• 한국환경과학회:학술대회논문집
• /
• 한국환경과학회 2006년도 추계 학술발표회 발표논문집
• /
• pp.285-289
• /
• 2006

In screening of algicidal bacteria, we isolated a marine bacterium which had potent algicidal effects on harmful algal bloom (HAB) species. This organism was identified as a strain very close to Bacillus subtilisby 16S rRNA gene sequencing. This bacterium, Bacillus sp. SY-1, produces very active algicidal compounds against the harmful dinoflagellate Cochlodinium polykrikoides. We isolated three algicidal compounds (MS 1056, 1070, 1084) and identified them by amino acid analyses, fast atom bombardment mass spectrometry (FAB-MS), infrared spectroscopy (IR), $^1H$, $^{13}C$, and extensive two-dimensional nuclear magnetic resonance (2D NMR) techniques including $^1H-^{15}N$ HMBC analysis. One of them, MS 1056, contains a b-amino acid residue with an alkyl side chain of $C_{15}$. MS 1056, 1070, and 1084 showed algicidal activities against C. polykrikoides with an $LC_{50}$ (6 hrs) of 2.3, 0.8, $0.6\;{\mu}g/ml$, respectively. These compounds also showed significant algicidal activities against other harmful dinoflagellates and raphidophytes. In contrast, MS 1084 showed no significant growth inhibition against various organisms coexisting with HAB species in natural environments, including bacteria, eukaryotic microalgae, and cyanobacteria, although it inhibited growth of some fungi and yeasts. These observations imply that algicidal bacterium Bacillus sp. SY-1 and its algicidal compounds could play an important role in regulating the onset and development of HABs in the natural environments.

• 대한의생명과학회지
• /
• 제6권4호
• /
• pp.261-268
• /
• 2000

할미송이버섯으로부터 혈전용해효소 (FE-2)를 DEAE-cellulose, Mono-S column으로 분리 정제하였다. 이 효소는 분자량이 18.2 kDa 이었으며, ICP-MS (inductively coupled plasma mass spectrometer) 분석 결과 $Zn^{2+}$을 포함하고 있었다. 15번째까지의 N-terminal amino acid 서열은 A-L-Y-V-G-X-S-P-X-Q-Q-S-L-L-V이고, pH 7.5에서 활성이 가장 큰 염기성 단백질 분해효소로, EDTA와 1,10-phenanthroline에 의해 활성이 감소되는 metalloprotease였다. $Mg^{2+}$, $Zn^{2+}$, Fe$^{2+}$, Co$^{2+}$의 첨가 시 활성이 증가하였으나 Hg$^{2+}$의 경우 활성이 완전히 소멸되었다. 이 효소 (FE-2)는 단백질 분해효소 저해제인 E-64 (trans-epoxysuccinyl-L-leucylamido-(4-guanidino)-butane), PMSF (phenylmethylsulfonyl fluoride), pepstatin 과 2-mercaptoethanol의 영향을 받지 않으며, 섬유소원 (fibrinogen)과 반응 시 $A\alpha$ chain과 B$\beta$ chain은 분해시키지만 $\gamma$ chain과는 반응하지 않았다.

• 한국균학회지
• /
• 제28권1호
• /
• pp.60-65
• /
• 2000

할미송이버섯으로부터 3단계를 거처 분자량이 각각 18 kDa(FE-1)과 18.2 kDa(FE-2)인 두 개의 혈전용해효소를 분리 정제하였다. 이 효소들은 지금까지 알려진 혈전용해 효소의 질량에 비해 비교적 작았고 두 효소(FE-1과 FE-2)의 15번째까지의 N-terminal amino acid서열은 A-L-Y-V-G-X-S-P-X-Q-Q-S-L-L-V로 같았다. FE-1은 ICP-MS의 결과 $Zn^{2+}$을 mole당 0.98 mole 포함하고 있는 metalloprotease로 pH 7.5와 $55^{\circ}C$에서 가장 큰 활성을 나타내고, 1,10-phenanthroline의 첨가 시 저하된 활성은 $Zn^{2+}$나 $Co^{2+}$의 첨가에 의해 재생되었다.

