• Title/Summary/Keyword: $Ca^{2+}$-EDTA

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Isolation and identification of Bacillus megaterium producing Alkaline ${\alpha}$-amylase

  • Jia, Shiru;Choe, Yong-Deok;Cho, Hoon
    • Journal of environmental and Sanitary engineering
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    • v.23 no.1
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    • pp.25-31
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    • 2008
  • A bacterial strain, Bacillus megaterium L-49 has been isolated and identified that produces alkaline ${\alpha}$-amylase. The cell is ellipsoidal, about $1.0-1.2{\times}3.0-3.6{\mu}m$ in diameter, Gram-positive, motile, and central partial central. Growth occurs in media containing 7% of NaCl. This strain could utilize D-glucose, lactose, xylose, sucrose, mannose, and maltose, and but it does not utilize D-fructose, and glycogen. Among the various concentrations of saturated ammonium sulfate, the retractation ratio in range of 70 to 100% was about 93%. However, in the case of acetone, it was about 98.7%. EDTA has activating effect and Ca2+ has no effect on alkaline ${\alpha}$-amylase activity. The alkaline ${\alpha}$-amylase has low thermal stability. The optimal temperature for reaction is $50^{\circ}C$. The alkaline ${\alpha}$-amylase activity maintained stabilizing at pH 6-11 and the optimal pH for reaction was 9-10.

Studies on the Immobilization of ${\beta}-Galactosidase$ from Bacillus subtilis (Bacillus subtilis ${\beta}-Galactosidase$의 고정화에 관한 연구)

  • Jang, Gi;Kim, Chang-Ryoul;Lee, Yong-Kyu
    • Korean Journal of Food Science and Technology
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    • v.22 no.4
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    • pp.426-433
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    • 1990
  • The conditions for immobilization of the partially purified ${\beta}-galactosidase$ form Bacillus subtilis HP4 and the properties of the immobilized enzyme have been investigated. The crude enzyme precipitated with cold acetone was purified about 68-fold through DEAE-cellulose and sephadex G-100 chromatography and its recovery was 19.9% The optimal conditions for Immobilization of enzyme were obtained in 2%(w/v) sodium alginate, 15%(v/v) enzyme solution and 2%(w/v) calcium chloride, and also the optimal stirring thme was 2 hours on the above conditions. The optimum temperature and pH values for immobilized enzyme were $55^{\circ}C$ and 6.5, respectively. Its residual activity was show 25% after heat treatment for an hour at $65^{\circ}C$, and found its high stability in pH 6.0 to 8.0. The enzyme activity was not affected b)· EDTA, 2-mercaptoethanol, KCN, protective agents, and other methal ions except Hg ion and Cu ion. The $K_m\;and\;V_{max}$ values of the immobilized enzyme on ONPG were $1.82{\times}10^{-2}M\;and\;3.57{\times}10^{-8}mole/min$, whereas those on lactose were $2.94{\times}10^{-2}M\;and\;1.68{\times}10^{-7} mole/min$, respectively. The remained enzyme activity for the immobilized enzyme was 95%t of original activity after storage of 40 days at $4^{\circ}C$, and when reused for 5 times was 81%. When skim milk(4.8% lactose) and 5% lactose solution were reacted with the immobilized enzyme(250 units/g) of lactose were 51% and 43%, respectively.

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Purification and Properties of Alkaline Protease from Streptomyce sp. YSA-130 (Streptomyces sp. YSA-130이 생산하는 Alkaline Protease의 정제 및 특성)

