Science and Preservation
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    • v.23 no.6
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    • pp.797-803
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    • 2016
  • In this study, we investigated the effect of fermentation conditions on the amylolytic and proteolytic activities of Aspergillus luchuensis strain 74-5 and Aspergillus oryzae strain 75-2, which are used in the preparation of the starter culture, for Takju (Korean traditional rice wine). The starter culture was optimized using different conditions, such as inoculum size, inoculation temperature, and incubation time. The enzyme activities under each condition were measured. In the A. luchuensis strain 74-5 starter culture, the ${\alpha}-amylase$ and glucoamylase activities increased, however the activity of acidic protease decreased as the diluent to starter culture ratio increased. In the A. oryzae 75-2 starter culture, all enzyme activities were maintained at a higher level even at 5% inoculation ratio. Higher enzyme activities were observed in the middle range of inoculation temperature (35, $40^{\circ}C$), than in the lower range (20, $30^{\circ}C$). Enzyme activity in the starter culture varied with incubation time, however it was the highest at 144 and 120 hr, respectively, for A. luchuensis strain 74-5 and A. oryzae strain 75-2. The spore count of the starter culture was approximately $2{\times}10^7$ during fermentation, out of which contamination by aerobic bacteria was about $3{\times}10^3$. The results suggested that the starter culture of each strain could be used as an inoculum for fermentation. However, we needs to conduct further research for the selection of suitable diluting agents as well as drying methods to reduce the contamination by aerobic bacteria, while retaining the enzyme activity.

Diversity, Saccharification Capacity, and Toxigenicity Analyses of Fungal Isolates in Nuruk (누룩곰팡이 분리균의 다양성 및 당화능 분석과 독소생산능 조사)

  • Kim, Min Sik;Kim, Sinil;Ha, Byeong-Seok;Park, Hye-Young;BaeK, Seong-Yeol;Yeo, Soo-Hwan;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.191-200
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    • 2014
  • Nuruk samples collected from various regions in Korea were investigated in terms of fungal contents and diversity. In measurement of colony forming unit (CFU) in Nuruk suspensions on DRBC agar, Nuruk samples MS4, MS8, and MS10 were among the highest fungal density, with $1,278.9{\pm}21.6$ (${\times}10^4$), $1,868.0{\pm}27.7$ (${\times}10^4$), and $775.1{\pm}19.2$ (${\times}10^4$) were among the samples showing the highest fungal density. CFU per 20 mg Nuruk, respectively. The majority of fungal components were yeasts, including Pichia anomala, P. kudriavzevii, Kluyveromyces marxianus, and Saccharomycopsis fibuligera, whereas Aspergillus oryzae and Rhizopus oryzae, the representative Nuruk fungi, were predominant only in the low fungal density Nuruks (MS2, MS5, and MS11). Saccharification capability of the fungal isolates was assessed by measurement of amylase activity in the culture broth. The highest amylase activity was found in A. niger and A. luchuensis, followed by S. fibuligera. A. oryzae and R. oryzae showed fair amylase activity but significantly lower than those of the three fungal species. R. oryzae was suggested to play an additional role in degradation of ${\beta}$-glucan in crop component of Nuruk since R. oryzae was the only fungus that showed ${\beta}$-glucanase activity among the fungal isolates. To confirm the safety of Nuruk, aflatoxigenicity of the isolated Aspergillus was estimated using the DNA markers norB-cypA, aflR, and omtA. All of the isolates turned out to be non-aflatoxigenic as evidenced by the deletion of gene markers, norB-cypA and aflR, and the absence of aflatoxin in the culture supernatants shown by TLC analysis.

Characterization and Purification of the Bacteriocin Produced by Bacillus licheniformis Isolated from Soybean Sauce (간장에서 분리한 Bacillus licheniformis가 생산하는 박테리오신의 특성 및 정제)

  • Jung, Sung-Sub;Choi, Jung-I;Joo, Woo-Hong;Suh, Hyun-Hyo;Na, Ae-Sil;Cho, Yong-Kweon;Moon, Ja-Young;Ha, Kwon-Chul;Paik, Do-Hyeon;Kang, Dae-Ook
    • Journal of Life Science
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    • v.19 no.7
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    • pp.994-1002
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    • 2009
  • A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.

