• Title/Summary/Keyword: C3P

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Purification and Characterization of Lactate Dehydrogenase Isozymes in Channa argus (가물치(Channa argus) 젖산탈수소효소 동위효소들의 정제 및 특성)

  • Park, Eun-Mi;Yum, Jung-Joo
    • Journal of Life Science
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    • v.20 no.2
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    • pp.260-268
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    • 2010
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Channa argus were purified and characterized by biochemical, immunochemical and kinetic methods. The activity of LDH in skeletal muscle was the highest at 380.4 units and those in heart, eye and brain tissues were 13.4, 3,5 and 5.4 units, respectively. Citrate synthase (EC 4.1.3.7, CS) activity in heart tissue was the highest at 20.7 units. LDH/CS in skeletal muscle, heart, eye and brain tissues were 172.9, 0.6, 0.32 and 0.47. Protein concentration in skeletal muscle tissue was 14.7 mg/g and specific activities of LDH in skeletal muscle, heart, eye and brain tissues were 25.88, 0.79, 0.31 and 1.38 units/mg, respectively. Therefore, skeletal muscle tissue was anaerobic and heart tissue was aerobic. The LDH isozymes in tissues were identified by polyacrylamide gel electrophoresis, immunoprecipitation and Western blot with antiserum against $A_4$, $B_4$, and eye-specific $C_4$. LDH $A_4$, $A_3B$, $A_2B_2$. $AB_3$ and $B_4$ isozymes were detected in every tissue, $C_4$, $AC_3$, $A_2C_2$ and $A_3C$ were detected in eye tissue, and $A_3C$ was found in brain tissue. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ isozymes were purified by affinity chromatography and Preparative PAGE Cells. The LDH $A_4$ isozyme was purified in the fraction from elution with $NAD^+$ containing buffer of affinity chromatography. Eye-specific $C_4$ isozyme was eluted right after $A_4$, after which $B_4$ isozyme was eluted with plain buffer. As a result, one part of molecular structures in $A_4$, $B_4$ and eye-specific $C_4$ were similar, but were different from each other in $B_4$ and $C_4$. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The activity of LDH $A_4$, $A_2B_2$, $B_4$, and eye-specific $C_4$ isozymes remained at 39.98, 21.28, 19.67 and 16.87% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The $Km^{PYR}$ values were 0.17, 0.27 and 0.133 mM in $A_4$, $B_4$ and eye-specific $C_4$ isozymes, respectively. The optimum pH of LDH $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$, and $AB_3$ were pH 6.5, pH 8.5, pH 5.5, pH 6.0-6.5, pH 5.0 and pH 7.5. The $A_4$ and heterotetramer isozymes stabilized a broad range of pH. Especially, LDH activities in skeletal muscle tissue were high, resulting in a high degree of muscle activity.LDH metabolism in eye tissue seems to be converted faster from pyruvate to lactate by eye-specific $C_4$ isozyme as eye-specific $C_4$ have the highest affinity for pyruvate, and right after the conversion, oxidation of lactate was induced by $A_4$ isozyme. It was found that expression of Ldh-C, affinity to substrate and reaction time of $C_4$ isozyme were different according to the ecological environmental and feeding capturing patterns.

Identification and characteristics of Lactic Acid Bacteria Isolated from Nuruk (누룩으로부터 젖산세균의 분리 및 특성)

  • 이정훈
    • KSBB Journal
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    • v.15 no.4
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    • pp.359-365
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    • 2000
  • Three lactic acid bacteria (C-1 K-3 and T-1 strain) were isolated from Nuruk and characterized subsequently. They were useful strains for production of lactic acid and their growth was inhibited at 10% ethanol pH 4 These strains were identified as lactococcus lactis subsp. lactis NR C-1 Leuconostoc mesenteroides subsp. mesenterides NR K-3 and pediococcus pentosaceus NR T-1 respectively by morphological physiological and biochemical characterization Lac lactis subsp lactis NR C-1 showed the highest lactic acid productivity. Leu measenteroides subsp mesenteroides NR K-3 showed stable lactic acid productivity and its growth was inhibited at pH 4. P pentosaceus NR T-1 had lower lactic acid productivity than the other two bacteria but it could not grow at 10% ethanol pH 4 The lactic acid productivity of these three strains in MRS broth were higher than that in Skim milk media the optimum pH and temperature for the lactic acid production of the three strains were 30-32$^{\circ}C$ and pH 6.0∼6.8 Glucose was the optimal carbon souorce for the lactic acid production. In terms of antagonism lac lactis subsp lactis NR c-1 showed somewhat inhibitory efects against some Gram positive rod and cocci such as Lactobacillus brevis and Streptococcus mitis. And Leu mesenteroides subsp mesenteroides NR K-3 showed the inhibitory effects against Streptococcus mitis but P. pentosaceus NR T-1 didn't show any inhibitory effects against tested strains.

