• Journal of Microbiology and Biotechnology
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• 제23권3호
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• pp.357-363
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• 2013

The identification of bacteriophage proteins on the surface of Lactobacillus rhamnosus Pen was performed by LC-MS/MS analysis. Among the identified proteins, we found a phage-derived major tail protein, two major head proteins, a portal protein, and a host specificity protein. Electron microscopy of a cell surface extract revealed the presence of phage particles in the analyzed samples. The partial sequence of genes encoding the major tail protein for all tested L. rhamnosus strains was determined with specific primers designed in this study. Next, RT-PCR analysis allowed detection of the expression of the major tail protein gene in L. rhamnosus strain Pen at all stages of bacterial growth. The transcription of genes encoding the major tail protein was also proved for other L. rhamnosus strains used in this study. The present work demonstrates the spontanous release of prophage-encoded particles by a commercial probiotic L. rhamnosus strain, which did not significantly affect the bacterial growth of the analyzed strain.

• Molecules and Cells
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• 제27권2호
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• pp.159-166
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• 2009

Myocardial ischemia-reperfusion injury is a medical problem occurring as damage to the myocardium following blood flow restoration after a critical period of coronary occlusion. Oxygen free radicals (OFR) are implicated in reperfusion injury after myocardial ischemia. The antioxidant enzyme, Cu, Zn-superoxide dismutase (Cu, Zn-SOD, also called SOD1) is one of the major means by which cells counteract the deleterious effects of OFR after ischemia. Recently, we reported that a PEP-1-SOD1 fusion protein was efficiently delivered into cultured cells and isolated rat hearts with ischemia-reperfusion injury. In the present study, we investigated the protective effects of the PEP-1-SOD1 fusion protein after ischemic insult. Immunofluorescecnce analysis revealed that the expressed and purified PEP-1-SOD1 fusion protein injected into rat tail veins was efficiently transduced into the myocardium with its native protein structure intact. When injected into Sprague-Dawley rat tail veins, the PEP-1-SOD1 fusion protein significantly attenuated myocardial ischemia-reperfusion damage; characterized by improving cardiac function of the left ventricle, decreasing infarct size, reducing the level of malondialdehyde (MDA), decreasing the release of creatine kinase (CK) and lactate dehydrogenase (LDH), and relieving cardiomyocyte apoptosis. These results suggest that the biologically active intact forms of PEP-1-SOD1 fusion protein will provide an efficient strategy for therapeutic delivery in various diseases related to SOD1 or to OFR.

• 한국식품영양과학회지
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• 제15권1호
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• pp.45-55
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• 1986

