• Title/Summary/Keyword: crude extract

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Purification and Characterization of Hpa I endonuclease (Hpa I endonuclease의 정제와 특성)

  • Yoon, Ho Sup;Kang, Sun Chul;Yoo, Ouk Joon
    • Microbiology and Biotechnology Letters
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    • v.13 no.1
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    • pp.87-91
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    • 1985
  • Hpa I endonuclease from Haemophilus parainfluenzae has been purified of homogeneity and its physical and ezymatic properties have been studied. For the purification of the enzyme, Heparin agarose, SP-sephadex C-25, DEAE-sephadex A-50 and phosphocellulose chromatography columns were used. The denatured and reduced form of the enzyme is a monomer of molecular weight of $30,000{\pm}1,000$ as judged by 10% polyacrylamide gel electrophoresis containing 0.1% sodium dodesyl sulfate. Hpa I endonuclease was maximally active at neutral pH (7.0 to 7.5) in the presence of 50 mM NaCl.

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Antioxidant Activity of Flavonoids Isolated from Vitex rotundifolia (순비기나무(Vitex rotundifolia)로부터 분리한 플라보노이드 성분의 항산화 활성)

  • Kim, You-Ah;Lee, Jung-Im;Hong, Joo-Wan;Jung, Myoung-Eun;Seo, Young-Wan
    • Ocean and Polar Research
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    • v.33 no.3
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    • pp.255-263
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    • 2011
  • The aim of this investigation was to evaluate antioxidant activity of crude extracts from the halophyte Vitex rotundifolia, their solvent fractions, and isolated compounds (1-3). Antioxidant capacity was determined by measuring DPPH radical, and authentic $ONOO^-$ and $ONOO^-$ generated from 3- morpholinsydnonimine (SIN-1) in vitro as well as degree of occurrence of intracellular ROS, NO and GSH in mouse macrophage Raw 264.7 cells. From comparative analysis, MeOH extract, n-BuOH, and 85% aq. MeOH solvent fractions showed significant antioxidant effect in DPPH radical and $ONOO^-$ assay systems. Activity-guided purification of n-BuOH and 85% aq. MeOH fractions led to the isolation of flavonoids 1-3. Among them, compound 1 exhibited excellent antioxidant effect in all bioassay systems tested. On the other hand, compounds 2 and 3 revealed potent inhibitory effect against $ONOO^-$ generated from SIN-1, comparable with the positive control penicillamine.

Optimization of biodiesel production via methyl acetate reaction from cerbera odollam

  • Dhillon, Sandip Singh;Tan, Kok Tat
    • Advances in Energy Research
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    • v.4 no.4
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    • pp.325-337
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    • 2016
  • Cerbera Odollam (sea mango) is a proven promising feedstock for the production of biodiesel due to its high oil content. Fatty acid methyl esters (FAME) were produced as the final reaction product in the transesterification reflux condensation reaction of sea mango oil and methyl acetate (MA). Potassium methoxide was used as catalyst to study its reacting potential as a homogeneous base catalyst. The initial part of this project studied the optimum conditions to extract crude sea mango oil. It was found that the content of sea mango sea mango oil was 55%. This optimum amount was obtained by using 18 g of grinded sea mango seeds in 250 ml hexane. The extraction was carried out for 24 hours using solvent extraction method. Response surface methodology (RSM) was employed to determine the optimum conditions of the reaction. The three manipulated variables in this reaction were the reaction time, oil to solvent molar ratio, and catalyst wt%. The optimum condition for this reaction determined was 5 hours reaction time, 0.28 wt% of catalyst and 1:35 mol/mol of oil: solvent molar ratio. A series of test were conducted on the final FAME product of this study, namely the FTIR test, GC-FID, calorimeter bomb and viscometer test.

DIGESTIBILITY OF NEUTRALIZED UREA-TREATED RICE STRAW AND NITROGEN RETAINED IN CROSSBRED HOLSTEIN STEERS

  • Promma, S.;Tasaki, I.;Cheva-Isarakul, B.;Indratula, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.4
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    • pp.487-491
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    • 1994
  • The experiment was carried out to study the digestibility of nutrients in the neutralized urea-treated rice straw when it was fed singly or in combination with concentrates. A total of 8 crossbred Holstein steers were randomly allocated in a $4{\times}4$ Latin square design consisted of 4 treatments, in which the neutralized straw/concentrates ratio on DM basis varied as 100/0, 90/10, 80/20 and 70/30. The results indicated that the digestibility of DM, ether extract and NFE, and TDN and DE of the diets tended to increase with an increase in the level of concentrates. Regression analysis showed that the values of intercepts should be used for estimating DM digestibility, TDN and DE of neutralized straw, when it was fed with concentrates. Digestibilities of crude fiber, NDF and ADF tended to be higher when it was fed singly than when fed with concentrates. Digestibilities of organic matter and CP were not so much changed with the increasing level of concentrates. Although the animals singly fed the neutralized straw showed positive body weight gain and N-balance, it should be necessary to supplement the concentrates to get more body weight gain and N-balance in the crossbred Holstein steers.

