• 제목/요약/키워드: substrate resistance

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Identification and Characterization of Lactobacillus salivarius subsp. salivarius from Korean Feces

  • Bae, Hyoung-Churl
    • 한국축산식품학회:학술대회논문집
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    • 한국축산식품학회 2004년도 정기총회 및 제33차 춘계 학술대회
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    • pp.89-119
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    • 2004
  • This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

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C57BL/Ksj-db/db 제 2형 당뇨모델을 이용한 갈색거저리 유충(밀웜) 추출물의 인슐린 감수성 및 혈당개선효과 (Tenebrio molitor (Mealworm) Extract Improves Insulin Sensitivity and Alleviates Hyperglycemia in C57BL/Ksj-db/db Mice)

  • 김선영;박재은;한지숙
    • 생명과학회지
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    • 제29권5호
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    • pp.570-579
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    • 2019
  • 당뇨병은 서구화된 식습관으로 발생하는 심각한 만성대사질환의 하나이며, 당뇨병의 치료는 혈당을 정상적인 수준으로 유지하며 당뇨 합병증을 예방하는 것이다. 따라서 본 연구는 당뇨병 및 인슐린 저항성에 대한 유전적 분석에 널리 이용되는 C57BL/Ksj-db/db 당뇨동물모델을 이용하여 갈색거저리 유충(밀웜) 추출물의 6주간 섭취가 혈당개선에 미치는 영향에 대해 조사하고 이에 인슐린 민감성 개선과 당대사 조절을 통한 항당뇨 효과를 규명하고자 하였다. 제 2형 당뇨동물모델 실험 결과, db/db-MWE군(식이 0.5%)이 db/db-control군에 비해 유의적(p<0.05)으로 혈당이 감소하였다. 약물군인 db/db-RG군(식이 0.05%)은 부작용에 의해 눈에 띄게 체중이 증가하였으나, db/db-MWE군에서는 약물군에서의 체중증가와 같은 큰 부작용 없이 혈당 감소효과를 나타내었다. HbA1c와 혈장인슐린 농도의 경우 db/db-control군에 비해 db/db-MWE군이 유의적(p<0.05)으로 낮았다. 또한 골격근에서 p-IRS, p-AKT, PM-GLUT4의 발현을 확인한 결과, db/db-MWE군에서 db/db-control군에 비해 p-IRS, p-AKT, PM-GLUT4의 발현이 증가된 것을 알 수 있었다. 이는 밀웜 추출물의 섭취가 골격근 내로 당이 원활이 유입되도록 도와주어 인슐린 민감성을 개선시키며, 고혈당 증상을 개선시킨 것으로 사료된다. 밀웜 추출물을 식이에 0.5% 첨가하여 6주간 C57BL/Ksj-db/db 당뇨동물모델에 제공한 결과, 공복혈당과 HbA1c의 감소 및 인슐린 저항성을 개선시켰다. 이는 인슐린 민감성을 증가시키고, 당 대사 조절을 통해 고혈당 증상의 완화에 기인한 것으로 보인다. 따라서 밀웜은 당뇨병의 예방과 치료에 유용한 소재가 될 것으로 기대되며, 향후 제 2형 당뇨병 개선을 위해 더욱 다양한 연구가 이루어져야 할 것으로 사료된다.

고전압 전력반도체 소자 개발을 위한 단위공정에서 식각공정과 이온주입공정의 영향 분석 (Analysis of the Effect of the Etching Process and Ion Injection Process in the Unit Process for the Development of High Voltage Power Semiconductor Devices)

  • 최규철;김경범;김봉환;김종민;장상목
    • 청정기술
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    • 제29권4호
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    • pp.255-261
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    • 2023
  • 파워반도체는 전력의 변환, 변압, 분배 및 전력제어 등을 감당하는데 사용되는 반도체이다. 최근 세계적으로 고전압 파워반도체의 수요는 다양한 산업분야에 걸쳐 증가하고 있는 추세이며 해당 산업에서는 고전압 IGBT 부품의 최적화 연구가 절실한 상황이다. 고전압 IGBT개발을 위해서 wafer의 저항값 설정과 주요 단위공정의 최적화가 완성칩의 전기적특성에 큰 변수가 되며 높은 항복전압(breakdown voltage) 지지를 위한 공정 및 최적화 기술 확보가 중요하다. 식각공정은 포토리소그래피공정에서 마스크회로의 패턴을 wafer에 옮기고, 감광막의 하부에 있는 불필요한부분을 제거하는 공정이고, 이온주입공정은 반도체의 제조공정 중 열확산기술과 더불어 웨이퍼 기판내부로 불순물을 주입하여 일정한 전도성을 갖게 하는 과정이다. 본 연구에서는 IGBT의 3.3 kV 항복전압을 지지하는 ring 구조형성의 중요한 공정인 field ring 식각실험에서 건식식각과 습식식각을 조절해 4가지 조건으로 나누어 분석하고 항복전압확보를 위한 안정적인 바디junction 깊이형성을 최적화하기 위하여 TEG 설계를 기초로 field ring 이온주입공정을 4가지 조건으로 나누어 분석한 결과 식각공정에서 습식 식각 1스텝 방식이 공정 및 작업 효율성 측면에서 유리하며 링패턴 이온주입조건은 도핑농도 9.0E13과 에너지 120 keV로, p-이온주입 조건은 도핑농도 6.5E13과 에너지 80 keV로, p+ 이온주입 조건은 도핑농도 3.0E15와 에너지 160 keV로 최적화할 수 있었다.

