• Title/Summary/Keyword: 단백질 동정

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Hydrolytic and Metabolic Capacities of Thermophilic Geobacillus Isolated from Litter Deposit of a Lakeshore (수변 낙엽퇴적층에서 분리한 호열성 Geobacillus의 물질 분해 특성)

  • Baek, Hyun-Ju;Zo, Young-Gun;Ahn, Tae-Seok
    • Korean Journal of Microbiology
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    • v.45 no.1
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    • pp.32-40
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    • 2009
  • To understand contribution of thermophilic microorganisms in decomposition of litter deposits on shore of lakes, we surveyed a lakeshore litter deposit for bacteria growing at $60^{\circ}C$. Ten thermophilic isolates were selected for in-depth characterization, based on their high capacity to degrade high molecular weight organic compounds. Based on phylogenetic analysis on their 16S rRNA gene sequences, all isolates were identified as Geobacillus. The optimal growth temperature and pH of the strains ranged $55{\sim}60^{\circ}C$ and 6.0${\sim}$8.0, respectively. Salinity was inhibitory to the growth of the isolates, showing marked decrease of growth rates at 3% salinity. Based on activities of hydrolytic enzymes and profiles of carbohydrate utilization (determined by API 50 CHB kit), three G. stearothermophilus strains showed patterns clearly distinctive from other isolates. Two G. kaustophilus strains also demonstrated distinctiveness in their metabolic pattern and ecological parameters. However, ecological and metabolic profiles of the other five isolates were more variable and showed some degree of digression from their phylogenetic classification. Therefore, it could be concluded that endospore-forming thermophilic bacteria in lakeshore litter deposits contribute to degradation of organic materials with diverse ecological niches while having successions similar to microbial flora in compost. We propose that the thermophilic isolates and/or their thermo-tolerant enzymes can be applied to industrial processes as appropriate mixtures.

Isolation and Characterization of Exopolysaccharide Producing Lactic Acid Bacteria from Kimchi (김치로부터 Exopolysaccharide 생성 유산균의 분리 및 특성 규명)

  • Kim Uyo-Ju;Chang Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.196-203
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    • 2006
  • Three slime-forming lactic acid bacteria were isolated from Kimchi and shown to produce viscous exopolysaccharides (EPS) in sucrose media. The isolated strains, GJ2, C3 and C11, were identified as Leuconostoc kimchii, Leuconostoc citreum and Leuconostoc mesenteroides, respectively, by examining their metabolic characteristics and determining their 16S rDNA sequences. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 exhibited high viability (maintained initial viable cell count of $10^8$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastric juice for 2 h and in 0.3% oxgall for 24 h. When tested, Leu. kimchii GJ2, in particular, displayed antimicrobial activity against pathogenic microorganisms. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 produced 21.49 g/l, 16.46 g/l and 22.98 g/l EPS, respectively, in sucrose (5%) medium. The amount of purified EPS extracted from Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 was 14.61 g/l, 7.73 g/l and 4.77 g/l, respectively. Although the EPS produced by Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 differed in viscosity, TLC and HPLC analysis revealed that each contained only one type of monosaccharide, glucose. The average molecular mass of EPS produced by Leu. kimchii GJ2 was 306,606 Da.

Exploration of optimal Lactobacillus plantarum strains for curdling milk for yogurt and evaluation of physicochemical and sensory properties (호상 요구르트에 적합한 Lactobacillus plantarum strains 탐색 및 요구르트의 이화학적 및 관능 특성)

  • Jeong, Seong-Yeop;Lee, Yong Hyen;Kang, Suna;Shin, Bae Keun;Park, Sunmin
    • Korean Journal of Food Science and Technology
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    • v.48 no.6
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    • pp.548-554
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    • 2016
  • Since some strains of kimchi lactobacilli can curdle milk, they can be used for making yogurt. However, the best Lactobacillus plantarum strains for curdling milk for yogurt are still unknown. In this study, we determined the best L. plantarum strains for curdling milk, and the physicochemical properties of yogurts made using different L. plantarum strains were examined. Three strains of L. plantarum useful for curdling milk were identified (YD2, YD9, YD12). The number of lactobacilli was lower in yogurts made with L. plantarum than in those made with control, and among the L. plantarum strains tested, YD12 had the highest bacterial counts. However, the microbial count reached $6.3{\times}10^8CFU/mL$ after 24-h fermentation in all yogurts. The pH of the yogurts decreased after 12-h fermentation, while the acidity increased. The low pH and high acidity decreased the viscosity in all the three types of yogurts, because the acids disturbed gel formation due to protein denaturation. Sensory evaluation revealed that the YD12 group showed a high percentage of completion similar to the control group. YD2 and YD9 showed a high sourness value and low sweetness value, whereas YD12 yielded optimal values for all the organoleptic characteristics. Therefore, YD12 would be a high quality bacterial strain for use as a yogurt starter culture.

