• 한국미생물·생명공학회지
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• 제46권4호
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• pp.334-345
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• 2018

Spore-forming Bacillus species are commercially available probiotic formulations for application in humans. They have health benefits and help prevent disease in hosts by combating entero-pathogens and ameliorating antibiotic-associated diarrhea. However, the molecular and cellular mechanisms of these benefits remain unclear. Here, we report the draft genome of a potential probiotic strain of Bacillus clausii B106. We mapped and compared the probiotic profile of B106 with other reference genomes. The draft genome analysis of B106 revealed the presence of ADI pathway genes, indicating its ability to tolerate acidic pH and bile salts. Genes encoding fibronectin binding proteins, enolase, as well as a gene cluster involved in the biosynthesis of exopolysaccharides underscored the potential of B106 to adhere to the intestinal epithelium and colonize the human gut. Genes encoding bacteriocins were also detected, indicating the antimicrobial ability of this isolate. The presence of genes encoding vitamins, including Riboflavin, Folate, and Biotin, also indicated the health-promoting ability of B106. Resistance of B106 to multiple antibiotics was evident from the presence of genes encoding resistance to chloramphenicol, ${\beta}$-lactams, Vancomycin, Tetracycline, fluoroquinolones, and aminoglycosides. The findings indicate the significance of B. clausii B106 administration during antibiotic treatment and its potential value as a probiotic strain to replenish the health-promoting and disease-preventing gut flora following antibiotic treatment.

• Genomics & Informatics
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• 제19권4호
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• pp.43.1-43.12
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• 2021

Campylobacter jejuni is one of the most prevalent organisms associated with foodborne illness across the globe causing campylobacteriosis and gastritis. Many proteins of C. jejuni are still unidentified. The purpose of this study was to determine the structure and function of a non-annotated hypothetical protein (HP) from C. jejuni. A number of properties like physiochemical characteristics, 3D structure, and functional annotation of the HP (accession No. CAG2129885.1) were predicted using various bioinformatics tools followed by further validation and quality assessment. Moreover, the protein-protein interactions and active site were obtained from the STRING and CASTp server, respectively. The hypothesized protein possesses various characteristics including an acidic pH, thermal stability, water solubility, and cytoplasmic distribution. While alpha-helix and random coil structures are the most prominent structural components of this protein, most of it is formed of helices and coils. Along with expected quality, the 3D model has been found to be novel. This study has identified the potential role of the HP in 2-methylcitric acid cycle and propionate catabolism. Furthermore, protein-protein interactions revealed several significant functional partners. The in-silico characterization of this protein will assist to understand its molecular mechanism of action better. The methodology of this study would also serve as the basis for additional research into proteomic and genomic data for functional potential identification.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.127-127
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• 2017

Aluminum is the most abundant metallic element in the Earth's crust and considered as the most limiting factor for plant productivity in acidic soils. The inhibition of root growth is recognized as the primary effect of Al toxicity. Seeds of wheat cv. Keumkang (Korean cultivar) were germinated on petridish for 5 days and then transferred hydroponic apparatus which was treated with $0{\mu}M$ $AlCl_3$ (control), $100{\mu}M$ $AlCl_3$ and $150{\mu}M$ $AlCl_3$ for 5 days. The length of roots, shoots and fresh weight of wheat seedlings were decreased under aluminum stress. The concentrations of $K^+$, $Mg^{2+}$ and $Ac^{2+}$ were decreased whereas $Al^{3+}$ and $P_2O_5{^-}$ concentration was increased under aluminum stress. Using confocal microscopy, the fluorescence intensity of aluminum was increased with morin staining. In this study, a proteome analysis was performed to identify proteins, which is responsible to aluminum stress in wheat roots. In 10-day-old seedlings, proteins were extracted from roots and separated by 2-DE, stained by CBB. Using image analysis, a total of 47 differentially expressed protein spots were selected, whereas 19 protein spots were significantly up-regulated such as s-adenosylmethionine, oxalate oxidase, malate dehydrogenase, cysteine synthase, ascorbate peroxidase and 28 protein spots were significantly down-regulated such as heat shock protein 70, o-methytransferase 4, enolase, amylogenin by aluminum stress following protein spots analyzed by LTQ-FTICR mass spectrometry. The results provide the global picture of Al toxicity-induced alterations of protein profiles in wheat roots, and identify the Al toxicity-responsive proteins related to various biological processes that may provide some novel clues about plant Al tolerance.