  • 윤성우;이강표;유주현;신철수;오두환
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.358-364
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    • 1989
  • A crystalline alkaline pretense- producing Streptomyce sp. YSA-130 was isolated from soil in alkaline medium(pH 10.5). The optimum culture condition of Streptomyce sp. YSA-130 for the production of alkaline protease was as follows; 2.0% soluble starch, 1.0% soytone, 0.3% $K_2$HPO$_4$, 0.02% MgSO$_4$.7$H_2O$, 0.8% Na$_2$CO$_3$, pH 10.5, 3$0^{\circ}C$, and 12 hr. The alkaline pretense from the culture broth of Streptomyce sp. YSA-130 was purified about 24 folds by ammonium sulfate precipitation , dialysis, DEAE-cellulose ion exchange chromatography, gel filtration on Sephadex G-15 and crystallization. Optimum temperature and pH of purified enzyme were 6$0^{\circ}C$, and 11.5. Temperature and pH stability of purified enzyme were 5$0^{\circ}C$, and 5.5-12.0. Calcium ion was effective to stabilize the enzyme at higher temperature. The molecular weight of the purified enzyme was approximately 30,000. The purified enzyme was inactivated by diisopropyl flurophosphate(DFP) but not affected by metal ion, EDTA, sulfhydryl reagent and stable detergent.

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Characterization of Nitroreductase Purified from TNT-degrading Bacterium, Pseudomonas sp. HK-6. (폭약 TNT를 분해하는 세균인 Pseudomonas SP. HK-6에서 분리정제된 Nitroreductase의 특성연구)

  • 호은미;강형일;오계헌
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.230-237
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    • 2004
  • In this study nitroreductase from Pseudomonas sp. HK-6 capable of degrading 2,4,6-trinitrotoluene (TNT) was characterized. Through a series of purification process including ammonium sulfate precipitation, DEAE-sepharose, and Q-sepharose, three different fractions I, II, and III having the enzyme activity of NTRs whose molecular weights were approximately 27 kDa were detected in fractions from HK-6 cells. Specific activity of the three fractions were approximately 4.85 unit/mg, 5.47 unit/mg, and 5.01 unit/mg, and concentrated to 9.0-, 10.1-, and 9.3-fold compared to crude extract, respectively. The optimal pH and temperature for the three NTR fractions were approximately 7.5 and $30^{\circ}C$, respectively. Metal ions, $Ag^{+}$ , $Cu^{ 2+}$, $Hg^{2+}$ inhibited approximately 70% of enzymes activities of all NTR, while $Fe^{2+}$ did not stimulate or inhibit the activities. Monitoring the effect of chemicals on the enzyme activity revealed that those NTR fractions lost enzyme activity in presence of $\beta$-mercaptoethanol, but were a little influenced by dithiothreitol, EDTA and NaCl. The three NTR fractions demonstrated enzyme activities for nitrobenzene and RDX as well as TNT.

