• Journal of Microbiology and Biotechnology
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• 제25권9호
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• pp.1493-1501
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• 2015

A Bacillus species, EMD4, with strong antibacterial activity was isolated from ganjang (soy sauce) and identified as B. subtilis. B. subtilis EMD4 strongly inhibited the growth of B. cereus ATCC14579 and B. thuringiensis ATCC33679. The antibacterial activity was stable at pH 3-9 but inactive at pH 10 and above. The activity was fully retained after 15 min at 80℃ but reduced by 50% after 15 min at 90℃. The activity was completely destroyed by proteinase K and protease treatment, indicating its proteinaceous nature. The bacteriocin (BacEMD4) was partially purified from culture supernatant by ammonium sulfate precipitation, and Q-Sepharose and Sephadex G-50 column chromatographies. The specific activity was increased from 769.2 AU/mg protein to 8,347.8 AU/mg protein and the final yield was 12.6%. The size of BacEMD4 was determined to be 3.5 kDa by Tricine SDS-PAGE. The N-terminal amino acid sequence was similar with that of Subtilosin A. Nucleotide sequencing of the cloned gene confirmed that BacEMD4 was Subtilosin A. BacEMD4 showed bactericidal activity against B. cereus ATCC14579.

• Journal of Microbiology and Biotechnology
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• 제12권6호
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• pp.972-978
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• 2002

Streptomyces sp. AD001 is a Gram-positive soil actinomycetes secreting an uncharacterized 2,4-dichlorophenol (DCP) oxidizing enzyme, whose activity is similar to the previously known Actinomycetes lignin-peroxidase (ALiP). This extracellular peroxidase was purified from Streptomyces sp. AD001 as a single protein band on an SDS-PACE by ammonium sulfate fractionation, Q-sepharose, concanavalin A, and Bio-Gel HTP column chromatographies. The molecular mass of the purified peroxidase was determined by SDS-PAGE to be 45.2 kDa, and 49.7 kDa with MALDI-TOF-MS, respectively. The highest level of peroxidase activity was observed at pH 7.5 and $30^{\circ}C$. The amino terminal sequence of the purified peroxidase (G-E-P-E-E-G-N-V-D-G-T-L) showed no significant homologies to my known proteins, suggesting that Streptomyces sp. AD001 may secrete a novel kind of bacterial peroxidase Initial rate kinetic data of the 2,4-DCP oxidation were best modeled with a random-binding bireactant system.

• 한국수산과학회지
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• 제21권2호
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• pp.85-96
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• 1988

Purification and some properties of alkaline proteinases in the pyloric caeca of skipjack, Katsuwonus vagans, were investigated. Four alkaline proteinases, temporarily designated proteinases I, II, III and IV, were identified from the tissue extract of the pyloric caeca by ammonium sulfate fractionation, DEAE-Sephadex A-50 chromatography, and Sephadex G-100 and G-200 gel filtration. Result of disc-polyacrylamide gel electrophoretic analysis showed that the purified proteinases II and III were homogenous with the yields of $1.5\%\;and\;1.2\%$, and those specific activities were increased to 33 to 37 fold over that of the crude enzyme solution, respectively. Molecular weight of the proteinases II and III determined by sephadex G-100 gel filtration were 28,500 and 24,200, respectively. The optimum conditions for the caseinolytic activity of the two enzymes were pH 9.6 and $48^{\circ}C$. The reaction rates of the two alkaline proteinases were constant to the reaction time to 80 min in the reaction mixture of $3.4{\mu}g/ml$ of enzyme concentration and $2\%$ casein solution. The Km values against casein substrate determined by the method of Lineweaver-Burk were $0.56\%$ for proteinase II and $0.30\%$ for proteinase II. The proteinases II and III were inactivated under the presence of $Ag^+,\;Hg^{2+},\;Ni{2+},\;Fe^{2+},\;and\;Cu^{2+}$, and but activated by $Mn^{2+}\;and\;Ca^{2+}$ and markedly inhibited by the soybean trypsin inhibitor and N-p-toluenesulfonyl-L-lysine chloromethyl ketone. Therefore, the proteinases II and III were found to be a group of serine proteases and assured to be trypsin-like proteinases.

