• Title/Summary/Keyword: ${\alpha}$-amino acids

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The Biochemical Studies on Stored Soy-sauce (저장(貯藏)간장의 생화학적(生化學的) 연구(硏究))

  • Chang, Chi-Hyun
    • Applied Biological Chemistry
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    • v.9
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    • pp.9-27
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    • 1968
  • Studies were carried out in order to elucidate chemical components and microflora in three types of soy-sauce, 12-year aged soy-sauce prepared by improved method. 7-year aged and 20-year aged soy-sauce prepared by ordinary method. They results are summarized as follows: 1. The followings are found to be the important factors affecting the quality of soy sauce. a. Organic acids, reducing sugars and free amino acids were increased in the course of storage. b. In the aged soy-sauces under study non-volatile organic acid increased while volatile organic acid decreased and the total acidity was dependent only upon tie latter. c. It was found that suit concentration decreased during the storage. 2. The results of investigation of microflora in the stored soy-sauce are shown as follows. Soy-sauce Improved Ordinary Microbe 12-Y. 20-Y. 7-Y. Aerobic bacteria colony/1ml. 6 123 2 Halophilic lactic acid bacteria colony/1ml. 4 6 10 Osmophilic yeast colony/1ml. $828{\times}10^4$ 248 - b. In the stored soy-sauces, aerobic bacteria are incapable of growing due to drop in pH value and the influence of salt concentration. c. Halophilic lactic acid bacteria are incapable of growing due to drop in pH value, even the salt concentrations decreased during the storage. d. Osmophilic yeast are still growing in low pH value and in the decreasing salt concentration during the strage. 3. The results of amino acid analysis by paper partition chromatographic and calorimetric methods are shown as follows. a. Fourteen kinds of amino acid and thirteen amino acids were detected in the soy-sauce of 12-year aged improved soy-sauce and 7-year aged and 20-year aged ordinary one, respectively. b. The contents of aspartic acid, glutamic acid, serine, valine, leucine, lysine, histidine and methionine increased in the 20-year aged ordinary soy-sauce compared to the 7-year aged one. On the other hand those of alanine, tyrosine, phenylalanine and cystine decreased. 4. The results of sugar analysis by paper chromatography are as follows. a. In the 12-year aged improved soy-sauce, galactose, glucose, arabinose, xylose, rhamnose, maltose and an unknown were detected, and their amounts were in the above order except maltose and an unknown. b. Both in the 7 and 12-year aged ordinary soy-sauces, galactose, arabinose, xylose, glucose and rhamnose were detected and the amounts of the sugars were in the above order. c. In the non-aged ordinary soy-sauce, glucose was not detected but detected from 7-year and 20-year aged ordinary soy-sauce. 5. The results of organic acid analysis by paper chromatography were as follows. a. As volatile acids, acetic, propionic and butyric acids were detected in the 7-year aged ordinary soy-sauce. On the other hand in both the 20-year aged ordinary soy-sauce and the 12-year aged improved ones, only acetic acid was abundant while propionic and butyric acids were round in trace. It was found that propionic and butyric acids, as the unpleasant flavor components, decreased during the storage. b. In the ordinary soy-sauce, citric acid were produced during the storage and lack, malic and tartaric acids increased in the course of aging while succinic, glycolic, fumaric and malonic acids were shown to decrease. Glutaric and oxalic acids disappeared. Citric acid was produced also in the improved soy-sauce, but lactic, tartaric, succinic, malic, and glycolic acids decreased, while both malonic and glutaric acids disappeared. From the above results the citric acid production was considered to be a favorable factor for the taste. c. In the aged soy-sauces, pyruvic, α-ketoglutaric and probably acetoacetic and oxaloacetic acids (both in trace) were present and their amounts were in the above order. All of the α-keto acid abruptly decreased during the storage.

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Recombinant Expression and Characterization of Thermoanaerobacter tengcongensis Thermostable $\alpha$-Glucosidase with Regioselectivity for High-Yield Isomaltooligosaccharides Synthesis