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Extraction and fractionation of proteins haying both chitinase and ${\beta}-1,3-glucanase$ canase activities from rice leaves ($Chitinase/{\beta}-1,3-glucanase$ 활성 동시보유 벼잎단백질 분획의 성질)

  • Uhm, Sung-Yon;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.36 no.5
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    • pp.370-375
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    • 1993
  • Five electrophoretic bands of crude enzyme extracted from rice leaves were found to possess both chitinase and ${\beta}-1,3-glucanase$ activities. These $chitinase/{\beta}-1,3-glucanase$ were resolved into acidic and basic fractions of protein by DEAE-cellulose and chitin affinity column chromatography. The optimal pH and temperature for ${\beta}-1,3-glucanase$ activity of two fractions were in the same extent as pH 5 and $60^{\circ}C$, whereas those for chitinase activity differed from one another; pH 3 and $60^{\circ}C$ for the acidic and pH 4 and $50^{\circ}C$ for the basic fraction, respectively. In addition, lysozyme activity was found in both fractions.

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Studies on Enzyme of the the Thermophilic Mold (PartV.) Purification of Xylanase (고온성 사상균의 효소에 관한 연구 (제5보) Xylanase의 정제)

  • Kim, Kwan;Kim, Yang-Hee;Jung, Dong-Hyo
    • Microbiology and Biotechnology Letters
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    • v.2 no.3
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    • pp.133-140
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    • 1974
  • 1) Two xylanase (designed as A and B) of Myriococcus albomyces were purified from an extract of wheat koji culture. Puriscation steps included first ammonium sulfate fractionation followed successively by SE-Sephadex column chromatgraphy. DEAE-Sephadex column chromatography and gel filtration on Sephadex G-100 repectively. 2) The optimum pH and pH stability for crude xylanse were found to be pH 5.0 and pH 4.0-7.0 respectively. 3) The optimium temperature was found to he 5$0^{\circ}C$ and for the thermal statbility of xylanase, the enryme incubated at $65^{\circ}C$ for 60min did not affect their stability. 4) The purised xylanase A and B were considered as liquefying xylanase and saccharogenic xylauase repectively. 5) The Bylanase A was most active at pH 4.0 and range of pH 3.0-8.0 at 3$0^{\circ}C$ for six hrs. The B was most active at pH 5.0 showing stability range of pH 4.0 to 8.5 at 3$0^{\circ}C$ for 6 hrs. incubation respectively. The Optimum temperature of xylanase A and B were found to be 7$0^{\circ}C$ and $65^{\circ}C$ for 60min repectively.

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AN EFFECT OF ND : YAG LASER ON THE BONDING STRENGTH OF COMPOSITE RESIN TO DENTIN AND PORCELAIN (Nd : YAG 레이저가 상아질 및 도재와 복합레진간의 결합강도에 미치는 영향)

  • Woo, Gum-Jin;Yang, Hong-So
    • The Journal of Korean Academy of Prosthodontics
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    • v.35 no.2
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    • pp.385-399
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    • 1997
  • The purpose of this experiment was to determine the effects of etching with a Nd : YAG Laser on dentin, or porcelain surface on the bond strength with composite resin. The dentin specimens were devided into the following 4 groups. D1 : No treatment D2 : Etched with 10% phosphoric acid D3 : Laser etchded with 1W, 20PPs D4 : Laser etched with 2W, 20PPS The procelain specimens were devided into the following 4 groups. P1 : diamond roughened P2 : etched with HF acid P3 : Laser etched with 2W, 20PPS P4 : Laser etched with 3W, 20PPS All specimens were veneered with resin. One half of the specimens were stored in $37^{\circ}C$ water for one day and the other half were thermocycled 1000 times at temperature of $5^{\circ}C\;to\;55^{\circ}C$ at 20 seconds intervals. After that, the bonding strength of composite resin to the dentin and porcelain was measured. The surface treated state and fractured state were observed with SEM. The following results were obtained. 1. In the dentin specimens, the bond strength of group D2 was highter than that of groups D1 and D3 in the case of the specimens stored in $37^{\circ}C$ water for one day, there was a statistically significant difference between group D2 and D1, D3 (P<0.05). The bonding strength of the specimens that were thermocycled decreased in the following order : group D2,D4,D3 and then D1. 2. In the porcelain specimens, the bonding strength of groups P1,P2 were higher than that of group P3 in the case of the specimens stored in $37^{\circ}C$ water for one day (P<0.05). The bonding strength of the specimens of being thermocycled decreased in the following order : group P2,P1,P4 and then P3. 3. The groups of high bond strength had a rougher surface and a high level of microporosity with SEM findings.