Effects of Different Levels of Concentrate in Complete Rations on Nutrient Digestibilities and Ruminal Metabolites in Sheep and Growth Performance in Korean Native Bulls

  • Baik, M.G.;Ha, J.K.;Kim, W.Y.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.4
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    • pp.371-377
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    • 1997
  • Objective of the present study were to investigate effects of different levels of concentrate in complete rations on nutrient digestibilities and ruminal fermentation in sheep and growth performance in Korean native bulls. Increasing levels of concentrate (35, 50, 65, and 80% of complete rations) improved digestibilities of dry matter (DM), crude protein (CP) and ether extract (EE) without affecting digestibility of neutral detergent (NDF) and acid detergent fiber (ADF). Increasing levels of concentrate decreased ruminal fluid pH but increased concentrations of $NH_3-N$, propionic acid, and total volatile fatty acids (VFA). Both the disappearance rates of DM and nitrogen (N) in an in sacco study were linearly increased as the levels of concentrate in complete rations increased. Meanwhile, increasing levels of concentrate in complete rations improved growth rate and feed conversion ratio in Korean native bulls. In conclusion, the complete rations containing 80% concentrate showed better digestibility and energy supply than those of the lower levels (35, 50 and 65%) of concentrate of the rations, resulting in improved growth performance of Korean native bulls.

Studies on Higher Fungi in Korea (I) -Activity of Proteolytic Enzyme from Sarcodon aspratus (Berk) S. Ito- (한국산 고등균류에 관한 연구(제 1보) -능이버섯의 단백분해효소 활성-)

  • Eun, Jae-Soon;Yang, Jae-Heon;Cho, Duck-Yee;Lee, Tae-Kyu
    • Journal of Pharmaceutical Investigation
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    • v.18 no.3
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    • pp.125-131
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    • 1988
  • This study was undertaken to investigate the proteolytic enzyme from Neungee mushroom [Sarcodon aspratus (Berk) S. Ito]. The proteolytic activity of Neungee was higher than other several edible mushrooms under various pHs. The potency of proteolytic enzyme of Neungee was same as the digestive drugs containing protease. So the proteolytic activity of the enzyme was increased in neutral or weak alkaline pH, whose characteristics would be alkaline protease. The specific activity of the purified enzyme obtained by using Tris acryl CM-cellulose ion exchange increased 20 times as compared with that of the crude extract. The proteolytic enzyme was stable at room temperature, but decomposition was fast when incubated at higher temperature more than $40^{\circ}C$. The half life of the enzyme was longest in neutral pH and rate constant was increased in acidic or alkaline solution.

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Isolation, Identification and Determination of Antioxidant in Ginger (Zingiber officinale) Rhizome

  • Cho, Kang-Jin;Kim, Jin-Weon;Choi, In-Lok;Kim, Jung-Bong;Hwang, Young-Soo
    • Journal of Applied Biological Chemistry
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    • v.44 no.1
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    • pp.12-15
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    • 2001
  • The antioxidative compounds and antioxidant contents of ginger (Zingiber officinale) rhizomes were determined. Substances reextracted using ethyl acetate from crude methanol extract of fresh ginger rhizome were separated through thin layer chromatography. Ten phenolic antioxidative bands were visualized through color reactions using ferric chloride-potassium ferricyanide and 1,1-diphenyl-2-picrylbydrazyl (DPPH). The antioxidative compounds were purified through preparative TLC and high performance liquid chromatography (HPLC), among which, five antioxidants were identified as 4-, 6-, 8-. and 10-gingerols and 6-shogaol on the basis of their molecular weights determined through LC-MS. As shown in experiments using DPPH free radicals, 6-Gingerol and PT4-HP8 (unknown) were revealed to be more efficient than BHT (butylated hydroxy toluene). Contents of gingerols were determined through reverse phase HPLC. Total gingerol contents (sum of 6-,8-, and 10-gingerols) in rhizomes of different ginger varieties varied significantly. The HG55 (collected at Wanju district in Korea) and the HG52 (imported from Brazil) showed the highest gingerol contents.