  • Zhou, Cheng;Xue, Yanfen;Zhang, Yueling;Zeng, Yan;Ma, Yanhe
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1547-1556
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    • 2009
  • A novel thermostable $\alpha$-glucosidase (TtGluA) from Thermoanaerobacter tengcongensis MB4 was successfully expressed in E. coli and characterized. The TtgluA gene contained 2,253 bp, which encodes 750 amino acids. The native TtGluA was a trimer with monomer molecular mass of 89 kDa shown by SDS-PAGE. The purified recombinant enzyme showed hydrolytic activity on maltooligosaccharides, p-nitrophenyl-$\alpha$-D-glucopyranide, and dextrin with an exotype cleavage manner. TtGluA showed preference for short-chain maltooligosaccharides and the highest specific activity for maltose of 3.26 units/mg. Maximal activity was observed at $60^{\circ}C$ and pH 5.5. The half-life was 2 h at $60^{\circ}C$. The enzyme showed good tolerance to urea and SDS but was inhibited by Tris. When maltose with the concentration over 50 mM was used as substrate, TtGluA was also capable of catalyzing transglycosylation to produce $\alpha$-1,4-linked maltotriose and $\alpha$-1,6-linked isomaltooligosaccharides. More importantly, TtGluA showed exclusive regiospecificity with high yield to produce $\alpha$-1,6-linked isomaltooligosaccharides when the reaction time extended to more than 10 h.

Effects of Carbon Precursors and Hormones on the Lipogenesis and Lipolysis of Hanwoo Cattle Adipose Tissues

  • Lee, S.C.;Lee, H.J.;Kim, D.W.;Park, J.G.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.3
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    • pp.300-306
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    • 2000
  • This experiment was carried out to determine the contributions of acetate, glucose, amino acids and amino acid metabolites as carbon precursors for the incorporation of radioisotope, in intramuscular and subcutaneous adipose tissue and the effects of insulin on lipogenesis and adrenergic agent, norepinephrine on lipolysis in both tissues. The rate of incorporation of $C^{14}$ labelled acetate, glucose, leucine, isoleucine and ${\alpha}$-ketoisocaproic acid into adipose tissue has been measured in subcutaneous and intramuscular adipose tissues. The rate of incorporation was greater (p<0.05) from acetate than glucose in both subcutaneous and intramuscular adipose tissue and the rate of incorporation of carbon precursors into adipose tissues was greater in subcutaneous than in intramuscular adipose tissues. In comparison of amino acids, the rate was highest (p<0.05) with leucine followed by isoleucine and ${\alpha}$-ketoisocaproic acid in subcutaneous adipose tissue, in which there were no differences. Also, in intramuscular tissue, leucine was highest (p<0.05), and the rate of incorporation decreased in the same order. The rates of carbon precursor incorporation appeared to be higher in subcutaneous than in intramuscular tissue. For incorporation of radio-labelled acetate and glucose into intramuscular adipose tissue. preincubated for 48 hrs with insulin and IGF-1, insulin was the most effective to stimulate the incorporation of precursors in both substrates but there was no difference between insulin and IGF-1 in glucose incorporation. For glyceride-fatty acid synthesis, acetate was significantly (p<0.05) greater than glucose in both subcutaneous and intramuscular adipose tissue, and glyceride-glycerol synthesis was greater (p<0.05) for glucose than acetate in both adipose tissues. The rates of lipogenesis from both precursors were slightly greater in subcutaneous than intramuscular adipose tissue. There was significant (p<0.05) site effect in insulin treatment for glyceride-fatty acid synthesis. But there were no significance in control and norepinephrine. For glyceride-glycerol synthesis, there was no site effect caused by hormonal treatment. Insulin was the most effective (p<0.05) in glyceride fatty acid synthesis, while norepinephrine was the same as control. Compared with control, glyceride-glycerol synthesis from acetate in insulin treatment was significantly (p<0.05) low in subcutaneous, but high in intramuscular tissue. At the same time, in both tissues, it was lower in norepinephrine treatment than in control. Glyceride-glycerol synthesis from glucose was highest (p<0.05) in norepinephrine treatment followed by insulin although there was no significance between insulin and control. Lipolysis was not affected by insulin but was increased by norepinephrine when added to adipose tissue incubations in vitro. Rates of basal lipolysis were greater in subcutaneous adipose tissue than in intramuscular adipose tissue.

Quality characteristics of tangerine peel Soksungjang prepared from different koji strains (균주를 달리하여 제조한 귤피 속성장의 특성)