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Optimal Conditions for the Enzymatic Hydrolysis of Isolated Sesame Meal Protein (효소에 의한 참깨박 단백질의 최적 가수분해 조건)

  • Lee, S.H.;Cho, Y.J.;Kim, S.;Ahn, B.J.;Choi, C.
    • Applied Biological Chemistry
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    • v.38 no.3
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    • pp.248-253
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    • 1995
  • Optimum conditions for the enzymatic hydrolysis of isolated sesame meal protein were investigated. Optimum conditions by papain were $60^{\circ}C$, pH 6.0, 3% enzyme concentration to substrate and 1.5% substrate concentration, respectively. The optimum operating conditions using pepsin were $55^{\circ}C$, pH 9.0, 3% enzyme concentration to substrate and 1% substrate concentration. The optimum operating conditions using trypsin were $60^{\circ}C$, pH 9.0, 1% enzyme concentration to substrate and 1% substrate concentration.

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Detoxification and Paralytic Shellfish Poison Profile with Heating, Storage and Treatment of Alkaline in Blue Mussel, Mytilus edulis (알칼리 처리 및 가열, 저장에 따른 진주담치의 마비성 패류독 성분 특성 및 제독)

  • Jang, Jun-Ho;Yun, So-Mi;Lee, Jong-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.2
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    • pp.212-218
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    • 2006
  • Changes of paralytic shellfish poison (PSP) contents, toxicity and toxin composition with pH and storing periods at different temperature in toxic blue mussel, Mytilus edulis, were tested by using fluorometric HPLC method. Toxicity at pH 3 was the highest as 14.1 MU/g $(100\%)$ and showed 12.9 MU/g $(92.1\%)$ at pH 5, 9.0 MU/g $(63.8\%)$ at pH 7, 3.6 MU/g $(25.5\%)$ at pH 9 and 0.8 MU/g $(5.7\%)$ at pH 10 which suggested PSP was unstable at alkaline conditions. The decrease in toxicity during storage days was depend on pH and temperature. The toxicity markedly decreased until during the first S day storage $(19.9\~65.3\%)$ at all pH (3, 5, 7, 9) and temperature (30, 5, $-20^{\circ}C$), but, slightly decreased after then till to 30 days. C group toxin (C1 and C2) was the major components and other toxins such as GTX 1,2,3,4, STX and dcSTX were detected. Among the 8 toxins, GTX1,4, dcSTX and STX were firstly decreased according to the decreasing the toxicity at all processing conditions. The toxicity in blue mussel (14.1 MU/g) were able to remove by heating over 10 minutes at pH higher than 7.

Effects of Addition Level and Chemical Type of Propionate Precursors in Dicarboxylic Acid Pathway on Fermentation Characteristics and Methane Production by Rumen Microbes In vitro