Roles of Prostatic Acid Phosphatase in Prostate Cancer (Prostatic acid phosphatase의 전립선 암에서의 역할)

  • Kong, Hoon-Young;Lee, Hak-Jong;Byun, Jong-Hoe
    • Journal of Life Science
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    • v.21 no.6
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    • pp.893-900
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    • 2011
  • Prostatic acid phosphatase (PAP) is one of the widely used biomarkers in the diagnosis of prostate cancer. It was initially identified in 1935 and is the most abundant phosphatase in the human prostate. PAP is a prostate-specific enzyme that is synthesized in prostate epithelial cells. It belongs to the acid phosphatase group that shows enzymatic activity in acidic conditions. PAP is abundant in prostatic fluid and is thought to have a role in fertilization and oligospermia. It also has a potential role in reducing chronic pain. But one of the most apparent functions of PAP is the dephosphorylation of macromolecules such as HER-2 and PI3P that are involved in the ERK1/2 and MAPK pathways, which in turn leads to inhibition of cell growth and tumorigenesis. Currently, clinical trials using PAP DNA vaccine are underway and FDA-approved immunotherapy using PAP is commercially available. Despite these clinically important aspects, molecular mechanisms underlying PAP regulation are not fully understood. The promoter region of PAP was reported to be regulated by NF-${\kappa}B$, TNF-${\alpha}$, IL-1, androgen and androgen receptors. Here, the features of PAP gene and protein structures together with the function, regulation and roles of PAP in prostate cancer are discussed.

Composition of a Medium for Serum-free Culture of an Adipose-derived Stem Cell Line Established with a Simian Virus 40 T Antigen (Simian virus 40의 T항원 도입으로 수립한 지방유래줄기세포주의 효율적인 무혈청 배양법 및 무혈청 배지조성)

  • Kim, Gyu Bin;Joo, Woo Hong;Kim, Dong Wan
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1301-1307
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    • 2014
  • Adipose-derived stem cells (ADSCs) are considered promising tools for tissue regeneration. However, ADSCs have very poor proliferation capacity. Therefore, fetal bovine serum (FBS) is generally added to the culture media of ADSCs. As FBS contains many uncharacterized components that may affect cellular functions, methods for serum-free cultures of ADSCs have been widely investigated. In this study, to develop an efficient method for a serum-free culture of ADSC-T, we used an ADSC line established by introducing the simian virus 40 (SV40) T gene into primary ADSCs. We then investigated the effect of amino acids, vitamins, and other components on the growth of ADSC-T. When the ADSC-T cells were plated with DMEM/F12 serum-free medium, the cells did not proliferate, and the mixture of amino acids, vitamins, and B27 supplement did not increase the growth of the cells. However, when the ADSC-T cells were provided with serum-free DMEM/F12 after they had been cultured with serum-supplemented DMEM for 24 h, the cells proliferated, and the vitamins and B27 supplement increased the cell growth. Stem-Pro serum-free medium also appeared to be useful as a suspension culture for the ADSC-T cells. The ADSC-T cells secreted large amounts of proteins of around 70 kDa. Insulin-like growth factor (IGF) and fibroblast growth factor basic (FGF basic) were secreted by ADSC-T in larger amounts in the serum-free culture than in the serum-supplemented culture.

Identification of Substrate Specificity Determinant of Achromobacter Protease I (API) and Catalytic Activity of Mutant D225E for Ornithine Substrate (Achromobacter Protease I (API)의 기질특이성 결정기의 동정과 변이체[D225E]의 Ornithine 기질에 대한 촉매활성)

  • Lim, Seong-Il;Kwon, Oh-Jin;Choi, Cheong
    • Applied Biological Chemistry
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    • v.40 no.3
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    • pp.189-195
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    • 1997
  • The structural basis of Iysine specificity of Achromobacter protease I (API) was investigated by means of site-directed mutagenesis. The precursor protein in which Glu190, one of the two candidates for determining Iysine specificity, was substituted by glutamine, aspartic acid or leucine was processed autocatalytically to attaln full pretense activity with lysine specificity. The substitution of the other candidate, Asp225, for asparagine or leucine produced no mature active forms of pro-API. The precursor protein of the mutant D225E slowly matured autocatalytically. The lysylendopeptidase activity of the mature D225E was 0.25% of that of native API, and this reduced activity is mainly due to a decrease in the affinity of the enzyme for lysine. These results suggest that Asp225 plays a critical rol in restricted substrate specificity as a lysylendopeptidase. However, D225E exhibited no measurable activity for synthetic ornithine substrate. Since the hydroxyl group of Ser194 in this mutant retained essentially the same reactivity to DFP or PMSF as that in native API, it can be noted that a methylene unit longer side chain of residue 225 is not compensated by a methylene unit shorter side chain at subsite P1 in the bound substrate.