• 한국작물학회지
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• 제61권4호
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• pp.227-234
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• 2016

본 연구에서는 쌀의 저장 단백질 조성 분석을 통해 자포니카와 통일형 품종의 식미 특성 차이를 구명하고자 하였다. 쌀의 저장단백질 분석 결과, 통일형 품종은 자포니카 품종에 비해 알부민과 글로부린이 각 1.2배, 1.3배, 글루텔린의 서브유닛인 산성 글루텔린 ${\alpha}-2$의 비율이 약 1.3배 낮은 반면, 글루텔린의 서브유닛인 산성 글루텔린 ${\alpha}-1$ 비율은 1.7배 높은 특성을 나타냈다. 또한, 저장단백질 조성과 식미특성의 상관성 분석을 실시한 결과, 식미 총평은 알부민(R=0.495, p<0.01), 글로부린(R=0.567, p<0.01)과 정의상관을 보였으며 산성 글루텔린 유닛인 ${\alpha}-1$은 총평과 뚜렷한 부의 상관(R=-0.612, p<0.01)이 인정되었다. 식미 총평 이외에도 밥의 외관, 맛, 찰기, 질감과 관련하여 알부민은 정의 상관, ${\alpha}-1$ 글루텔린은 부의 상관관계를 나타냈다. 생태형에 따른 저장단백질 비율이 식미 특성에 미치는 영향을 구명하기 위해 저장단백질 비율과 식미 특성과의 상관 분석을 실시한 결과, 산성 글루텔린의 서브유닛인 ${\alpha}-1$, 알부민, 글로부린이 통일형 품종의 식미 특성에 영향을 주는 것으로 보이며 향후, 다양한 품종을 이용한 추가 연구를 통해 정밀한 해석이 필요할 것으로 생각된다.

• Parasites, Hosts and Diseases
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• 제56권5호
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• pp.409-418
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• 2018

Acanthamoeba spp. are free-living protozoa that are opportunistic pathogens for humans. Cysteine proteases of Acanthamoeba have been partially characterized, but their biochemical and functional properties are not clearly understood yet. In this study, we isolated a gene encoding cysteine protease of A. castellanii (AcCP) and its biochemical and functional properties were analyzed. Sequence analysis of AcCP suggests that this enzyme is a typical cathepsin L family cysteine protease, which shares similar structural characteristics with other cathepsin L-like enzymes. The recombinant AcCP showed enzymatic activity in acidic conditions with an optimum at pH 4.0. The recombinant enzyme effectively hydrolyzed human proteins including hemoglobin, albumin, immunoglobuins A and G, and fibronectin at acidic pH. AcCP mainly localized in lysosomal compartment and its expression was observed in both trophozoites and cysts. AcCP was also identified in cultured medium of A. castellanii. Considering to lysosomal localization, secretion or release by trophozoites and continuous expression in trophozoites and cysts, the enzyme could be a multifunctional enzyme that plays important biological functions for nutrition, development and pathogenicity of A. castellanii. These results also imply that AcCP can be a promising target for development of chemotherapeutic drug for Acanthamoeba infections.