Expression and Purification of Recombinant Human Interferon-gamma Produced by Escherichia coli (대장균이 생산한 재조합 인체 감마인터페론의 발현과 정제)

  • Park, Jung-Ryeol;Kim, Sung-Woo;Kim, Jae-Bum;Jung, Woo-Hyuk;Han, Myung-Wan;Jo, Young-Bae;Jung, Joon-Ki
    • KSBB Journal
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    • v.21 no.3
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    • pp.204-211
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    • 2006
  • For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

The Standing Crops and Soil-borne Microfungal Flora of Phyllostachys reticulata in Korea (한국산(韓國産) 왕대나무의 현존량(現存量)과 토양(土壤) 미세균류상(微細菌類相))

  • Kim, Kwan-Soo
    • The Korean Journal of Mycology
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    • v.7 no.2
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    • pp.91-116
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    • 1979
  • This paper is to investigate the standing crops and microfungal flora in soil in Phyllostachys reticulata forests in both the Yesan area (A) and the Kwangsan area (B). The stand density of the bamboo revealed 17,250 shoots per ha in area A, and in area B 14,780 shoots which were 16.1% less in number than area A. In respect to the environmental factors between the two areas, the mean temperature during the growth period was $1.5{\sim}2^{\circ}C$ higher in area B than in area A, soil tempeature also was $1{\sim}2^{\circ}C$ higher in area B, and the total quantities of nitrogen, phosphoric acid and organic compounds contained in the soil of area B were also slightly higher than those of area A. In area B the quantities of dried leaf matter, humus, and vegetation in the bamboo forest were also larger than in area A. In addition, five more species of microfungi which playa role in the decomposition of the various organic materials in the bamboo forests were identified in area B: Mortierella elongata, Mucor circinelloides, Aspergillus japonicus, Penicillium waksmani and Trichoderma lignorum. The atmospheric temperature in the inner portions of the bamboo forests was lower than the outside temperature, but the humidity was higher. The rates of relative illuminance were measured in area A at 4.19%, and in area B at 2.7%. These values revealed that the photosynthetic acitivity in the lower part of the bamboo was lost but it was considered that lower illuminance increased the microfungal activities in the vicinity of the surface soil. Since the productive structure of the bamboo showed that the maximum amount of photosynthesis was located in the upper portion of the bamboo in area B, it was considered to be an effective structure in maintaining the high productivity of the bamboo. The allometric relation between $D^2H$ and dry weight of stems(Ws), branches(Wb) and leaves(Wl) of the bamboo in area A were appoximated by log Ws=0.5262 log $D^2H$+1.9546; log Wb=0.6288 log $D^2H$+1.5723; log Wl=0.5181 log $D^2H$+1.8732, and those of the bamboo in area B were approximated by log Ws=0.5433 log $D^2H$+1.8610; log Wb=0.1630 log $D^2H$+2.3475; log Wl=0.4509 log $D^2H$+2.0041. From the above, the standing crops in area A were measured thus: Ws was 1,128. 83kg; Wb, 689.05kg; Wl, 926.69kg and Wl, 2,744.57kg per 10a. In area B, Ws was 1,206. 66kg; Wb, 679.92kg; Wl, 1,112.51kg and Wt, 2.999kg per l0a. Significant differences from the result of t-test were for $D^2H$ Ws, Wl and Wt between areas A and B. But no significant difference was found for Wb. In order to record as completely as possible the microfungal flora of the areas, every possible means was tried, and 158 strains of fungi were isolated, and of these, the microfungi of 55 species were identified. The dominant species were Trichoderma viride, Penicillium janthinellum, P. commune, Aspergillus oryzae, A. niger, A. gigantus, A. fumigatus, Mortierella ramaniana, var. anguliFPora, Mucor hiemalis and Zygorhynchus moelleri. According to the above results, it was revealed that optimum soil, the increases of soil materials, more species of soil microfungi, and the atmospheric temperature during the growth period have made the bamboo flourish and bring more species and larger quantities of vegetation in the bamboo forests. The correlation between the standing crops and environmental factors in the bamboo forest is considered to be a complicated relationship of all the factors, but the stand density is thought to be the most important factor involved.

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