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Isolation and Characterization of the nsdC Gene in Sexual Development of Aspergillus nidulans (Aspergillus nidulans의 유성분화에 관여하는 nsdC 유전자의 분리 및 분석)

  • Kim, Hye-Ryun;Han, Dong-Min
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.246-251
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    • 2006
  • A lot of mutants which cannot initiate sexual development were screened and several loci including nsdA, nsdB, nsdC, and nsdD were identified in homothallic ascomycetes Aspergillus nidulans. The NSD206, which has nsdC6 allele, showed typical phenotype of NSD (Never in sexual development) mutants. The nsdC gene was cloned by transforming NSDP697 ($nsdC^-$, $pryG^-$) with AMA1-NotI genomic library. The transforming library DNA recovered from several transformants showing wild phenotype carried about 10 kb genomic DNA insert. The DNA sequence of nsdC was analysed using GPS (Genome priming system). The nsdC gene has an open reading frame (ORF) of 1,929 bp encoding a putative polypeptide of 643 amino acids. The NsdC carries $C_2H_2C_2H_2C_2HC$ type zinc finger DNA binding domains in the middle of the polypeptide. A coiled-coil domain at its C terminus were also found. In nsdC6 allele, a single T insertion was occurred between 407-408 bp leading to the frameshift mutation and early termination of translation producing the truncated protein which has only 139 amino acids.

Growth Inhibition of Human Hepatoma and Bladder Carcinoma Cells by DNA Topoisomerae Inhibitor β-lapachone (DNA topoisomerase 억제제인 β-lapachone에 의한 인체 간암 및 방광암세포 증식억제에 관한 연구)

  • Choi Da Yean;Lee Jae Il;Chung Hyun Sup;Seo Han Gyeol;Woo Hyun Joo;Choi Yung Hyun
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.323-331
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    • 2005
  • The objective of the present study was to investigate the effect of $\beta-lapachone$, a quinone obtained from the bark of the lapacho tree (Tabebuia avellanedae) in South America, on the cell growth of human hepatoma (HepG2) and bladder (T24) carcinoma cells. Exposure of cancer cells to $\beta-lapachone$ resulted in growth inhibition, morphological changes and apoptosis in a concentration-dependent manner, which could be proved by MTT assay and flow cytometry analysis. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyses revealed that $\beta-lapachone$ did not affect the levels of tumor suppressor p53 and cyclin-dependent kinase (Cdk) inhibitor p21 (WAFl/CIPl) expression. However, the transcriptional factor Sp-l and proliferating cell nuclear antigen (PCNA) protein levels were significantly down-regulated by $\beta-lapachone$ in both cell lines. Moreover, $\beta-lapachone$ treatment caused a dose-dependent inhibition of the expression of telomere regulatory gene products such as human telomere reverse transcriptase (hTERT) and telomerase-associated protein-l (TEP-l). Taken together, these findings suggest that $\beta-lapachone$-induced inhibition of human hepatoma and bladder carcinoma cell proliferation is associated with the induction of apoptotic cell death via modulation of several major growth regulatory gene products, and provide important new insights into the additional mechanisms of the anti-cancer activity of $\beta-lapachone$.

Fractionation and Physicochemical Characteristics of Caprine Casein by DEAE-Cellulose (DEAE-Cellulose에 의한 산양유 Casein의 분별 및 이화학적 성질에 관한 연구)

  • Im, Dong-Hyeon;Jeon, U-Min;Han, Gyeong-Sik;Kim, Byeong-Cheol;Hwang, Gwang-Yeon;Kim, Se-Heon
    • Journal of Dairy Science and Biotechnology
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    • v.18 no.1
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    • pp.1-8
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    • 2000
  • This experiment was carried out to study fractionation and physicochemical characteristics of caprine casein. Acid caseins obtained from caprine colostral and normal milk were analyzed by chymosin treatment and fractionated by DEAE-cellulose column chromatography with linear gradient and electrophoresis. Protein, fat and lactose of caprine normal milk were 2.70${\pm}$0.27%, 3.82${\pm}$0.51%, and 4.10${\pm}$0.29%, respectively. More non-protein nitrogen(NPN) was released by chymosin treatment from caprine colostral casein than normal casein. The electrophoretic pattern of caprine casein was not similar to that of bovine casein, Caprine normal casein was fractionated by DEAE-cellulose column chromatography with a 0.08${\sim}$0.18 M NaCl linear gradient into 5 pes with the proportion of 5.27%, 26.07%, 25.97%, 30.40% and 12.29%, respectively. In order to identify the pure fraction, the chymosin-treated caprine normal casein was fractionated by DEAE-cellulosecolumn chromatography with a 0.08${\sim}$0.18 M NaCl linear gradient into 6 peaks with the proportion of 17.06%, 9.10%, 17.85%, 20.11%, 27.03% and 8.85%, respectively.