Acyclovir저항성 Herpes Simplex Virus의 복제, DNA합성 및 형질 발현에 미치는 Ganciclovir 및 Vidarabine의 병용효과에 관한 연구 (Combined Effect of Ganciclovir and Vidarabine on the Replication, DNA Synthesis, and Gene Expression of Acyclovir-resistant Herpes Simplex Virus)

  • 양영태;정동균;모리 마사가즈
    • 대한약리학회지
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    • 제25권1호
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    • pp.115-134
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    • 1989
  • Ganciclovir(GCV)와 Vidarabine(ara-A)을 단독으로 또는 동시에 HSV-1에 작용시켰을때 HSV-1의 복제, DNA합성 및 단백질 합성에 미치는 영향을 관찰할 목적으로 본 연구를 시행하였다. 본 실험에서는 4가지의 다른 HSV-1(Wild type KOS, $VCV^r$, $IUdR^r$, 및 $PAA^r5$)를 사용하였다. Virus복제에 미치는 항 virus약의 효과는 Vero 세포단층 배양에서 Yield reduction assay에 의해서 관찰하였다. 항 virus약의 virus DNA 합성에 미치는 영향은 NaI 밀도 구배초원심침전후 $H^3-$표지 virus DNA의 방사능에 의해서 관찰하였다. Virus 단백질의 합성에 미치는 항 virus약의 효과는 $^{35}S$를 표지한 후 polyacrylamide 겔 전기영동법, 자가방사기록법 그러고 virus bands의 음영농도주사법을 이용, 관찰하여 다음과 같은 결과를 얻었다. 1. GCV는 wild trype HSV-1 KOS와 $PAA^r5$의 복제를 강력하게 억제하였으나 $ACV^r$$IUdR^r$는 GCV에 대해서 중등도의 저항을 보였다. ara-A는 실험대상의 모든 HSV-(KOS, $ACV^r$, $IUdR^r$$PAA^r5$)의 복제에 대해서 거의 비슷하게 억제효과를 보였다. GCV와 ara-A 동시첨가는 KOS와 $PAA^r5$의 복제에 대해서 상승적인 억제효과를 보였고 $ACV^r$$IUdR^r5$의 복제에 대해서는 상가작용이하의 억제 효과를 보였다. 2. GCV또는 ara-A는 HSV-1감염 Vero세포에서 virus DNA의 합성을 유의하게 억제하였다. GCV와 ara-A의 동시첨가는 KOS 또는 $PAA^r5$ 감염세포에서 virus DNA 합성을 GCV 또는 ara-A를 단독 첨가하였을때 보다 현저하게 억제하였다. $ACV^r$ 또는 $IUdR^r$ 감염세포에서는 이런 현상을 관찰할 수 없었다. 3. Wild type HSV-1 감염세포에서 virus-단백질의 합성은 GCV, ara-A의 단독 첨가 또는 GCV와 ara-A의 동시첨가에 의해서 변경되지 않았다. Wild type HSV-1 감염말기에 GCV 또는 ara-A 단독 혹은 동시첨가에 의해서 virus 단백질의 합성은 경미하나마 유의성있게 증가하였다. Wild type HSV-1과 $PAA^r5$에 의한 단백질의 합성은 GCV 또는 ara-A 단독 첨가에 의해서 유의성있게 억제되었다. GCV또는 ara-A의 동시첨가는 GCV또는 ara-A를 단독으로 첨가했을 경우보다 단백질의 합성을 더욱 억제하였다. 이상의 실험결과로 보아 GCV와 ara-A의 동시사용은 HSV-1 혹은 ACV저항 DNA polymerase변이주인 $PAA^r5$에 대해서 상승적인 억제작용을 나타냈으며 이 효과는 virus DNA 합성 억제에 의한 것으로 생각된다. ACV저항 thymidine kinase 변이주인 $ACV^r$$IUdR^r$에 대해서는 ara-A가 유효하였다. 항 virus 약물에 의한 virus 단백질합성의 변화는 virus DNA 합성에 대한 억제효과에 인한 것으로 사려된다.

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Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2015년도 춘계학술대회 및 임시총회
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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