최소가공 과일의 갈변저해를 위한 저해제의 탐색

  • 박연주;최소영;김영언;문광덕
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.04a
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    • pp.119.1-119
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    • 2003
  • 단체 급식의 증가와 더불어 편의식품을 선호하는 소비 경향에 맞추어 최소가공한 과일의 갈변저해를 위한 저해제를 탐색하였다. 실험에 사용한 과일은 배와 감으로 각각 박피하여 속심을 제거한 후 배는 8조각으로 절단하고 감은 1~2cm 두께로 slice 하였다. 저해제는 1% ascorbic acid, citric acid, NaCl, MgCl$_2$, CaCl$_2$, oxalic acid, EDTA, raffinose, salicylic acit, sporix를 사용하였다. 또한 열수 추출한 오약, 계피, 황기, 당기, 건칡, 진피, 천궁, 감초 등도 저해제로 사용하였다. 최소가공한 과일은 저해제에 3분간 침지한 후 물기를 제거하고 tray에 포장하여 실온에서 저장하면서 colorimeter를 이용하여 갈변도의 변화를 측정하였다. 갈변 정도를 나타내는 $\Delta$E 는 시간의 경과에 따라 점차 감소하는 경향을 보였으나 배의 경우 변화 정도가 크지 않았다. NaCl, MgCl$_2$, CaCl$_2$를 처리한 배의 갈변 정도가 가장 적었다. cysteine을 처리한 구에서는 황화합물 냄새가 났다. 또한 oxalic acid를 처리한 구에서는 연화가 빠르게 일어났으며 저장 시간이 경과함에 따라 수분이 용출되었다. 한약재는 자체 색으로 인해 배의 색변화를 증가시켰으며 이취가 발생했다. 또한 저장 시간이 경과함에 따라 건조가 빠르게 일어나 관능적으로도 좋지 않은 결과를 나타냈다. 감에서는 CaCl$_2$, MgCl$_2$를 처리한 구에서 색의 변화가 가장 적었으며 cysteine은 색변화는 적었으나 배의 경우과 같이 이취가 발생하였다. 한약제는 계피와 진피를 처리한 감에서 색의 변화를 저해하는 효과를 보였다.를 처리하였을 경우 당귀, 천문동을 처리한 구에서 효과를 보였다. oxalic acid는 갈변저해에는 효과적이었으나 시간이 경과함에 따라 수분의 용출이 증가하였으며 물성의 변화도 컸다. 또한 ZnCl$_2$를 처리한 구에서는 이취 발생과 수분 용출이 컸으며 색의 변화도 컸다.내었다. 고온 단시간 처리구의 경우 저온 장시간 처리구와는 달리 저장 1일 후부터 그 농도가 다소 상승되었다가 다시 감소는 경향을 보였다. 한편 열처리하여 냉각시킨 직후 및 저장1주 후 사과의 pH, 산도, 당도, 경도, 과육의 갈변도 등을 조사하였던 바 대조구 및 처리구간에 일부 항목에 있어서는 약간의 차이를 나타내었다. 대치할 수 있을 것이라고 사료된다. 수분함량은 기계적 검사보다 관능검사와 더욱 높은 상관관계를 나타냈다.내었다. 항균활성이 우수한 생약재를 농도별로 활성을 조사한 결과, 물 추출물과 10% Ethanol 추출물 모두 낮은 농도에서도 우수한 항균활성을 나타내었다.취와 함께 점질성 갈변물질이 생성되었다. 이와 같은 결과로 볼 때, BAAG의 처리는 BAAC의 경우보다 가격은 저렴하면서도 항균력은 우수한 천연 항균복합제재로써 농산물 식품원료에 적용하여 선도유지 기간을 연장할 수 있는 효과를 기대할 수 있었다. 과일 등의 포장제로서 이용할 가능성을 확인하였다.로 [-wh] 겹의문사는 복수 의미를 지닐 수 없 다. 그러면 단수 의미는 어떻게 생성되는가\ulcorner 본 논문에서는 표면적 형태에도 불구하고 [-wh]의미의 겹의문사는 병렬적 관계의 합성어가 아니라 내부구조를 지니지 않은 단순한 단어(minimal $X^{0}$ elements)로 가정한다. 즉, [+wh] 의미의 겹의문사는 동일한 구성요 소를 지닌 병렬적 합성어([$[W1]_{XO-}$ $[W1]_{XO}$ ]

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Antimicrobial, Antioxidant and Cellular Protective Effects of Houttuynia cordata Extract and Fraction (어성초 추출물 및 분획물의 항균, 항산화 및 세포보호활성)