• Journal of Microbiology and Biotechnology
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• 제21권5호
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• pp.483-493
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• 2011

An acid phosphatase (HppA) activated by $NH_4Cl$ was purified 192- and 34-fold from the periplasmic and membrane fractions of Helicobacter pylori, respectively. SDS-polyacrylamide gel electrophoresis revealed that HppA from the latter appears to be several kilodaltons larger in molecular mass than from the former by about 24 kDa. Under acidic conditions (pH${\leq}$4.5), the enzyme activity was entirely dependent on the presence of certain mono- and/or divalent metal cations (e.g., $K^+$,$ NH_4{^+}$, and/or $Ni^{2+}$). In particular, $Ni^{2+}$ appeared to lower the enzyme's $K_m$ for the substrates, without changing $V_{max}$. The purified enzyme showed differential specificity against nucleotide substrates with pH; for example, the enzyme hydrolyzed adenosine nucleotides more rapidly at pH 5.5 than at pH 6.0, and vice versa for CTP or TTP. Analyses of the enzyme's N-terminal sequence and of an $HppA^-$ H. pylori mutant revealed that the purified enzyme is identical to rHppA, a cloned H. pylori class C acid phosphatase, and shown to be the sole bacterial 5'-nucleotidase uniquely activated by $NH_4Cl$. In contrast to wild type, $HppA^-$ H. pylori cells grew more slowly. Strikingly, they imported $Mg^{2+}$ at a markedly lowered rate, but assimilated urea rapidly, with a subsequent increase in extracellular pH. Moreover, mutant cells were much more sensitive to extracellular potassium ions, as well as to metronidazole, omeprazole, or thiophenol, with considerably lowered MIC values, than wild-type cells. From these data, we suggest that the role of the acid phosphatase HppA in H. pylori may extend beyond 5'-nucleotidase function to include cation-flux as well as pH regulation on the cell envelope.

• Journal of Microbiology and Biotechnology
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• 제21권10호
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• pp.1021-1025
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• 2011

A Bacillus subtilis strain was isolated from the intestine of Sebastiscus marmoratus (scorpion fish) that was identified as Bacillus subtilis CH2 by morphological, biochemical, and genetic analyses. The chitosanase of Bacillus subtilis CH2 was best induced by fructose and not induced with chitosan, unlike other chitosanases. The strain was incubated in LB broth, and the chitosanase secreted into the medium was concentrated with ammonium sulfate precipitation and purified by gel permeation chromatography. The molecular mass of the purified chitosanase was detected as 29 kDa. The optimum pH and temperature of the purified chitosanase were 5.5 and $60^{\circ}C$, respectively. The purified chitosanase was continuously thermostable at $40^{\circ}C$. The specific acitivity of the purified chitosanase was 161 units/mg. The N-terminal amino acid sequence was analyzed for future study.

• Journal of Nutrition and Health
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• 제5권1호
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• pp.33-42
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• 1972

This study was designed to investigate the nutritional value of fermented soybean bran as animal feed. Natural soybean bran has low protein quality and high cellulase contents. The soybean bran was supplemented by urea and ammoniumsulfate as N-source for incubation of Aspergillus niger. After incubation of soybean bran with Aspergillus niger, the nutritional quality of protein and riboflavin contents were increased in general and more in aging process than in fermentation. In order to elucidate the biological efficiency of prepared soybean bran, 120 male weanling rats were divided into 22 groups, five rats each, and were fed by standard casein diet mixed with soybean bran in the proportion of 1/10, 1/15, 1/20 respectively. The animals were kept under the experimental diet for nine weeks. In the result of this study, food efficiency ratio showed higher in the groups of urea and ammonium-sulfate-add group than row soybean bran group but the former group is lower than the later in the body weight gains. Protein efficiency ratio was also same trend. It was noteworthy that the nitrogen retention rate in total body on the basis of urinary nitrogen excretion and dietary intake nitrogen and in big organ such as liver and spleen were higher in fermented group than raw soybean bran fed group. It was worth while to treat the soybean bran in first place fermentation and further aging process to elevate the biological efficiency and effect of nutritional values specifically of protein and of riboflavin.

• 폴리머
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• 제34권1호
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• pp.38-44
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• 2010

탄산칼슘/유기계 core-shell 입자를 제조하기 위해 core로는 침강성 탄산칼습을 사용하였고, shell로서는 여러 종류의 methyl methacrylate(MMA), ethyl acrylate(EA), n-butyl acrylate(BA), styrene(St), 2-ethylhexyl acylate(2-EHA)을 사용하여, 반응 온도 유화제, 개시제의 종류와 첨가량을 변화시킨 후에 유화중합을 시켜 최적의 유화중합 조건을 구하였다. 생성된 core-shell 입자의 구조는 FT-IR로, 입자크기와 형태는 입도 분석, SEM, TEM으로 각각 측정하였다. 그리고, 이 core-shell 입자에 부직포를 함침시켜 처리전후의 표면변화는 접촉각으로 확인하였다. 또한, 함침 처리 전후의 부직포/부직포 소재와 관능성 단량체 첨가한 부직포/부직포 소재의 박리 접착강도를 측정하여 상호 비교하였다. 탄산칼슘/유기 core-shell 입자 합성의 경우에는 유화제인 SDBS를 0.5 wt% 첨가한 탄산칼슘을 core로 하여 MMA와 0.1 wt% 농도의 APS를 단계적으로 주입하여 중합함으로써 탄산칼슘입자 표면에 단량체의 중합이 잘 유도될 수 있었으며 중합 도중에 새로운 중합체 입자의 생성이 적었다.

