• 제목/요약/키워드: culture supernatant

검색결과 595건 처리시간 0.024초

효모에서 활성형의 곤충유래 항균펩티드 defensin의 발현 (Expression of Biologically Active Insect-Derived Antibacterial Peptide, Defensin, in Yeast)

  • 강대욱;안순철;김민수;안종석
    • 생명과학회지
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    • 제12권4호
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    • pp.477-482
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    • 2002
  • 효모 glucoamylase의 promoter와 분비신호서열 그리고 MF$\alpha$1의 prosequence를 이용하여 곤충 defensin을 S. cerevisiae 2805에서 항균활성을 보유한 형태로 발현 및 분비하는데 성공하였다. 발현된 defensin의 대부분이 세포 외로 분비되어 거의 모든 항균활성이 배양 상등액에 존재하였다. 이것은 S. cerevisiae에서 발현된 defensin이 glucoamylase의 분비신호서열과 MF$\alpha$1의 prosequenre에 의해 효율적으로 processing되어 분비됨을 시사한다. Defensin의 다른 미생물에 대한 항균활성을 조사한 결과 병원균인 St. aureus와 L. monocytogenes에 대해서도 항균활성이 존재하였다.

High-yield Purification and Characterization of Recombinant Human Leukotactin-1 in Pichia pastoris

  • Lim, In-Hwan;Lee, Kong-Ju;Lee, Eun-Kyoung;Park, Mu-Rim;Lee, Gue-Wha;Yeup Yoon;Park, Doo-Hong;Jung, Kyung-Hwan
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권1호
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    • pp.1-6
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    • 2004
  • The human chemokine, the short version of leukotactin-1(shLkn-1;molecular weight=7.2 kD and 66 amino acids), was expressed and secreted into a culture medium using the me-thylotrophic yeast, Pichia pastoris. The recombinant shLkn-1 was purified from the culture supernatant using a simple two-step procedure consisting of cation exchange and reverse phase chromatography(RPC), in which shLkn-1 was highly purified (99.5%) with a high recovery yield of 82.7%. The C-terminal truncated derivative of shLkn-1 was found in the supernatant and was separated by RPC. The physicochemical properties of the purified shLkn-1 were verified to be the same as expected. The biological activity of the purified recombinant shLkn-1 was also quantified using a chemotaxis assay. It was observed that the recombinant shLkn-1 had the maximum migration activity at a concentration of 10nM, as potent as MIP-1${\alpha}$.

구취를 유발하는 혐기성 세균의 증식을 억제하는 유산 간균의 분리 및 동정 (Isolation and identification of Lactobacillus inhibiting the production of halitosis by anaerobic bacteria)

  • 김미형;김선미
    • 한국치위생학회지
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    • 제4권2호
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    • pp.153-163
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    • 2004
  • There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic acid bacteria were isolated from normal inhabitants of children 's oral cavity, which inhibited the the production of halitosis by anaerobic bacteria. The authors identified the isolates by the lest using API 50 CHL medium kit. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. The optical density was 1.286 in the supernatant of Fusobacterium nucleatum after vortexing for 30 minutes, whereas in the supernatant of combined Fusobacterium nucleatum and each isolate, they were reduced to 0.628 and 0.497, which the percentages of coaggregation between them were 29.4% and 57.8%, respectively. 3. The optical density of Fusobacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$, being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusobacterium nucleatum and each isolate. 4. The optical density of Porphyromonas gingivalis precipitate was 1.932 in the culture media, being reduced to 1.170 and 1.266 in the coaggregated precipitates of Porphyromonas gingivalis and each isolate. 5. When two isolates were tested with API 50 CHL medium kit, those were identified as Lactobaciallius salivarius and Lactobacillus delbrueckii subsp. lactis.

