• Title/Summary/Keyword: Leu-7

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Studies on the processing of rapid fermented anchovy prepared with low salt contents by adapted microorganism. -2. Thermodynamic characteristics of microbial extracellular protease isolated from fermented fish paste- (미생물을 이용한 저식염 멸치젓의 속성발효에 관한 연구 -2. 젓갈에서 분리한 단백질분해효소의 열역학적 특성-)

  • Cha, Yong-Jun;Lee, Eung-Ho
    • Applied Biological Chemistry
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    • v.33 no.4
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    • pp.325-329
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    • 1990
  • This study was undertaken to determine thermodynamic characteristics of B. subtilis p-4 and B. licheniformis p-5 proteases isolated from fermented anchovy paste. $K_m$ values of two proteases for casein as a substrate were 0.38mM in p-4 protease and 0.18mM in p-5 protease, respectively. Denaturation constants($K_D$) of p-4 and p-5 proteases were $12.2{\times}10^{-5}/sec\;and\;19.0{\times}10^{-5}/sec\;at\;40^{\circ}C,\;and\;35.7{\times}10^{-5}/sec\;and\;46.3{\times}10^{-5}/sec\;at\;50^{\circ}C$, respectively. Activation energies($E_a$) of p-4 and p-S pmteases were 19.6 Kcal/mole and 15.2kcal/mole, respectively. Free energy of activation(${\Delta}G^*$), activation enthalpy(${\Delta}H^*$) and activation entropy(${\Delta}S^*$) at $40^{\circ}C$ were 23.21Kcal/mole, 18.98Kcal/mole and -13.50 eu, respectively for p-4 protease and 22.93Kcal/mo1e, 14.58Kcal/mole and -26.68 eu, respectively for p-5 protease. The major amino acids in p-4 protease(151 residues of amino acid) were Gly, Glu, Pro, Asp, Ser, Ala, Lys and Leu, while those in p-5 protease(247 residues of amino acid) were Gly, Glu, Asp, Ala and Leu. It may be concluded that heat denaturation of two proteases showed liner regression curve and p-5 protease was more sensitive to heat than p-4 protease.

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Enhancement of PHB depolymerase Activity from Alcaligenes faecalis T1 by DNA Shuffling (DNA shuffling을 이용한 Alcaligenes faecalis T1의 PHB depolymerase 활성 증진)

  • 신동성;이영하;남진식
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.76-82
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    • 2003
  • To prepare evolved PHB depolymerase with increased activity for PHB or P(3HB-co-3HV) compared to the activity of the original PHB depolymerase from Alcaligenes faecalis T1, random mutation of the cloned PHB depolymerase gene was performed by using a DNA shuffling method. A library of mutated PHB depolymerase genes from A. faecalis T1 was fused to the ice nucleation protein (INP) gene from Pseudomonas syringae in pJHCl 1 and approximately 7,000 transformants were isolated. Using M9 minimal medium containing PHB or P(3HB-co-3HV) as the carbon source, mutants showing alteration in PHB depolymerase activity were selected from the transformants. The PHB depolymease activity of the transformants was confirmed by the formation of halo around colony and the turbidity decrease tests using culture supermatants. The catalytic activity of PHB depolymerase of the best mutant II-4 for PHB or P(3HB-co-13 mol% 3HV) was approximately 1.8-fold and 3.2-fold, respectively, higher than that of the original PHB depolymerase. DNA sequence analysis revealed that three amino acid residues (Ala209Val, Leu258Phe, and Asp263Thr) were substituted in II-4. From the mutational analysis, it was presumed that the substitution of amino acids near catalytic triad to more hydrophobic amino acids enhance the catalytic activity of PHB depolymerase from A. faecalis T1.

The Relationship between Isoniazid Resistance and 463 CodonMutation of katG Gne in Mycobacterium Tuberculosis (결핵균 katG 유전자내 463 Codon 돌연변이와 Isoniazid내성 관계)

  • Park, Young-Kil;Shim, Myung-Sup;Cho, Sang-Hyun;Bai, Gill-Han;Kim, Sang-Jae
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.1
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    • pp.8-13
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    • 1996
  • Background: The 463 codon mutation of katG gene has been reported as an useful marker for the detection of isoniazid(INH) resistant strains of M. tuberculosis. This study aimed to elucidate relationship between 463 mutation in katG gene and INH resistance in M. tuberculosis. Method: DNA was extracted from 28 INH susceptible strains(MIC$\geq\;0.2{\mu}g/ml$ on the L$\ddot{o}$wenstein Jensen media) and used for amplification of 189bp fragment containing 463 codon by PCR. Amplified fragments were digested by restriction enzyme Msp I, analyzed by single strand conformation polymorphism(SSCP) in the MDE gel and sequenced to prove mutation. Result: Only 7(25%) out of 28 were digestible by restriction enzyme Msp I. The SSCP pattern of 21 strains were distinctly different from that of M. tuberculosis H37Rv. Msp I undigestible PCR fragment was substituted at 463 codon from Arg(CGG) to Leu(CTG). Conclusion: This finding clearly indicate no relationship between 463 codon mutation of the katG gene and INH resistance.