우리나라에서 건제품(乾製品)으로 많이 이용되고 있는 갈치와 가자미의 건조(乾燥) 및 저장시(貯藏時) 일어나는 지질(脂質) 및 산화지질(酸化脂質)과 단백질(蛋白質)과의 반응(反應)이 이들 어육(魚肉)의 in vitro protein digestibility저하(低下)에 미치는 영향과 이에 수반되는 available lysine의 불용화(不溶化) 및 효소(酵素) 불소화성(不消化性) 물질(物質) 생성(生成) 정도의 변화를 실험한 결과는 다음과 같다. 가자미 및 갈치 생시료(生試料)의 소화율(消化率)은 87.63, 86.08%이었고, 일건(日乾), 열건시료(熱乾試料)는 건조(乾燥) 중 $1{\sim}2%$정도 저하(低下)되었으나 탈지시료(脫脂試料)는 각각 89.15, 87.15%로 증가하였다. 저장(貯藏) 중의 소화율(消化率)은 모든 시료(試料)에서 저하(低下)되었고 특히 갈치 열건시료(熱乾試料)는 78.00%로 심한 감소를 나타내었다. TIS함량은 생시료(生試料)에서 가자미, 갈치가 각각 0.88, 0.96mg/g solid이었으며, 탈지시료(脫脂試料)는 건조(乾燥) 및 저장(貯藏) 중 TIS함량의 증가가 적었고 소화율(消化率)은 높게 나타났다. VBN 및 PoV는 저장 10일에 저장초기의 2배(倍)정도로 증가하였고, 저장 30일에는 급격히 감소되었다. 갈변물질(褐變物質)의 형성은 저장기간이 증가함에 따라 증가되었다. 가자미 및 갈치 지질(脂質)의 주요(主要) 구성지방산(構成脂肪酸)은 $C_{18:1},\;C_{16:0},\;C_{22:6}\;C_{16:1}$등이었으며 총불포화지방산(總不飽和脂肪酸)함량은 가자미 79.2%, 갈치 67.8%이었고 저장 중 포화지방산(飽和脂肪酸)이 증가하고 불포화지방산(不飽和脂肪酸)이 감소하는 경향을 보였다. Available lysine은 갈치가 가자미보다 많았으며 저장 30일에는 저장초기량의 8.23%가 각각 손실되었다. 이상에서 수산단백식품(水産蛋白食品)은 건조(乾燥) 저장(貯藏) 중 산화지질(酸化脂質)과의 반응(反應)으로 단백질(蛋白質)의 양적(量的) 손실(損失)을 초래하였고, 비효소(非酵素)적 갈변(褐變), 지질일단백질(脂質一蛋白質)의 불용성(不溶性) 복합체(複合體) 형성(形成), available lysine의 불용화(不溶化), TIS의 생성(生成), 어육(魚肉)내의 구성지방산(構成脂肪酸)과 아미노산의 변화(變化)로 in vitro protein digestibility 저하(低下)를 비롯하여 어육(魚肉)의 갈변(褐變), 풍미손상(風味損傷) 및 영양가손실(營養價損失)등의 어육(魚肉)의 품질저하(品質低下)를 초래하는 주요인(主要因)으로 분석되었다.

• Journal of Microbiology and Biotechnology
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• 제26권9호
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• pp.1629-1635
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• 2016

A novel bacteriophage, PBES 02, infecting Cronobacter sakazakii was isolated and characterized. It has a spherical head of 90 nm in diameter and a tail of 130 nm in length, and belongs to Myoviridae as observed under a transmission electron microscope. The major virion protein appears to be 38 kilodaltons (kDa) in size. The latent period of PBES 02 is 30 min and the burst size is 250. Infectivity of the phage remained intact after exposure to temperatures ranging from 4℃ to 55℃ for 1 h. It was also stable after exposure to pHs ranging from 6 to 10 for 1 h. The phage effectively removed contaminating Cronobacter sakazakii from broth infant formula. PBES 02 has a double-stranded DNA genome of 149,732 bases. Its GC ratio is 50.7%. Sequence analysis revealed that PBES 02 has 299 open reading frames (ORFs) and 14 tRNA genes. Thirty-nine ORFs were annotated, including 24 related to replication and regulation functions, 10 related to structural proteins, and 5 related to DNA packaging. The genome of PBES 02 is closely related to that of two other C. sakazakii phages, CR3 and CR8. Comparison of DNA sequences of genes encoding the major capsid protein revealed a wide geographical distribution of related phages over Asia, Europe, and America.

• Journal of Ginseng Research
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• 제44권4호
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• pp.603-610
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• 2020

Background: Depression is a common neuropsychiatric disease that shows astrocyte pathology. Ginsenoside Rf (G-Rf) is a saponin found in Panax ginseng which has been used to treat neuropsychiatric diseases. We aimed to investigate antidepressant properties of G-Rf when introduced into the L-alphaaminoadipic acid (L-AAA)-infused mice model which is representative of a major depressive disorder that features diminished astrocytes in the brain. Methods: L-AAA was infused into the prefrontal cortex (PFC) of mice to induce decrease of astrocytes. Mice were orally administered G-Rf (20 mg/kg) as well as vehicle only or imipramine (20 mg/kg) as controls. Depression-like behavior of mice was evaluated using forced swimming test (FST) and tail suspension test (TST). We observed recovery of astroglial impairment and increased proliferative cells in the PFC and its accompanied change in the hippocampus by Western blot and immunohistochemistry to assess the effect of G-Rf. Results: After injection of L-AAA into the PFC, mice showed increased immobility time in FST and TST and loss of astrocytes without significant neuronal change in the PFC. G-Rf-treated mice displayed significantly more decreased immobility time in FST and TST than did vehicle-treated mice, and their immobility time almost recovered to those of the sham mice and imipramine-treated mice. G-Rf upregulated glial fibrillary acidic protein (GFAP) expression and Ki-67 expression in the PFC reduced by L-AAA and also alleviated astroglial change in the hippocampus. Conclusion: G-Rf markedly reversed depression-like behavioral changes and exhibited protective effect against the astrocyte ablation in the PFC induced by L-AAA. These protective properties suggest that G-Rf might be a therapeutic agent for major depressive disorders.