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김치로부터 항돌연변이 물질을 생산하는 유산균의 분리 및 특성 (Isolation and Characterization of Lactic Acid Bacteria Producing Antimutagenic Substance from Korean Kimchi)

  • 이창호;박희동
    • 한국미생물·생명공학회지
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    • 제27권1호
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    • pp.15-22
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    • 1999
  • Various lactic acid bacteria were isolated from Korean Kimchi in order to study their antimutagenic substances. Ames test using Salmonella typhimurium TA98 and TA100 showed the strain KLAB21 to have the highest antimutagenic activity among the 230 isolated strains against MNNG (N-methyl-N'-nitro-N-nitrosoguanidine), NPD (4-nitro-O-phenylenediamine), NQO (4-nitrosoquinoline-1-oxide) and AFB1 (aflatoxin B1). The strain was identified as Lactobacillus plantarum based on its morphological, cultural and physiological characteristics. Antimutagenic activity of L. plantarum KLAB21 was found in culture supernatant suggesting the bacterium secrete antimutagenic substance in the media. No mutagenic activity was found in the culture supernatant. The isolated strain L. plantarum KLAB21 showed much higher antimutagenic activity than L. plantarum IAM1261 which is being used industrially for fermented milk production. The antimutagenic activity of L.plantarum KLAB21 was reconfirmed by the spore-rec assay using spores of Bacillus subtilis H17($Rec^+$) and M45($Rec^-$).

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Antagonism against Helicobacter Pylori and Proteolysis of Lactobacillus Helveticus CU631 and Strain Identification

  • Yoon, Y.H.;Won, B.R.
    • Asian-Australasian Journal of Animal Sciences
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    • 제15권7호
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    • pp.1057-1065
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    • 2002
  • The antagonistic activities of 30 strains of lactobacilli against Helicobacter pylori were determined and Lactobacillus helveticus CU631 has been selected as the strain which possesses the strongest inhibitory effect in the disc diffusion assay showing inhibition zone diameter of $10{\pm}1.5mm$, whereas those of L. plantarum and L. fermentum have been shown to be $4.0{\pm}0.6mm$. H. pylori G88016 revealed the highest vacuolating toxin producing activity among the 8 strains, the inhibitory activity of L. helveticus CU631 in vacuolating toxin producing activity of H. pylori manifested in the co-culture of two strains and in the 5:5 mixture of supernatant of the two strains. Both L. helveticus CU631 and cell free culture supernatant had a strong inhibitory activities in urease and cytotoxin producing activities of H. pylori NCTC11637 and CJH12. An accelerated proteolytic activity of water soluble peptides by L. helveticus CU631 during the refrigeration storage has been manifested in the cream cheese. DNA seqences of 16S-23S ribosomal RNA spacer region showed typical pattern among the various strains of L. helveticus, which could be used in the identification of L. helveticus CU 631.

Enhanced reutilization value of shrimp-shell waste via fed-batch biodegradation with higher production of reducing sugar, antioxidant, and DNA protective compounds

  • Rashid, Harun Ar;Jung, Hyun Yi;Kim, Joong Kyun
    • Fisheries and Aquatic Sciences
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    • 제21권10호
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    • pp.33.1-33.11
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    • 2018
  • As a process for commercial application, production of reducing sugar, antioxidant, and DNA protective compounds from shrimp-shell powder was investigated in a fed-batch biodegradation using Bacillus cereus EW5. The fed-batch biodegradation was operated in a 5-L bioreactor for 96 h according to three times pulse-feeding strategy. On the basis of the equal working volume (3 L), the fed-batch biodegradation showed a better production of the target compounds than the batch biodegradation, with higher cell density and shortened biodegradation period. The maximum values of the target compounds were 0.297 mg/mL of reducing sugar, 92.35% DPPH radical scavenging activity, 98.16% ABTS radical scavenging activity, and 1.55 reducing power at $A_{700}$, which were approximately 12.1, 3.4, 5.2, and 8.4% enhanced, respectively, compared with those obtained from the batch biodegradation. The fed-batch culture supernatant also showed the enhanced DNA damage inhibition activity than the batch culture supernatant. As a result, the fed-batch biodegradation accompanied by high cell density could produce more useful compounds, enabling an increase in the reutilization value of shrimp-shell waste.

Molecular Cloning and Analysis of Sporulation-Specific Glucoamylase (SGA) Gene of Saccharomyces diastaticus

  • Kang, Dae-Ook;Hwang, In-Kyu;Oh, Won-Keun;Lee, Hyun-Sun;Ahn, Soon-Cheol;Kim, Bo-Yeon;Mheen, Tae-Ick;Ahn, Jong-Seog
    • Journal of Microbiology
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    • 제37권1호
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    • pp.35-40
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    • 1999
  • Sporulation-specific glucoamylase (SGA) gene was isolated from genomic library of Saccharomyces diastaticus 5114-9A by using glucoamylase non-producing mutant of S. diastaticus as a recipient. When the glucoamylase activities of culture supernatant, periplasmic, and intracellular fraction of cells transformed with hybrid plasmid containing SGA gene were measured, the highest activity was detected in culture supernatant. SGA produced by transformant and extracellular glucoamylase produced by S. diastaticus 5114-9A differed in enzyme characteristics such as optimum temperature, thermostability, and resistance to SDS and urea. But the characteristics of SGA produced by sporulating yeast cells and vegetatively growing transformants were identical.

