• 미생물학회지
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• 제54권2호
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• pp.152-154
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• 2018

Klebsiella pneumoniae는 그람 음성균에 속하고 막대 형태를 가지며 인간이나 동물의 폐에 감염하여 병을 일으키는 균이다. K. pneumoniae는 흔히 항생제 내성을 나타내는데 이로 인해 항생제를 통한 치료가 어려워지게 된다. 이런 상황에서 숙주 균에 특이적이고 민감하게 반응하는 박테리오파지는 항생제 내성균의 치료에 대한 대체적인 접근법으로 제안될 수 있다. 박테리오파지 KP1은 하수처리장에서 분리되었으며 K. pneumoniae에 대해 특정적인 감염성이 있다. 본 연구에서는 Klebsiella pneumoniae 박테리오파지 KP1의 유전체 초안 분석을 수행하였다. KP1의 유전체 초안은 167,989 bp의 길이, 39.6%의 G + C 비율로 구성되어있다. 295개의 예측된 ORF들과 14개의 tRNA 유전자를 가지고 있다. 또한 이들은 lysozyme, 그리고 holin과 같은 다양한 세포 용해 관련 효소들을 포함하고 있다.

• 미생물학회지
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• 제30권6호
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• pp.514-518
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• 1992

The present study has been perromed to investigate the optimal conditions for protoplast formation and regeneration of oleandomycin-producing Streptomyces antibioticus (S. antibioticus) KCTC 1081. Mycelia were grown in YME medium containing 0.2% (w/v) glycine and converted into the protoplast by incubating at 35.deg.C for 60 minutes in protoplast buffer (P buffer) containing 4 mg/ml lysozyme. The reversion of protoplasts to the normal filamentous state was examined by the growth on various synthetic agar media. A high reversion rate was obtained by incubating the protoplasts on a hypertonic agar medium containing 20 mM $Mg^{++}$, 5 mM $Ca^{++}$ and 0.3 M sucrose at 28.deg.C for 5 days. From these experiments, we established the improved regeneration medium and a protocol which supports higher and more consistent levels of regeneration of S. antibioticus protoplasts. The regenerant showed an increased antimicrobial activity compared with that of the initial strain.n.n.

• 미생물학회지
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• 제24권4호
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• pp.370-376
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• 1986

From the outer membrane portion of Gram-negative Klebsiella pneumoniae, the activity of 2-furaldehyde dehydrogenase depending upon beta-nicotinamide adenine dinucleotide was detected. Cytoplasmic membrane was preferentially extracted from crude membrane with $Mg^{2+}$ and Triton X-100, and then outer membrane was collected by ultracentrifugation. The crude enzyme was obtained by solubilization of outer membrane with lysozyme, ethylene diamine tetraacetate and Triton X-100. Thereafter 2-furaldehyde dehydrogenase was partially purified through column chromatography on QAE-Sephadex Q-50 and Sephadex G-150 and the enzyme activity was analyzed by means of high performance liquid chromatography. The optimal pH for the activity of the enzyme was about 9.5 and the optimal temperature was about $85^{\circ}C$. The partially purified enzyme retained tis activity at $85^{\circ}C$ for 5 hours. The optimal concentration of Triton X-100 for the activity of the enzyme was about 1.5% in the reaction mixture.

• BMB Reports
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• 제36권2호
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• pp.185-189
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• 2003

Chitinase (EC 3.2.1.14) was isolated from the culture filtrate of Streptomyces sp. M-20 and purified by ammonium sulfate precipitation, DEAE-cellulose ion-exchange chromatography, and Sephadex G-100 gel filtration. No exochitinase activity was found in the culture filtrate. The molecular mass of the purified chitinase was 20 kDa, estimated by a sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and was confirmed by activity staining with Calcofluor White M2R. Chitinase was optimally active at pH of 5.0 and at $30^{\circ}C$. The enzyme was stable from pH 4 to 8, and up to $40^{\circ}C$. Among the metals and inhibitors that were tested, the $Hg^+$, $Hg^{2+}$, and p-chloromercuribenzoic acid completely inhibited the enzyme activity. The chitinase activity was high on colloidal chitin, chitotriose, and chitooligosaccharide. The purified chitinase showed antifungal activity against Botrytis cinerea, and lysozyme activity against the cell wall of Botrytis cinerea.