• 대한임상검사과학회지
• /
• 제54권2호
• /
• pp.119-124
• /
• 2022

녹농균(Pseudomonas aeruginosa), 대장균(Escherichia coli) 및 포도알균(Staphylococcus aureus)은 기회감염균이다. 또한 이들 세균은 다제내성(Multiple-Drug Resistance, MDR) 세균의 성질을 가지는 것으로 알려져 있다. 녹농균, 대장균, 포도알균에 대한 다양한 효능을 지닌 6가지 발효액의 항균활성을 분석하였다. 발효액을 섭취했을 때 대장균에서 유전적 발현 변화가 일어나는지 알아보기 위해 annealing control primer를 사용하여 유전자발현 분석을 수행하였다. 디스크확산법을 통해 무화과식초와 대봉감식초가 다른 발효액에 비해 억제대의 크기가 가장 크게 나타났고, 토종약초발효소는 항균효과가 없었다. 대장균에 5% 무화과식초를 처리하여 21일간 매일 계대배양하여 Escherichia coli O157:H7 OmpW gene for outer membrane protein W 유전자 발현이 감소하며, Synthetic construct Lao1 gene for L-amino acid oxidase, complete cds 유전자가 발현이 증가함을 확인하였다. 발효액 중에서 특히 무화과식초를 섭취하는 것은 우리 주변에 항상 존재하는 대장균에 대해 방어능력을 더욱 단단하게 가질 수 있음을 의미하며, 나아가 다제내성균에 대한 천연치료제로 유용하게 쓰일 수 있기를 기대한다.

• 대한의생명과학회지
• /
• 제12권3호
• /
• pp.225-231
• /
• 2006

Fibrinolytic enzyme was purified from the fruiting bodies of Lepista nuda, using DEAE-Cellulose chromatography, Phenyl Sepharose chromatography, and Mono-S column chromatography. The substance has a molecular weight of 30006.62 Da as measured by MALD-TOF mass spectrometry. The N-terminal amino acid sequence of the enzyme was Tyr-Pro-Ser-Pro-Ser-His-Gln-Thr-Ala-Val-Asn-Ala-Ile-Ile-X. The activity of the enzyme was inhibited by PMSF, indicating that the enzyme is a serine protease. No inhibition was found with E-64, pepstatin, and EDTA. It has broad substrate specificity for synthetic peptides. The enzyme was stable up to $30^{\circ}C$. The enzyme hydrolyzes both Aa and y chains of human fibrinogen but did not show any reactivity for $B{\beta}$ chain of human fibrinogen.

• Archives of Pharmacal Research
• /
• 제16권3호
• /
• pp.231-236
• /
• 1993

Gly-224 residue of yeast alcohol dehydrogenase was mutated by site-directed mufagenesis to isoleucine, which is the corresponding amino acid residue of horse liver alcohol dehydrogenase. The mutated gene on M13 vector was subcloned in YEp13 and used to transform Saccharomyces cerevisiae 302-21 #2 strain, and the expressed protein was purified. The tumover numbers of mutant enzyme for ethanol and acetaldehyde were decreased copared to wild-type enzyme. The results of product inhibition studies indicated that the reaction mechanism was changed to Iso Theorell-Chance from Ordered Bi Bi. We supposed that Gly-224 was related to the enzyme reaction mechanism.

• Bulletin of the Korean Chemical Society
• /
• 제32권1호
• /
• pp.53-58
• /
• 2011

Cysteine and histidine-capped water-dispersible ZnS:Mn nanocrystals (ZnS:Mn-Cys and ZnS:Mn-His) were synthesized and their effects on E. coli and human cells were investigated. Particle sizes of these nanocrystals were found from HR-TEM images to be 3.5 nm and 4.0 nm, respectively. Their solution photoluminescence spectra showed identical broad emission peaks at 580 nm. ZnS:Mn-His significantly suppressed the growth of E. coli at $100{\mu}g/mL$ and 1 mg/mL concentrations, something not observed with ZnS:Mn-Cys. Consistent with this, greater inhibition of cell proliferation and viability were observed in HEK293 and IMR90 cells in ZnS:Mn-His at $100{\mu}g/mL$ and 1 mg/mL concentrations.