  • Yun, Mid Eum;Lee, Ye Seul;Lee, Yun Ju;Park, Young Min;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.29 no.4
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    • pp.452-460
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    • 2018
  • This study was conducted to investigate the physiological activities of Houttuynia cordata extracts and fractions. H. cordata extracts were extracted with 50% ethanol and the ethyl acetate fractions were obtained from the extracts. Minimum inhibitory concentration (MIC) values of the ethyl acetate fraction for S. aureus and B. subtilis were $78{\mu}g/mL$ and $312{\mu}g/mL$, respectively, indicating the high activity against gram-positive bacteria. The free radical scavenging activity ($FSC_{50}$) for 1,1-diphenyl-2-picrylhydrazyl (DPPH) was higher in the ethyl acetate fraction with $12.00{\mu}g/mL$ compared to that of $27.15{\mu}g/mL$ for 50% ethanol extract. The total antioxidant activity ($OSC_{50}$) values for reactive oxygen species (ROS) produced in $Fe^{3+}-EDTA/H_2O_2$ system by a luminol-dependent chemiluminescence method were 2.91 and $0.983{\mu}g/ml$ for the 50% ethanol extract and ethyl acetate fraction, respectively. To investigate cellular protective effects on the HaCaT cell, the intracellular ROS scavenging activity was measured after UVB irradiation and the ethyl acetate fraction of H. cordata showed the activity in a concentration-dependent from $1.6{\mu}g/mL$ and a reduction rate of 54.3% at a maximum concentration of $12.5{\mu}g/mL$. Also, HaCaT cell protective effect against $H_2O_2$-mediated decreased the cell viability of the ethyl acetate fraction of H. cordata which significantly increased the cell viability from $0.8{\mu}g/mL$ and the maximum cell viability showed 86.9%. The ethyl acetate fraction of the H. cordata extracts was analyzed by TLC and HPLC. As a result, quercitrin, isoquercitrin, hyperoside, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, rutin and afzelin were identified. From the above results, it was suggested that the extracts and fractions of H. cordata have a potential to be applied in the field of cosmetics as a natural antioxidant/preservative capable of protecting the cell membrane from the oxidative stress by eliminating ROS and exhibiting the antimicrobial effect.

Antioxidant and Cellular Protective Effects of Moringa oleifera Leaves Extract (드럼스틱 잎 추출물의 항산화 및 세포보호 효과)

  • Xuan, Song Hua;Kim, A Rang;Jeong, Yoon Ju;Lee, Nan Hee;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.3
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    • pp.217-226
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    • 2016
  • In this study, we investigated the antioxidative and cellular protective effects on HaCaT cells and erythrocytes of Moringa oleifera (M. oleifera) leaves extract and its fractions. All experiments were performed with 50% ethanol extract, ethyl acetate fraction and aglycone fraction of M. oleifera leaves. The free radical scavenging activity ($FSC_{50}$) of the extract and fractions of M. oleifera leaves were in the following order: 50% ethanol extract ($77.10{\mu}g/mL$) < ethyl acetate fraction ($20.63{\mu}g/mL$) < aglycone fraction ($17.00{\mu}g/mL$) by using the 1, 1-diphenyl-2-picrylhydrazyl. In $Fe^{3+}-EDTA/H_2O_2$ system using the luminol, reactive oxygen species (ROS) scavenging activities (total antioxidant capacity, $OSC_{50}$) of aglycone fraction ($OSC_{50}=0.63{\mu}g/mL$) was the strongest among all extracts, which was much higher than L-ascorbic acid ($1.50{\mu}g/mL$). In the $^1O_2$-induced cellular damage of erythrocytes, the cellular protective effects of 50% ethanol extract (${\tau}_{50}=46.9min$) and aglycone fraction (${\tau}_{50}=122.1min$) were higher than (+)-${\alpha}$-tocopherol (${\tau}_{50}=37.7min$), known as a lipophilic antioxidant at $10{\mu}g/mL$. After cell damage induced by $400mJ/cm^2$ UVB irradiation, the cellular protective effects of ethyl acetate and aglycone fraction of M. oleifera leaves extract were showed on the concentration from 0.20 to $1.56{\mu}g/mL$. These results suggest that M. oleifera leaves extract and its fractions can function as a natural antioxidant agent in cosmetics on skin exposed to UV radiation by protecting cellular membrane against ROS.

Purification and Characterization of Chitinase from Antagonistic Bacteria Pseudomonas sp. 3098. (생물방제균 Pseudomonas sp. 3098이 생산하는 Chitinase의 정제 및 특성)