  • Li, X.Z.;Yan, C.G.;Choi, S.H.;Long, R.J.;Jin, G.L.;Song, Man K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.1
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    • pp.82-89
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    • 2009
  • Two in vitro experiments were conducted to examine the effects of propionate precursors in the dicarboxylic acid pathway on ruminal fermentatation characteristics, $CH_4$ production and degradation of feed by rumen microbes. Fumarate or malate as sodium salts (Exp. 1) or acid type (Exp. 2) were added to the culture solution (150 ml, 50% strained rumen fluid and 50% artificial saliva) to achieve final concentrations of 0, 8, 16 and 24 mM, and incubated anaerobically for 0, 1, 3, 6, 9 and 12 h at $39^{\circ}C$. For both experiments, two grams of feed consisting of 70% concentrate and 30% ground alfalfa (DM basis) were prepared in a nylon bag, and were placed in a bottle containing the culture solution. Addition of fumarate or malate in both sodium salt and acid form increased (p<0.0001) pH of culture solution at 3, 6, 9 and 12 h incubations. The pH (p<0.0001) and total volatile fatty acids (VFA, p<0.05) were enhanced by these precursors as sodium salt at 3, 6 and 9 h incubations, and pH (p<0.001) and total VFA (p<0.01) from fumarate or malate in acid form were enhanced at a late stage of fermentation (9 h and 12 h) as the addition level increased. pH was higher (p<0.001) for fumarate than for malate as sodium salt at 3 h and 6 h incubations. Propionate ($C_3$) proportion was increased (p<0.0001) but those of $C_2$ (p<0.05) and $C_4$ (p<0.01 - p<0.001) were reduced by the addition of sodium salt precursors from 3 h to 12 incubation times while both precursors in acid form enhanced (p<0.011 - p<0.0001) proportion of $C_3$ from 6h but reduced (p<0.018 - p<0.0005) $C_4$ proportion at incubation times of 1, 3, 9 and 12 h. Proportion of $C_3$ was increased (p<0.05 - p<0.0001) at all incubation times by both precursors as sodium salt while that of $C_3$ was increased (p<0.001) from 6h but $C_4$ proportion was decreased by both precursors in acid form as the addition level increased. Proportion of $C_3$ was higher (p<0.01 - p<0.001) for fumarate than malate as sodium salt from 6 h incubation but was higher for malate than fumarate in acid form at 9 h (p<0.05) and 12 h (p<0.01) incubation times. Increased levels (16 and 24 mM) of fumarate or malate as sodium salt (p<0.017) and both precursors in acid form (p<0.028) increased the total gas production, but no differences were found between precursors in both chemical types. Propionate precursors in both chemical types clearly reduced (p<0.0001 - p<0.0002) $CH_4$ production, and the reduction (p<0.001 - p<0.0001) was dose dependent as the addition level of precursors increased. The $CH_4$ generated was smaller (p<0.01 - p<0.0001) for fumarate than for malate in both chemical types. Addition of fumarate or malate as sodium type reduced (p<0.004) dry matter degradation while both precursors in both chemical types slightly increased neutral detergent fiber degradability of feed in the nylon bag.

Fatty Acid Compositions of Some Seed Oils from the Pinaceae Family (솔과 식물 종실유의 지방산 조성)

  • Kim, Seong-Jin;Kim, Gee-Su;Yi, Min-Ok;Joh, Yong-Goe
    • Journal of the Korean Applied Science and Technology
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    • v.9 no.2
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    • pp.149-156
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    • 1992
  • Levels of total lipids in the seeds of three species of the Pinaceae family were determined and their fatty acid compositions were also analyzed by a gas-chromatograph equipped with a capillary column coated with Carbowax 20M. The results are summarized as follows: Lipid contents of the seeds amounted to 56.9% in P. koraiensis, 29.9% in P. thunbergii, and 21.2% in P. rigida. In all lipids 19${\sim}$20 fatty acid were detected and, surprisingly, fatty acids having ${\Delta}^5$-non-methylene interrupted conjugate double bond such as ${\Delta}^{5, 9}-C_{18:2},{\Delta}^{5, 9, 12}-C_{18:3}\;and\;{\Delta}^{5, 11, 14}-C_{20:3}$ occurred in appreciable amounts. In the lipids of P. koraiensis, the main component was $C_{18:2}{\omega}_6(45.0%)$, followed by $C_18:1{\omega} _9(26.9%)$ and ${\Delta}^{5, 9, 12}-C_{18:3}(14.6%)$, and then ${\Delta}^{5, 9}-C_{18:2}(2.2%)$ and ${\Delta}^{5, 11, 14}-C_{20:3}$ were also present. Levels of saturated fatty acid such as $C_{16:0}\;and\;C_{18:0}$ were as low as 7.5%. The seed oil of P. thunbergii predominantly comprised $C_{18:2}{\omega}_6(45.2%)$, and was then occupied by equal amounts ${\Delta}^{5, 9, 12}-C_{18:3}(18.1%)$ and $C_{18:1}{\omega}_9(18.1%)$. Its ${\Delta}^5, 11, 14}-C_{20:3}(5.8%)$ level was the highest in the samples tested. ${\Delta}^{5, 9}-C_{18:2}(2.8%)$ was also detected with other minor components. In the oils from the seeds of P. rigida, $C_{18:2}{\omega}_6$ was present as a main component, accompanied by $C_{18:1}{\omega}_9(21.6%)$ and ${\Delta}^{5, 9, 12}-C_{18:3}(20.3%)$. The latter showed higher level than in any other samples. A minor component corresponding to ${\Delta}^{5, 9, 12, 15}-C_{18:4}$(not confirmed by GC-Mass) occurred in P. thunbergii and P. rigida.