• Nutrition Research and Practice
• /
• 제9권6호
• /
• pp.592-598
• /
• 2015

BACKGROUND/OBJECTIVES: Increased mass of adipose tissue in obese persons is caused by excessive adipogenesis, which is elaborately controlled by an array of transcription factors. Inhibition of adipogenesis by diverse plant-derived substances has been explored. The aim of the current study was to examine the effects of the aqueous methanol extract of laver (Porphyra yezoensis) on adipogenesis and apoptosis in 3T3-L1 adipocytes and to investigate the mechanism underlying the effect of the laver extract. MATERIALS/METHODS: 3T3-L1 cells were treated with various concentrations of laver extract in differentiation medium. Lipid accumulation, expression of adipogenic proteins, including CCAAT enhancer-binding protein ${\alpha}$, peroxisome proliferator-activated receptor ${\gamma}$, fatty acid binding protein 4, and fatty acid synthase, cell viability, apoptosis, and the total content and the ratio of reduced to oxidized forms of glutathione (GSH/GSSG) were analyzed. RESULTS: Treatment with laver extract resulted in a significant decrease in lipid accumulation in 3T3-L1 adipocytes, which showed correlation with a reduction in expression of adipogenic proteins. Treatment with laver extract also resulted in a decrease in the viability of preadipocytes and an increase in the apoptosis of mature adipocytes. Treatment with laver extract led to exacerbated depletion of cellular glutathione and abolished the transient increase in GSH/GSSG ratio during adipogenesis in 3T3-L1 adipocytes. CONCLUSION: Results of our study demonstrated that treatment with the laver extract caused inhibition of adipogenesis, a decrease in proliferation of preadipocytes, and an increase in the apoptosis of mature adipocytes. It appears that these effects were caused by increasing oxidative stress, as demonstrated by the depletion and oxidation of the cellular glutathione pool in the extract-treated adipocytes. Our results suggest that a prooxidant role of laver extract is associated with its antiadipogenic and proapoptotic effects.

• Bulletin of the Korean Chemical Society
• /
• 제33권12호
• /
• pp.3943-3949
• /
• 2012

A series of aromatic hydrazides 3a-j were prepared by refluxing esters 2a-j with hydrazine hydrate in methanol, which were prepared by the esterification of 1a-j. Acetohydrazides 3a-j upon treatment with carbon disulfide and methanolic potassium hydroxide yielded potassium dithiocarbazate salts 4a-j, which on refluxing with hydrazine hydrate yielded substituted 4-amino-5-aryl-3H-1,2,4-triazole-3-thiones 5a-j. The target compounds 6a-j were synthesized by condensing furan-3-carboxylic acid in the presence of polyphosphoric acid under reflux. The structures of newly synthesized compounds were characterized by IR, $^1H$ NMR, $^{13}C$ NMR, elemental analysis and mass spectrometric studies. All the synthesized compounds were screened for their urease, acetylcholine esterase inhibition, antioxidant and alkaline phosphatase inhibition activity. Almost all of the compounds 6a-j showed good to excellent activities against urease and acetylcholine esterase more than the reference drugs. Compounds 6f and 6g were more potent scavenger of free radicals than the reference n-propyl gallate. Compound 6b and 6h showed excellent activities of alkaline phosphatase as compare to the reference $KH_2PO_4$.

• Biomolecules & Therapeutics
• /
• 제30권3호
• /
• pp.232-237
• /
• 2022

Autism spectrum disorder (ASD) having core characteristics of social interaction problems and repetitive behaviors and interests affects individuals at varying degrees and comorbidities, making it difficult to determine the precise etiology underlying the symptoms. Given its heterogeneity, ASD is difficult to treat and the development of therapeutics is slow due to the scarcity of animal models that are easy to produce and screen with. Based on the theory of excitation/inhibition imbalance in the brain with ASD which involves glutamatergic and/or GABAergic neurotransmission, a pharmacologic agent to modulate these receptors might be a good starting point for modeling. N-methyl-D-aspartic acid (NMDA) is an amino acid derivative acting as a specific agonist at the NMDA receptor and therefore imitates the action of the neurotransmitter glutamate on that receptor. In contrast to glutamate, NMDA selectively binds to and regulates the NMDA receptor, but not other glutamate receptors such as AMPA and kainite receptors. Given this role, we aimed to determine whether NMDA administration could result in autistic-like behavior in adolescent mice. Both male and female mice were treated with saline or NMDA (50 and 75 mg/kg) and were tested on various behavior experiments. Interestingly, acute NMDA-treated mice showed social deficits and repetitive behavior similar to ASD phenotypes. These results support the excitation/inhibition imbalance theory of ASD and that NMDA injection can be used as a pharmacologic model of ASD-like behaviors.