• 생명과학회지
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• 제20권7호
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• pp.1005-1011
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• 2010

$\gamma$-aminobutyric acid (GABA)는 중추신경계에서 억제성으로 작용하는 주요한 신경전달물질이다. GABA 수송체(GAT)는 연접간격에 존재하는 GABA를 세포 내로 재 흡수하여 GABA의 농도를 조절한다. 그런데 GABA 수송체가 어떻게 조절되는지는 아직 밝혀지지 않았다. 본 연구에서는 효모 two-hybrid system을 사용하여 뇌의 주요 GABA 수송체인 GAT1의 C-말단과 특이적으로 결합하는 ubiquitin-specific protease 14 (Usp14)를 분리하였다. Usp14는 GABA 수송체 GAT1및 GAT2와는 결합하지만, 다른 GAT isoform과는 결합하지 않았다. GAT1과의 결합에는 Usp14의 C-말단부위가 필수적으로 관여함을 확인하였다. 또한 이 단백질간의 결합을 GST pull-down assay로 확인하였으며, 생쥐 뇌 균질액의 co-immunoprecipitation을 통하여 in vivo에서도 GAT1과 Usp14가 결합함을 확인하였다. 이러한 결과들은 Usp14가 GAT1과 결합하여 세포막에 존재하는 GAT1의 수를 조절하는 역할을 할 가능성을 시사한다.

• The Plant Pathology Journal
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• 제34권1호
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• pp.59-64
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• 2018

Bacteriophages of Acidovorax citrulli, the causal agent of bacterial fruit blotch, were isolated from 39 watermelon, pumpkin, and cucumber leaf samples collected from various regions of Korea and tested against 18 A. citrulli strains. Among the six phages isolated, ACP17 forms the largest plaque, and exhibits the morphology of phages in the Myoviridae family with a head diameter of $100{\pm}5nm$ and tail length of $150{\pm}5nm$. ACP17 has eclipse and latent periods of $25{\pm}5min$ and $50{\pm}5min$, respectively, and a burst size of 120. The genome of ACP17 is 156,281 base pairs with a G + C content of 58.7%, 263 open reading frames, and 4 transfer RNA genes. Blast search and phylogenetic analysis of the major capsid protein showed that ACP17 has limited homology to two Stentrophomonas phages, suggesting that ACP17 is a new type of Myoviridae isolated from A. citrulli.

• The Plant Pathology Journal
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• 제32권6호
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• pp.584-588
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• 2016

Several Bacillus species were isolated from rice field soils, and 16S rRNA gene sequence analysis showed that Bacillus cereus was the most abundant. A strain named BC1 showed antifungal activity against Rhizoctonia solani. Bacteriophages infecting strain BC1 were isolated from the same soil sample. The isolated phage PK16 had an icosahedral head of $100{\pm}5nm$ and tail of $200{\pm}5nm$, indicating that it belonged to the family Myoviridae. Analysis of the complete linear dsDNA genome revealed a 158,127-bp genome with G + C content of 39.9% comprising 235 open reading frames as well as 19 tRNA genes (including 1 pseudogene). Blastp analysis showed that the proteins encoded by the PK16 genome had the closest hits to proteins of seven different bacteriophages. A neighbor-joining phylogenetic tree based on the major capsid protein showed a robust clustering of phage PK16 with phage JBP901 and BCP8-2 isolated from Korean fermented food.