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Purification and Application of Earthworm /alpha-Galactosidase by Affinity Chromatography (Galactomannan 이용에 관한 연구 : Affinity Chromatography에 의한 지렁이 유래 /alpha-Galactosidase의 정제 및 응용법)

  • 박귀근;정규훈;소림수행
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.298-303
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    • 1999
  • An $\alpha$-D-galactosidase ($\alpha$-D-galactoside galactohydrolase, EC 3. 2. 1. 22) from earthworm was purified by affinity chromatography using N-$\varepsilon$-aminocaproyl-$\alpha$-D-galactopyranosylamine coupled to sepharose and its properties were examined. The specific activity of the purified enzyme, tested with p-nitrophenyl-$\alpha$-D-galactopyranoside as substrate, was 314 units/mg protein, representing an 122-fold purification of the original crude extract. The final preparation obtained from by Sephadex G-25 chromatography showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight was determined to be 48,000 by SDS-polyacrylamide gel electrophoresis. The purified galactosidase was showed maximum activity at pH 4.5 and 4$0^{\circ}C$, and was stable in the pH and temperature ranges from 4.0 to 5.5 and 30 to 5$0^{\circ}C$, respectively. The enzyme activity was inhibited by Zn2+, Hg2+ and Co2+. When the purified $\alpha$-galactosidase treated to guar gum for 6 hour, gel-promoting property was increased. It was clear that enzymatic elimination of galactose from guar gum by purified $\alpha$-galactosidase would lead to a significant increase in gelation ability.

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Cloning and Heterologous Expression of the β-Galactosidase Gene from Bifidobacterium longum RD47 in B. bifidum BGN4

  • Park, Min Ju;Park, Myeong Soo;Ji, Geun Eog
    • Journal of Microbiology and Biotechnology
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    • v.29 no.11
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    • pp.1717-1728
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    • 2019
  • The gene encoding β-galactosidase was cloned from Bifidobacterium longum RD47 with combinations of several bifidobacterial promoters, and expressed in B. bifidum BGN4. Among the recombinant bifidobacteria, BGN4+G1 showed the highest β-galactosidase level, for which the hydrolytic activity was continuously 2.5 to 4.2 times higher than that of BGN4 and 4.3 to 9.6 times higher than that of RD47. The β-galactosidase activity of BGN4+G1 was exceedingly superior to that of any of the other 35 lactic acid bacteria. When commercial whole milk and BGN4+G1 were reacted, BGN4+G1 removed nearly 50% of the lactose in the milk by the 63-h time point, and a final 61% at 93 h. These figures are about twice the lactose removal rate of conventional fermented milk. As for the reaction of commercial whole milk and crude enzyme extract from BGN4+G1, the β-galactosidase of BGN4+G1 eliminated 51% of the lactose in milk in 2 h. As shown below, we also compared the strengths and characteristics of the strong bifidobacterial promoters reported by previous studies.

Extracellular Tannase from Aspergillus ochraceus: Influence of the Culture Conditions on Biofilm Formation, Enzyme Production, and Application

  • Aracri, Fernanda Mansano;Cavalcanti, Rayza Morganna Farias;Guimaraes, Luis Henrique Souza
    • Journal of Microbiology and Biotechnology
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    • v.29 no.11
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    • pp.1749-1759
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    • 2019
  • Aspergillus ochraceus biofilm, developed on an inert support, can produce tannase in Khanna medium containing 1.5% (w/v) tannic acid as the carbon source, at an initial pH of 5.0, for 72 h at 28℃. Addition of 0.1% (w/v) yeast extract increased enzyme production. The enzyme in the crude filtrate exhibited the highest activity at 30℃ and pH 6.0. At 50℃, the half-life (T50) was 60 min and it was 260 min at pH 6.0. In general, addition of detergents and surfactants did not affect tannase activity significantly. Tannase has potential applications in various biotechnological processes such as the production of propyl gallate and in the treatment of tannin-rich effluents. The content of tannins and total phenolic compounds in effluents from leather treatment was reduced by 56-83% and 47-64%, respectively, after 2 h of enzyme treatment. The content of tannins and total phenolic compounds in the sorghum flour treated for 120 h with tannase were reduced by 61% and 17%, respectively. Interestingly, the same A. ochraceus biofilm was able to produce tannase for three sequential fermentative process. In conclusion, fungal biofilm is an interesting alternative to produce high levels of tannase with biotechnological potential to be applied in different industrial sectors.