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A Secure Access Control Model for Privacy Protection using Purpose Classification (사용목적 분류화를 통한 프라이버시 보호를 위한 보안 접근제어 모델)

  • Na Seok-Hyun;Park Seog
    • Proceedings of the Korean Information Science Society Conference
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    • 2006.06c
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    • pp.265-267
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    • 2006
  • 사용목적(Purpose)은 최근 개인 프라이버시 보호와 관련하여 데이타 수집과 수집 후 보안관리에 있어서 중요한 요소로 사용되고 있다. W3C(World Wide Web Consortium)는 데이타 제공자가 자신이 방문한 웹 사이트에 개인정보를 제공하는 것을 통제할 수 있도록 하는 표준을 제시하였다. 그러나 데이타 수집 후 유통과정에서 개인정보에 대한 보안관리에 대한 언급이 없다. 현재 히포크라테스 데이타베이스(Hippocratic Databases), 사용목적기반 접근제어(Purpose Based Access Control)등은 W3C의 데이타 수집 메커니즘을 따르고 있으며, 데이타 수집 후 보안관리에 대하여 사용목적 관리와 접근제어 기법을 사용하여 관리를 하고 있으나 사용목적에 대한 표현과 사용목적 관리의 미흡함으로 인하여 그에 따르는 개인정보의 프라이버시 보호에 있어서 효과적인 해결책을 제시하지 못하고 있다. 본 논문은 사용목적의 표현력을 향상시키면서. 사용목적의 효과적인 관리기법을 제시한다. 또한 개인의 프라이버시 보호를 위한 방법으로 사용목적의 분류화를 통해 최소권한의 원칙을 따르는 접근제어 기법을 제시한다. 본 논문에서는 사용목적을 상속적, 시간적 그리고 독립적 구조로 분류화하였으며, 이렇게 분류화된 사용목적에 대한 각기 다른 관리기법을 제시한다. 또한 접근제어의 유연성을 위해 RBAC의 역할계층 구조를 사용하였으며, 일의 최소 단위인 태스크(task)의 최소권한을 얻기 위한 조건으로 몇몇 특성의 사용목적을 사용하여 만족할 경우 태스크를 처리하기 위한 기존 모델보다 향상된 최소사용권한을 제공하는 기법을 제시한다. Interference Contrast)에 의한 내부구조 관찰이 최종 동정기준이 되어야할 것으로 나타났다.cillus로 구성되었다. 한편, DAL세균군(42균주)은 high G+C 및 low G+C gram positive 계통군 이외에도 proteobacteria -subdivision에 속하는 Afipia와 Ralstonia, proteobacteria -subdivision에 속하는 Variovorax, proteobacteria $\beta$-subdivision에 속하는Pseudomonas로 구성되어 계통학적으로 다양한 세균임이 확인되었다. 40%까지 대체가 가능하였으며, 아울러 높은 라이신 부산물의 대체 수준에 있어서 사료효율과 단백질 전환효율을 고려한다면 아미노산 첨가(라이신과 아르지닌)와 중화 효과에 좋은 결과가 있을 것으로 사료된다.의한 적정 양성수용밀도는 각고 5~6cm 크기의 경우 10~15개체가 적합하였다. 수증별 성장은 15~20 m 수층에서 빨랐으며, 성장촉진과 폐사를 줄이기 위해서는 고수온이 지속되는 7~10월에는 20~30m수층으로 채롱을 내려 양성하고 그 외 시기에는 15 m층 내외가 좋은 것으로 나타났다. 상품으로 출하 가능한 크기 인 각고 10 cm이상, 전중량 140 g 내외로 성장시 키기까지는 채묘후 22개월이 소요되었고, 출하시기는 전중량 증가가 최대에 이르는 3월에서 4월 중순이 경제적일 것으로 판단된다.er 90 % of good relative dynamic modulus of elasticity due to fineness of formation caused by the

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