  • 이종태;김동환;도재호;김상달
    • Microbiology and Biotechnology Letters
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    • v.26 no.6
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    • pp.515-522
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    • 1998
  • Plant root rotting fungi, Fusarium solani are suppressed their growth by the chitinase which is produced from the antagonistic soil bacteria. The chitinase producable antagonistic bacterium Pseudomonas sp. 3098 was selected as a powerful biocontrol agent of F. solani from ginseng rhizosphere. The antagonistic Pseudomonas sp. 3098 was able to produce a large amount of extracellular chitinase which is key enzyme in the decomposition of fusarial hypal walls. The chitinase was purified from cultural filtrate of Pseudomonas sp. 3098 by the procedure of ammonium sulfate precipitation, anion exchange chromatography, gel filtration on Bio-Gel P-100, and 1st and 2nd hydroxyapatite chromatography. The molecular mass of the purified enzyme was ca. 45 kDa on SDS-FAGE. The optimal pH and temperature for the activity of purified chitinase were 5.0 and 45$^{\circ}C$, respectively. The enzyme was stable in pH range of 5.0 to 9.0 up to 5$0^{\circ}C$ The enzyme was significantly inhibited by metal compounds such as FeCl$_2$, AgNO$_3$ and HgCl$_2$, and was slightly inhibited by p-CMB, iodoacetic acid, urea, 2,4-DNP and EDTA. The enzyme had ability of digestion on colloidal chitin and chitin from shrimp shell, but could not digest chitosan and chitin from crab shell. Km value of the enzyme was 0.11% on colloidal chitin, and the maximum hydrolysis rate of the enzyme was 34% on colloidal chitin.

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Degradation and Detoxification of Disperse Dye Scarlet RR by Galactomyces geotrichum MTCC 1360

  • Jadhav, S.U.;Ghodake, G.S.;Telke, A.A.;Tamboli, D.P.;Govindwar, S.P.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.409-415
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    • 2009
  • Galactomyces geotrichum MTCC 1360 degraded the Scarlet RR(100 mg/l) dye within 18 h, under shaking conditions(150 rpm) in malt yeast medium. The optimum pH and the temperature for decolorization were pH 12 and $50^{\circ}C$, respectively. Enzymatic studies revealed an induction of the enzymes, including flavin reductase during the initial stage and lignin peroxidase after complete decolorization of the dye. Decolorization of the dye was induced by the addition of $CaCO_3$ to the medium. EDTA had an inhibitory effect on the dye decolorization along with the laccase activity. The metabolites formed after complete decolorization were analyzed by UV-VIS, HPLC, and FTIR. The GC/MS identification of 3 H quinazolin-4-one, 2-ethylamino-acetamide, 1-chloro-4-nitro-benzene, N-(4-chloro-phenyl)-hydroxylamine, and 4-chloro-pheny-lamine as the final metabolites corroborated with the degradation of Scarlet RR. The phytotoxicity study revealed the nontoxic nature of the final metabolites. A possible degradation pathway is suggested to understand the mechanism used by G. geotrichum and thereby aiding development of technologies for the application of this organism to the cleaning-up of aquatic and terrestrial environments.

Effect of Naturally Occurring Antioxidants on the Oxidative Stability of Fish Oil (천연 산화방지제가 어유의 산화안정성에 미치는 영향)

  • Han, Dae-Seok;Yi, Ock-Sook;Shin, Hyun-Kyung
    • Korean Journal of Food Science and Technology
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    • v.23 no.4
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    • pp.433-436
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    • 1991
  • Natural compounds which could improve the oxidative stability of fish oil was screened from spices, herbs and naturally occurring antioxidants. Induction period of fish oil determined from oxidation curve by Rancimat ($80^{\circ}C$) was hardly affected with the addition of water-soluble and lipid-soluble fractions of garlic, leek, sesame leave and orange peel, and of organic acids such as citric acid, EDTA and selenium. Caffeic acid, catechin, quercetin and gallic acid laurylester, however, could extend the induction periods by $2.2{\sim}3.8$ times with the addition level of 0.1%(w/w). Rosemary extract and sesamol have a marked effect in retarding oxidation of fish oil. For example, induction periods of the oil samples stabilized with 0.1 rosemary extract and 0.1 sesamol were 16.4 hr and 11.6 hr, respectively, as compared to 4.0 hr of a control. When rosemary extract was used in combination with ascorbic acid (0.02%) or 8-tocopherol (0.2%), induction period could be extended to ca. 28 hr due to the synergism.

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