• 치위생과학회지
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• 제1권1호
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• pp.7-11
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• 2001

Streptococcus mutans는 인간에게 치아우식증을 유발하는 주요 세균으로 구강내에서 치태(plaque)가 형성되고, 탄수화물을 분해하면서 발생되는 유기산에 의해서 생존에 위협을 받게 된다. 하지만 약산성 조건에 노출되어도 산을 경험한 S. mutans는 강산성의 조건에서도 생존할 수 있는 산 저항 및 적응능을 획득하게 된다. 이러한 산 저항능은 Salmonell enterica serovar Typhimurium, E. coli 그리고 Shigella flexneri 등에서 확인 되었던 acid tolerance response(ATR)과 유사한 기작에 의해서 얻어진 것을 알 수 있었다. 이 현상은 산성조건을 경험할 때 유도되는 protein이 주요한 역할을 하는 것을 chloramphenicol 처리에 의해서 알 수 있었다. 동시에 acid shock에 의해서 유도되는 protein 이외에 여러 가지 요소들이 복합적으로 이런 생존 능력을 갖게 하였을 것이라는 가정을 할 수 있게 되었다. 이 연구에 의해서 S. mutans의 혐기적 조건에서의 산 적응기작을 확인하였으며, 산 저항성에 관여하는 유전자의 돌연변이체를 이용한 실험이 추가적으로 필요하다. S. mutans가 구강 내에 생존하는데 필수적인 산 저항기작이 결국 치아우식증 유발의 병독소로 중요한 역할을 하기 때문에 치아우식증의 근본적인 해결을 위한 연구의 일환으로 반드시 시행되어야 할 것으로 생각된다.

• 생명과학회지
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• 제25권7호
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• pp.833-838
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• 2015

다제약제 내성을 가지는 슈퍼 박테리아, 곰팡이, 바이러스, 기생충 감염은 주요한 건강위협인자들이다. 하지만, 건강위협 상황을 극복하기위해 현재 약제의 대안들중 항생펩타이드를 들 수 있다. 항생펩타이드는 자연계 다양한 종에서 생산된다. 항생펩타이드는 작은 단백질로 어류, 양서류, 파충류, 식물 그리고 동물의 감염으로부터 다세포생명체를 보호하는 선천성 면역에 관여하고 있다. 1980년대 이후로 매년 항생펩타이드의 수가 증가하고 있다. 박테리아, 원생생물, 곰팡이, 식물, 동물로부터 동정된 2,000가지 이상의 항생펩타이드가 항생펩타이드 데이터베이스에 등록되어 있다. 이러한 항생펩타이드의 대부분은 11-50개의 아미노산으로 구성되어 있고 하전상태는 0에서 +7까지이며 소수성은 31-70%를 차지하고 있다. 본 보고는 항생펩타이드를 생물학적 원천, 생물학적 기능, 펩타이드 성질, 공유결합패턴, 3차구조등에 의해 분류하였다. 항생펩타이드의 기능은 항균작용외에 세포주화성과 같은 세포생물학적 활성에도 기능성을 가지고 있다. 더욱이 림프절 스트로마로부터 기원한 fibroblastic reticular cell (FRC)에 염증상황 유도시 항생펩타이드가 발현되는 것을 확인하였다. FRC로부터 유도된 항생펩타이드는 whey acidic protein (WAP) 도메인을 포함하고 있었다. 이것은 단백질 도메인에 의해서도 항생펩타이드를 분류 할 수 있다는 것을 제시한다.

• Journal of Ginseng Research
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• 제41권3호
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• pp.247-256
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• 2017