• 한국수산과학회지
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• 제28권6호
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• pp.728-735
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• 1995

The peptidyl prolyl cis-trans isomerase(PPlase, EC 5.2.2.8) from Bacillus stearothermophilus SIC1 was extracted from the cells treated with by lysozyme. PPlase was purified from the cell extracts by heat treatment, ammonium sulfate precipitation, ion exchange chromatography and finally gel filtration (FPLC). The purity of purified the enzyme after Superose 12 column chromatography was examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The molecular weight of the purified PPlase was estimated as 18,000 by SDS-PAGE. The 39 amino acid residues from the N-terminus were determined by the protein sequencer. The enzyme showed the optimum pH at 8.0 and was stable at the range of pH 7.0 to 8.0. The enzyme was considerably stable after heat treatment at $60^{\circ}C$ for 30 minutes, and the enzyme was quite stable up to $65^{\circ}C$. The presence of the PPlase in the refolding solution accelerated the isomerization rate of the assay peptide.

• Journal of Microbiology
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• 제39권2호
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• pp.146-148
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• 2001

Lactobacillus crispatus ATCC 33820 and L. gasseri ATCC 33323 were grown in MRS broth (pH 6.5) at 37$^{\circ}C$ for 24 h and the antibacterial activities of cell free culture supernatants were determined by the agar well diffusion method. The culture supernatants were inhibitory to Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, Enterococcus faecalis, Pediococcus acidilacticii, and Lactobacillus helveticus. The supernatants did not show any lysozyme activity. Addition of catalase did not affect the antibacterial activities of the supernatants. The antibacterial substances were heat stable (100$^{\circ}C$ for 60 min) and sensitive to proteases.

• 한국식품위생안전성학회지
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• 제22권3호
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• pp.151-158
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• 2007

동치미로부터 분리한 유산균 (68 균주) 중 Escherichia coli O157에 대한 항균 효과를 나타내는 균주는 Lactobacillus plantarum K11로 동정되었다. 분리균주 La. piantarum K11이 생산한 박테리오신의 항균 활성은 대수증식기 후반부에 12,800 BU/mL로 최대 활성에 이르렀다. 항균 활성은 pepsin, protease, proteinase K, papain, chymotrypsin 및 trypsin 처리에 의해 완전히 소실되었으나, catalase, ${\alpha}-amylase$, lysozyme 및 lipase에 의해서는 영향을 받지 않았으므로 단백질성 물질임을 확인하였다. 게다가, 이 활성은 pH 3.0-9.0의 조건하에서나 -20, 4 및 $25^{\circ}C$에서 30일간의 저장 동안에도 안정하였다. 또한 $100^{\circ}C$에서 30분간 가열처리에도 비교적 안정한 편이었고, chloroform이나 hexane 처리에도 활성에 변함이 없었다. 분리 균주의 박테리오신은 Bacillus spp., Listeria spp. 및 Staphylococcus spp. 등의 일부 식중독균의 억제효과는 나타나지 않았으나, Enterobacter aerogenes와 E. coli 등의 장내세균의 억제에는 효과적이었으며, 특히 640 BU/mL의 박테리오신 처리에 의해서 10시간 배양만에 E. coli O157이 완전하게 사멸되었다.

• 한국미생물·생명공학회지
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• 제16권2호
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• pp.150-155
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• 1988