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Exploration of β-Glucuronidase Activity of Lactic Acid Bacteria Isolated from Kimchi (김치에서 분리된 젖산균의 β-glucuronidase 활성 탐색)

  • Kim, Eun-Jung;Shin, In-Ung;Kwun, Se-Young;Park, Eun-Hee;Yi, Jae-Hyoung;Kim, Myoung-Dong
    • Microbiology and Biotechnology Letters
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    • v.47 no.3
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    • pp.434-440
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    • 2019
  • Lactic acid bacteria (LAB) isolated from kimchi were studied for their ${\beta}$-glucuronidase activity. Among the 156 strains tested, 52 strains utilized glucuronic acid as a carbon source and their intracellular ${\beta}$-glucuronidase activities were significantly higher than their extracellular activities. Leuconostoc mesenteroides KFRI 73007 isolated from turnip kimchi exhibited the highest intracellular ${\beta}$-glucuronidase activity of $0.77{\pm}0.01U/mg$ protein, which was further increased to $1.14{\pm}0.01U/mg$ protein under optimized reaction conditions (pH 7, $37^{\circ}C$). The activity of ${\beta}$-glucuronidase was notably decreased by the addition of divalent cations, and glucuronic acid was the best carbon source to produce ${\beta}$-glucuronidase in Leu. mesenteroides KFRI 73007.

AN EXPERIMENTAL STUDY OF THE EFFECT OF THE METHYLCHLOROTETRACYCLINE ON GROWTH OF CORONAL SUTURE IN RATS (Demethylchlorotetracycline이 백서의 관상봉합 발육에 미치는 영향에 관한 실험적 연구)

  • Lee, June Kyu
    • The korean journal of orthodontics
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    • v.10 no.1
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    • pp.7-13
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    • 1980
  • To study some informations on the morphogenesis and developmental process of the coronal suture in rats, the author performed daily oral administrations of demethylchlorotetracycline, a kind of tetracycline group, in the amount of 30mg/kg of body weight to the female rats from the 7th day of pregnancy to the time of delivery. Microscopic evaluation was undertaken on the fetal rats in the experimental group. The subject of this experiment were defined to the fetal rats of each group at 1st, 3rd, 7th and 14th day after birth. All these fetal rats were sacrificed and the heads were removed. All the tissue sections were fixed with $10\%$ formalin, Bouin' and Carnoy' solution and then stained by Hematoxylin-Van Gieson stain, or Feulgen and Rossenbeck, Periodic acid Schiff, and prepared for alcian blue reaction. The results were as follows; 1. The directions of osteogenic fibers were arranged irregulary during first 3 days, but after the 7th day they tended to change radial directions like control group. 2. The density of deep stained cells by Feulgen-Rossenbeck reaction were shown leu in the experimental group than that in the control group in first 3 days, but there was shown no significant difference between both groups after the 7th day. 3. PAS reaction in early stage was generally negative in the experimental group unlike as in the control group, but diffuse reaction was observed in the loose middle zone like as in the control group after 14th day. 4. Alcian blue reaction was negative in cambial zone, and slightly positive in uniting zone compared with control group in early stage. After 14th day, however, there was observed a tendency of moderately positive reaction.

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Varietal Analysis and Quantification of Amino Acid in Mulberry Fruits (뽕나무 계통별 오디의 아미노산 함량 분석)