• 한국자원식물학회지
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• 제29권6호
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• pp.635-641
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• 2016

Sweet potato (Ipomoea batatas L.) shoot tips grown in vitro were successfully cryopreserved by encapsulation-vitrification. Encapsulated explants are very easily manipulated, due to the relatively large size of the alginate beads, and a large number of samples can be treated simultaneously. In this study, the effects of sucrose preculture, cryoprotectant preculture, and post-warm recovery media on regrowth, following liquid nitrogen (LN) exposure, were investigated to establish an efficient encapsulation-vitrification protocol for sweet potato. Shoot tips of plants grown in vitro were precultured in 0.3 M sucrose for 2 d before encapsulation. Encapsulated shoot tips were pre-incubated in liquid MS (Murashige and Skoog) medium containing 0.5 M sucrose for 16 h, before preculturing in sucrose-enriched medium (0.7 M sucrose) for 8 h. Shoot tips were osmoprotected with 35% plant vitrification solution 3 (PVS3) for 3 h, before being dehydrated with PVS3 for 2 h at $25^{\circ}C$. The encapsulated and dehydrated shoot tips were transferred to 2 mL cryotubes, suspended in 0.5 mL PVS3, and plunged directly into liquid N. High levels of shoot formation were obtained for the cv. Yeulmi (65.7%) and Yeonwhangmi (80.3%). The regrowth rates of cryopreserved samples in Yeulmi (78.9%) and Yeonwhangmi (91.3%), following culture on ammonium-free MS medium for 5 d, were much higher than those cultured on standard MS medium (65.7% and 80.3%, respectively). This encapsulation-vitrification is a promising method for the long-term preservation of sweet potato.

• Journal of Microbiology
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• 제35권2호
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• pp.109-116
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• 1997

An extracellular proteinase of Candida albicans was purified by a combination of 0~75% ammonium sulfate precipitation, DEAE Sepharose Fast Flow ion exchange chromatography, and Sephacryl S-200 HR molecular sieve chromatography. Its mlecular weight was approximately 41 kDa on SDS-PAGE and isoelectric point was 4.4. The enzyme was inhibited by pepstain A. Optimum enzyme activity ranged from pH 2.0 to 3.5 with its maximum at pH 2.5 and a temperature of 45$^{\circ}C$. The addition of divalent cations, $Ca^{2+}$, Zn$^{2+}$ and $Mg^{2+}$, resulted in no significant inhibition of enzymatic activity. However, some inhibitory effects were observed by Fe$^{2+}$, Ag$^{2+}$ and Cu$^{2+}$. With BSA as substrate, an apparent $K_m$ was determined to be 7$\times$10$^{-7}$ M and $K_i$, using pepstatin A as an inhibitor, was 8.05$\times$10$^{-8}$ M. N-terminal amino acid sequence was QAVPVTLXNEQ. Degradation of BSA and fibronectin was shown but not collagen, hemoglobin, immunoglobulin G, or lysozyme. The enzyme preferred peptides with Glu and Leu at the P$_1$ position, but the enzyme activity was highly reduced when the P$_2$ position was phe or pro. This enzyme showed antigenicity against sera of patients with candidiasis.

• 한국추진공학회:학술대회논문집
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• 한국추진공학회 2009년도 제33회 추계학술대회논문집
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• pp.177-180
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• 2009

본 연구에서는 HTPE 추진제 원료 및 HTPE 둔감 추진제 조성 2종에 대하여 DSC와 TGA를 사용하여 열분해 특성을 고찰하였다. AN이 포함된 HTPE 002는 약 $125^{\circ}C$에서 AN의 상전이 과정($II{\rightarrow}I$)을 거친 후, 약 $200^{\circ}C$범위까지 BuNENA와 AN이 함께 발열특성을 가지고 분해됨을 알 수 있었다. 추진제 HTPE 001과 HTPE 002의 열폭발에 대한 임계온도, $T_c$,를 Semenov의 열폭발 이론과 몇 가지 가열속도에서 측정된 비등온 곡선으로부터 계산되었고, 임계온도 계산에 사용된 열분해에 대한 활성화 에너지는 Kissinger방법으로 측정하였다.