L-Lysine 생산균주인 Brevibacterium flavum ATCC 14067 과 Corynebacteriurn glutamicum ATCC13032에 섬유소 자화능이 우수한 Cellulomonas flavigena KFCC31221를 속간 융합시켜 섬유소로부터 L-lysine을 생산할 목적으로 이들 균주의 영양요구성 변이주 분리, 원형질체의 형성 및 재생의 조건을 조사하였다. NTG(500$\mu\textrm{g}$/$m\ell$)로 유기하여 penicillin-G(300$\mu\textrm{g}$/$m\ell$)로 농축한 변이주에서 B. flavum은 hse- str$^{r}$(100$\mu\textrm{g}$/$m\ell$), C. glutamicum는 Met$^{-}$T$hr^{-}$ Rif$^{r}$(5$\mu\textrm{g}$/$m\ell$), C. flavigena 는 T$hr^{-}$Val$^{-}$Kan$^{r}$(50$\mu\textrm{g}$/$m\ell$)의 gene marker를 가지는 영양요구성과 약제내성균주를 분리하였다. 공시균주의 원형질체 형성은 osmotic stabilizer로서는 0.5M sucrose, 배양기간은 대수증식기 중기의 균이 양호하였고, lysozyme 최적 처리 pH, 온도, 농도 및 반응시간은 각각 pH6.5, 33$^{\circ}C$, 500 $\mu\textrm{g}$/$m\ell$, 6시간으로 나타났으며, 이때의 원형질체형성율은 95-98%였다. 원형질체를 1.5% agar가 함유된 완전재생용 배지위에 0.7% agar가 함유된 완전재생용 배지로 중층했을 때 약 30~33%의 재생율을 보였다.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.767-771
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• 2005

Expression in yeast may prove more amenable to generating large amounts of viral antigens for a vaccine candidate. We, therefore, cloned the gene encoding the Hepatitis C virus (HCV) structural proteins (C-El-E2, c740) fused in-frame with, and immediately 3' to, the chicken-lysozyme signal peptide (C-SIG) gene and under the control of the yeast glyceraldehyde-3-phosphate dehydrogenase gene promoter. In yeast, the HCV structural proteins were expressed in two different forms: a processed and a nonprocessed aggregated form. Biophysical characterization by sucrose linear gradient centrifugation revealed that both forms were present in the same fractions with a buoyant density of 1.127-1.176 g/$cm^3$. These findings suggest that the efficient synthesis of HCV structural proteins in yeast may be an important tool to study virus assembly and may lead to the development of an HCV vaccine.

• Fisheries and Aquatic Sciences
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• 제23권1호
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• pp.2.1-2.6
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• 2020

Background: As the cost of fishmeal continues to rise, there will be a need to optimize the diet by minimizing dietary fishmeal inclusion in aquafeed. In this study, a 7-week experiment was conducted to evaluate soybean meal, fermented soybean meal (soytide), and sesame meal as fishmeal replacers in whiteleg shrimp, Litopenaeus vannamei. Methods: A 30%-based fishmeal diet was considered as control (CON), six other diets were prepared by replacing 20% or 40% of fishmeal with soybean meal (SB₂₀ and SB₄₀), fermented soybean meal (ST₂₀ and ST₄₀), or sesame meal (SM₂₀ and SM₄₀) from the CON diet. Twenty shrimp with average initial weight of 0.65 ± 0.05 g (mean ± SD) were randomly distributed into 21 tanks (45 L) and fed four times a day. Water temperature was controlled at 28 ± 1 ℃ and aeration was provided by air stones. Results: Weight gain, specific growth rate, feed efficiency, and protein efficiency ratio of shrimp fed CON showed no significant differences compared to shrimp fed all the other diets. However, growth performance of shrimp fed ST₂₀ diet was significantly higher than those of shrimp fed the SM₂₀ and SM₄₀ diets (P < 0.05). Superoxide dismutase activity (SOD) of shrimp fed CON, ST₂₀, and ST₄₀ diets was significantly higher than those of shrimp fed the SB₄₀ and SM₄₀ diets. But there were no significant differences among shrimp fed CON, SB₂₀, ST₂₀, ST₄₀, and SM₂₀ diets. Also, lysozyme activity of shrimp fed ST₂₀ diet was significantly higher than those of shrimp fed the SB₄₀ and SM₄₀ diets. Although, lysozyme activity of shrimp fed the CON diet was not significantly different compared to shrimp fed all the other experimental diets. Conclusions: Therefore, SB, ST, and SM could replace 40% of fishmeal based on growth performance and lysozyme. According to the SOD activity, SB and SM could replace 20% of fishmeal and ST could replace 40% of fishmeal in juvenile whiteleg shrimp Litopenaeus vannamei.