  • Kim Hyun-Bok;Kim Sun-Lim;Kang Seok-Woo
    • Journal of Sericultural and Entomological Science
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    • v.46 no.2
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    • pp.47-53
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    • 2004
  • To reveal the possibility as a source of functional food and to increase the usability of mulberry fruits, amino acid analysis with mulberry fruits was carried out with L-8800 Amino acid auto analyzer. We analyzed 16 amino acid components including 6 essential amino acids. According to the results, the mean content of total amino acid was 4,640.8 mg/100 g DW. 'Sabangso(II)' showed the highest total amino acid content (6,919.3 mg/100 g DW) among the fifty accessions tested, whereas 'Gukkwang' showed the lowest content (3,389.1 mg/100 g DW). The distribution rate of amino acid components decreased in the order of Glu ($20.1\%$) > Gly ($12.7\%$) > Asp ($12.5\%$) > Thr ($8.2\%$) > Ala ($7.9\%$) > Arg ($7.5\%$) > Pro ($6.1\%$) > Ser ($6.1\%$) > Val ($5.7\%$) > Leu ($5.3\%$) > Ile ($2.7\%$) > His ($2.5\%$) > Met ($0.9\%$) > Tyr ($0.8\%$) > Lys ($0.7\%$) > Cys ($0.3\%$). 'Sabangso(II)' showed the highest content in 11 amino acid components, which were Glu (1,293.2 mg/100 g), Ser (390.2 mg/100 g), Thr (564.4 mg/100 g), His (175.3 mg/100 g), Arg (590.4 mg/100 g), Lys (53.5 mg/100 g), Gly (589.4 mg/100 g), Ala (499.3 mg/100 g), Leu (481.9 mg/100 g), Ile (240.9 mg/100 g), and Val (407.6 mg/100 g). Tyr and Cys were the highest in 'Jangsosang' (70.8 mg/100 g, 19.3 mg/100 g, respectively), and Asp, Pro, and Met were the highest in 'Jasan(I)', 'Hwansipchosaeng' and 'Palcheongsipyung(I)' (1,098.8 mg/100 g, 364.4 mg/100 g, 68.8 mg/100 g, respectively).

A Study on Components of Pollen Load (花紛荷(pollen load)의 성분조성에 관한 연구)

  • Kim, Jung-Hun;Lee, Kyu-Han;Kim, Eul-Sang;Park, Sung-Bae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.5
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    • pp.566-572
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    • 1992
  • Proximate composition and the contents of free sugars, minerals, fatty acids, amino acids and in vitro digestibility of mixed pollen load were analyzed. The protein in pollen load per 100g was 25.2g, lipid 5.4g, non-fibrous carbohydrate 50.0g and fiber 10.4. Free sugars in pollen load, as dry basis g per 100g, was fructose 26.6g, glucose 13.1g, sucrose 0.2g, maltose 1.3g, melezitose 0.02g and F/G ratio was 2.0. The content of mineral in pollen load was in the order K > P > Mg > Ca > Fe > Na > Mn > Zn > Cu. Unsaturated fatty acid of pollen load was about 69.1% and polyunsalturated fatty acid being about 50.3%. The ratios of SFA : MUFA : PUFA and PUFA : SFA were 0.61 : 0.37 : 1.00 and 1.63 respectively. The total amino acid contained in pollen load 22,337.5mg/100g and total essential amino acid was 39.2% of the total amino acid. proline was the most abundant amino acid (2,824.7 mg/100g),followed by Glu, Leu, Asp, Val, Phe, Ile, Ala and Thr. E/T ratio of protein in pollen load was 2.03 ; protein score, 65.2(Lys) : A/E ratio(egg), 61.7(Lys) : amino acid score, 51.7% after 1hr and on non-fibrous carbohydrate was 80.6%. But that of protein was 67.3% after 1hr, 75.5% after 2hr.

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Studies on the Drug Development of Coicis Semen(I) -Aminoacid Contents in Coicis Semen- (의이인(薏苡仁)(Coicis Semen)의 의약품개발(醫藥品開發)에 관(關)한 연구(硏究)(I) -아미노산(酸)의 함량(含量)에 대하여-)

  • Yong, Jae-Ick
    • Journal of Pharmaceutical Investigation
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    • v.7 no.1_4
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    • pp.1-12
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    • 1977
  • Free amino acid in ethanol extracts and total amino acid hydrolysates of Coicis semen were analyzed by amino acid autoanalyzer. The sample A (unpolished Coicis Semen) and sample B(polished Coicis Semen) are used in this experiments. The results obtained from this study are as follows: 1) 17 kinds of free amino acid (Asp, Thr, Ser, Glu, Pro, Gly, Ala, Val, Cys, Met, Ileu, Lew, Try, Phe, Lys, His, Arg,) including 7 kinds of essential amino acid (Val, Lew, Ileu, Thr, Lys, Met, Phe,) as human nutrition were identified and quantified but tryptophan. 2) Total free amino acids of sample A is more than about 3 folds that of sample B. 3) The distribution of free amino acids contained in sample A, threonine is the richiest and then comes Ala, Glu, Asp, and Pro, in that order. In sample B, glutamic acid is the richiest and then comes Thr, Asp, Ala, and Gly, in that order. 4) 17 kinds of total amino acid (Asp, Thr, Ser, Glu, Pro, Gly, Ala, Val, Cys, Met, Ileu, Lew, Tyr, Pher, Lys, including 7 kinds of essential amino acid (Val, Leu, Ileu, Thr, Lys, Met, Phe,) in human nutrition except tryptophan were identified and quanified. 5) Total amino acid content of sample A is more than about 1.06 folds that of sample B. 6) Total amino acid content of sample A in acid hydrolysates is more than about 1.06 folds that of sample B in acid hydrolysates. 7) Unknown chromatogram of ethanol extracts and acid hydrolysates of Coicis Semen were identified as Ornitine.