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Immune Enhancing Effect by Orally-Administered Mixture of Saccharomyces cerevisiae and Fermented Rice Bran

  • KOH, JONG HO;JIN MAN KIM;HYUNG JOO SUH
    • Journal of Microbiology and Biotechnology
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    • 제13권2호
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    • pp.196-201
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    • 2003
  • The mixture (PM) of Saccharomyces cerevisiae and fermented rice bran on the activation of macrophage and bone marrow cell proliferation was studied in mice. PM stimulated not only the activation of macrophage (1.8-fold of saline) but also IL-6 production from macrophage (1.5-fold) at 2.0 g/㎏/day during 7 days of oral administration. By the culture supernatant of Peyer's patch cells from C3H/HeJ mice fed PM at 2.0 g/㎏/day for 7 days, the bone marrow cells significantly proliferated compared with that of mice receiving only saline (1.7-fold). In addition, the contents of GM-CSF and IL-6 in the culture supernatant of Peyer's patch cells from mice fed PM at 2.0 g/㎏/day were increased in comparison with those from the control (1.8 and 1.4-fold, respectively). These results revealed that oral administration of PM may modulate IL-6 production to induce the activation of macrophage, and also enhance secretion of hematopoietic growth factors such as GM-CSF and IL-6 from Peyer's patch cells.

A Mouse Thymic Stromal Cell Line Producing Macrophage-Colony Stimulating Factor and Interleukin-6

  • Lee, Chong-Kil;Kim, Jeong-Ki;Kim, Kyungjae;Han, Seong-Sun
    • Archives of Pharmacal Research
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    • 제23권3호
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    • pp.252-256
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    • 2000
  • A thymic stromal cell line, TFGD, was established from a thymic tumor mass developed spontaneously in p53 knock out mouse, and was found to produce cytokines that could induce bone marrow hematopoietic stem cells (HSCs) to differentiate into macrophages. The cytokines produced by the TFGD line were assessed by immunoassays. High level of macrophage-colony stimulating factor (M-CSF) and interleukin (IL)-6 was detected in the TFGD-culture supernatant, whereas granulocyte/macrophage-colony stimulating factor (GM-CSF), IL-3, IL-4, IL-5, IL-13, or interferon (IFN)-$\gamma$ was undetectable. Blocking experiments showed that anti-M-CSF monoclonal antibody could neutralize the differentiation-inducing activity shown by the TFGD-culture supernatant. Dot blot analysis of the total RNA isolated from the cultured fetal thymic stromal cells showed that M-CSF transcripts were expressed in the normal thymus. These observations, together with the earlier finding that M-CSF plus IL-6 is the optimal combination of cytokines for the induction of macrophage differentiation from HSCs in vitro, may indicate that thymic macrophages could be generated within the thymus by cytokines involving M-CSF.

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세균 배양액과 반응한 Lactobacillus plantarum의 생장속도 및 항균 활성에 대한 연구 (Growth Rate and Antimicrobial Activity of Lactobacillus plantarum Reacting with Bacterial Culture Supernatant)

  • 하은미
    • 한국미생물·생명공학회지
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    • 제43권3호
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    • pp.187-194
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    • 2015
  • 인간의 위장관은 매우 복잡한 장내균총으로 이루어져 있으며, 장내균총은 인간의 생리 기능에 있어 매우 중요한 역할을 담당하고 있다. 특히 유아기의 장내균총의 불균형은 면역연관 질병을 일으키는 요인이며 치료 목적으로 probiotics가 유용하게 활용되고 있다. Probiotics의 대표 균종은 L. plantarum으로서 영유아기에 병원균보다 가장 먼저 정착해야 면역 질환을 예방할 수 있다. 그러나 아직 Lactobacillus가 장의 우점종으로 정착하기 위한 환경적 요인과 항생물질분비 조절에 대한 이해는 부족한 실정이다.