Background: KG-135, a standardized formulation enriched with Rk1, Rg3, and Rg5 ginsenosides, has been shown to inhibit various types of cancer cells; however, the underlying mechanisms are not fully understood. In this study, we explored its effects in A549 human lung cancer cells to investigate the induction of Forkhead Class box O3a (FOXO3a) and autophagy. Methods: Cell viability was determined by sulforhodamine B staining. Apoptosis and cell cycle distribution were analyzed using flow cytometry. The changes of protein levels were determined using Western blot analysis. Autophagy induction was monitored by the formation of acidic vesicular organelles stained with acridine orange. Results: KG-135 effectively arrested the cells in G1 phase with limited apoptosis. Accordingly, a decrease of cyclin-dependent kinase-4, cyclin-dependent kinase-6, cyclin D1, and phospho-retinoblastoma protein, and an increase of p27 and p18 proteins were observed. Intriguingly, KG-135 increased the tumor suppressor FOXO3a and induced the accumulation of autophagy hallmark LC3-II and acidic vesicular organelles without an increase of the upstream marker Beclin-1. Unconventionally, the autophagy adaptor protein p62 (sequestosome 1) was increased rather than decreased. Blockade of autophagy by hydroxychloroquine dramatically potentiated KG-135-induced FOXO3a and its downstream (FasL) ligand accompanied by the cleavage of caspase-8. Meanwhile, the decrease of Bcl-2 and survivin, as well as the cleavage of caspase-9, were also drastically enhanced, resulting in massive apoptosis. Conclusion: Besides arresting the cells in G1 phase, KG-135 increased FOXO3a and induced an unconventional autophagy in A549 cells. Both the KG-135-activated extrinsic FOXO3a/FasL/caspase-8 and intrinsic caspase-9 apoptotic pathways were potentiated by blockade of autophagy. Combination of KG-135 and autophagy inhibitor may be a novel strategy as an integrative treatment for cancers.

• BMB Reports
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• 제34권1호
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• pp.73-79
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• 2001

Transcriptional activation domains are known to be inherently "unstructured" with no tertiary structure. A recent NMR study, however, has shown that the transactivation domain in human p53 is populated with an amphipathic helix and two nascent turns. This suggests that the presence of such local secondary structures within the overall "unstructured" structural framework is a general feature of acidic transactivation domains. These pre-existing local structures in p53, formed selectively by positional conserved hydrophobic residues that are known to be critical for transcriptional activity, thus appear to constitute the specific structural motifs that regulate recognition of the p53 transactivation domain by target proteins. Here, we report the results of a NMR structural comparison between the native human p53 transactivation domain and an inactive mutant (22L,23W$\rightarrow$22R,23S). Results show that the mutant has an identical overall structural topology as the native protein, to the extent that the amphipathic helix formed by the residues 18T 26L within the native p53 transactivating domain is preserved in the double mutant. Therefore, the lack of transcriptional activity in the double mutant should be ascribed to the disruption of the essential hydrophobic contacts between the p53 transactivation domain and target proteins due to the (22L,23W$\rightarrow$22R,23S) mutation.

• 유기물자원화
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• 제5권2호
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• pp.47-56
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• 1997

피혁 제조시 발생되는 크롬을 함유한 피혁 고형폐기물인 shaving scraps의 단백질 자원화 가능성을 검토하기 위하여 알카리제로 MgO를 농도별로 처리하여 가수분해시켜 회수한 hydrolyzed protein의 특성은 다음과 같다. MgO 처리에 의한 shaving scrap의 알카리 가수분해방법에서 크롬이 불용화 되는 pH 8.0 이상을 유지하기 위해서는 MgO를 5% 이상 처리해야 하는 것으로 나타났으며, MgO 처리에 따른 alkaline condition하에서 shaving scrap의 용해도는 약 60% 수준으로서 낮았다. MgO 처리에 의해 shaving scraps으로 부터 회수한 hydrolyzed protein의 평균 분자량은 약 80~100 KD이었고, collagen protein을 구성하고 있는 glycine, alanine 및 proline이 대부분을 차지하고 있었으며, 산성 아미노산인 aspartic acid와 glutamic acid도 상당량 함유되어 있는 것으로 나타났다. 무기성분중 Mg, Ca 및 Na는 액체비료의 무기양분으로서는 다소 많은 양이 함유되어 있었으며, Cr은 약 30~40ppm으로서 원시료 (40,000~42,000ppm)에 비해 크게 감소되었다.