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A Study on the Amino Acid Contents of Edible Mushrooms (식용(食用)버섯류(類)의 아미노산(酸)에 관(關)한 연구(硏究))

  • Pyo, Myoung-Yun;Ro, Ihl-Hyeob
    • Journal of Nutrition and Health
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    • v.8 no.1
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    • pp.47-59
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    • 1975
  • Free amino acids in extracts and total amino acids in hydrolysates of eleven species of edible mushrooms were analyzed by amino acid autoanalyzer (Technicon PNC-1 Type). All these 11 species of mushroom can be repesented for convenience sake as follows. S-1; Agaricus campestris Fr. S-2: Agaricus campestris S-3; Pholiota nameko(I. Ito) S. Ito et Imai S-4; Auricularia auricula-judae(Fr.) $Qu{\acute{e}}l$ S-5; Tremella fuciformis Berk. S-6; Tricholoma matsutake(S. Ito et Imai) Sing. S-7; Pleurotus ostreatus Fr. $Qu{\acute{e}}l$ S-8; Lentinus edodes Berk Sing. S-9; Ramaria botrytis (Pers.) Ricken S-10; Coprinus comatus(Fr.) S.F, Gray S-11; Gyrophora esculenta The results obtained from this study are as follows. 1) 17 kinds of amino acid, including 7 kinds of essential amino acid in human nutrition except tryptophan were identified and quantified. 2) Of all free amino acids contained in mushrooms, glutamic acid is the richest, and then comes Ala, Thr, Pro and Lys in that order. There were no found Cys'and His in S-9;His in S-1; Met and Arg in S-11; Cys and Met in S-5;Pro, Cys, Met, Lys and Arg in S-4. Of all total amino acids which are closely related with nutritional valuation, glutamic acid is the richest, and then comes Asp, Ala, Arg, Leu, Thr, Gly in that order. Especially S-1 and S-2 contain high quantity $o{\acute{i}}$ proline in both free and total amino acids. 3) Cotents of ammonia in extracts of mushrooms in decreasing order in S-1, S-10, S-8, S-2, S-7, S-6, and S-2, S-6, S-8, S-9, S-1 in hydrolysates of mushrooms. 4) Gross Contents of free amino acid in extracts is high in decreasing order in S-10, S-1, S-7, S-6, S-8, and total amino acid in hydrolysates is high in S-10, S-2, S-2, S-8, S-1, S-9, S-6. 5) Besides 17 kinds of amino acid, 5 kinds of unknown amino acid are found in extracts and hydrolysates.

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Diketopiperazines from Cordyceps militaris (동충하초의 Diketopiperazine 성분)

  • Kim, Seon Beom;Hwang, Bang Yeon;Lee, Mi Kyeong
    • Korean Journal of Pharmacognosy
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    • v.44 no.4
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    • pp.336-343
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    • 2013
  • In a continuation of investigation on Cordyceps militaris, thirteen compounds were isolated from the $CH_2Cl_2$ and n-BuOH-soluble fraction of C. militaris. They were identified as twelve diketopiperazines such as cyclo($\small{L}$-Gly-$\small{L}$-Pro) (1), cyclo($\small{L}$-Ala-$\small{L}$-Pro) (2), cyclo($\small{L}$-Ser-$\small{L}$-Pro) (3), cyclo($\small{L}$-Val-$\small{L}$-Pro) (4), cyclo($\small{L}$-Thr-$\small{L}$-Pro) (5), cyclo($\small{L}$-Pro-$\small{L}$-Pro) (6), cyclo($\small{L}$-Thr-$\small{L}$-Leu) (7), cyclo($\small{L}$-Tyr-$\small{L}$-Ala) (8), cyclo($\small{L}$-Phe-$\small{L}$-Ser) (9), cyclo($\small{L}$-Phe-$\small{L}$-Pro) (10), cyclo($\small{L}$-Tyr-$\small{L}$-Pro) (11) and brevianamide F (13), and an amino acid, tryptophan (12). Their structures were identified on the basis of chemical evidences and spectroscopic analysis including 1D-NMR ($^1H$, $^{13}C$), 2D-NMR (HSQC, HMBC) and MS spectral data. Among the isolated compounds, compounds 1, 2, 6-11 are first reported from C. militaris.