  • Youn, Young;Jeon, Sung-Hee;Yoo, Jeong-Hee;Jeong, Do-Yeon;Kim, Young-Soo
    • Food Science and Preservation
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    • v.23 no.1
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    • pp.117-126
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    • 2016
  • This study was performed to provide information for improving the quality characteristics of tangerine peel Soksungjang. Tangerine peel Soksungjangs were prepared with 10% tangerine peel powder by addition of different koji such as Rhizopus oligosporus (RS, RTS) and Aspergillus oryzae (AS, ATS). As a result, the moisture content was not changed and crude protein content decreased during fermentation in all samples. The pH of RS (Soksungjang manufactured with Rhizopus oligosporus koji) and RTS (Soksungjang manufactured with Rhizopus oligosporus koji and 10% tangerine peel powder) was not changed, whereas pH of AS (Soksungjang manufactured with Aspergillus oryzae) and ATS (Soksungjang manufactured with Aspergillus oryzae koji and 10% tangerine peel powder) decreased rapidly after 10 days of fermentation. The acidity values of all the samples increased during fermentation. The major free sugars were glucose, fructose, and maltose; while, the major organic acids were citric acid, succinic acid, acetic acid, and lactic acid in all the samples. The amino nitrogen content and protease activity were highest in the AS and ATS. Conversely, the reducing sugar content and ${\alpha}$-amylase activity were highest in the RS and RTS. The major free amino acids in all samples were glutamic acid, lysine, leucine, and arginine. Sixty kinds of volatile flavor compounds were identified from all samples; the majority of which were 21 alcohols, 14 hydrocarbons, 5 esters, and others identifies as acids, ketones, and aldehydes.

Processing and Physicochemical Properties of Collagen from Yellowfin Tuna (Thunnus albacares) Abdominal Skin (황다랑어 복부 껍질을 이용한 콜라겐의 제조 및 물리화학적 특성 해석)

  • Yoo, Sung-Jae;Cho, Sueng-Mock;Woo, Jin-Wook;Kim, Sang-Ho;Han, Yoo-Na;Ahn, Ju-Ryun;Kim, Su-Yeon;Kim, Tae-Wan;Kim, Seon-Bong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.41 no.6
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    • pp.427-434
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    • 2008
  • Processing of collagen from yellowfin tuna (Thunnus albacares) abdominal skins was optimized by response surface methodology and central composite design. The values of independent variables at optimal conditions were NaOH concentration: 0.5 N, NaOH treatment time: 36.2 hr, pepsin concentration: 1:4.9 ratio (0.245%, w/v), and digestion time: 48.1 hr, respectively. The collagen content estimated under optimal conditions was 33.1%, and the actual experimental collagen content was 32.3%. Physicochemical properties of collagen from yellowfin tuna abdominal skin were investigated by amino acids analysis, SDS-PAGE, FT-IR, viscosity and denaturation temperature. Amino acids content of the collagen was 21.0%. SDS-PAGE pattern of the collagen showed two different $\alpha$-chain (${\alpha}_1$- and ${\alpha}_2$- chain), $\beta$-component and $\gamma$-component. The spectrum of FT-IR of the collagen showed wavenumber at 3,434, 1,650, 1,542 and $1,235\;cm^{-1}$ representing the regions of amide A, I, II and III, respectively. Relative viscosity of the collagen decreased continuously on heating up to $32^{\circ}C$, and the rate of decrease was retarded in the temperature range of $35-50^{\circ}C$. Denaturation temperature (Td) of the collagen solution (0.06%, w/v) was $31^{\circ}C$ and was lower than calf skin collagen ($35^{\circ}C$).

Hypoallergenic and Physicochemical Properties of the A2 β-Casein Fractionof Goat Milk

  • Jung, Tae-Hwan;Hwang, Hyo-Jeong;Yun, Sung-Seob;Lee, Won-Jae;Kim, Jin-Wook;Ahn, Ji-Yun;Jeon, Woo-Min;Han, Kyoung-Sik
    • Food Science of Animal Resources
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    • v.37 no.6
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    • pp.940-947
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    • 2017
  • Goat milk has a protein composition similar to that of breast milk and contains abundant nutrients, but its use in functional foods is rather limited in comparison to milk from other sources. The aim of this study was to prepare a goat A2 ${\beta}$-casein fraction with improved digestibility and hypoallergenic properties. We investigated the optimal conditions for the separation of A2 ${\beta}$-casein fraction from goat milk by pH adjustment to pH 4.4 and treating the casein suspension with calcium chloride (0.05 M for 1 h at $25^{\circ}C$). Selective reduction of ${\beta}$- lactoglobulin and ${\alpha}_s$-casein was confirmed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis and reverse-phase high-performance liquid chromatography. The hypoallergenic property of A2 ${\beta}$-casein fraction was examined by measuring the release of histamine and tumor necrosis factor alpha from HMC-1 human mast cells exposed to different proteins, including A2 ${\beta}$-casein fraction. There was no significant difference in levels of both indicators between A2 ${\beta}$-casein treatment and the control (no protein treatment). The A2 ${\beta}$-casein fraction is abundant in essential amino acids, especially, branched-chain amino acids (leucine, valine, and isoleucine). The physicochemical properties of A2 ${\beta}$-casein fraction, including protein solubility and viscosity, are similar to those of bovine whole casein which is widely used as a protein source in various foods. Therefore, the goat A2 ${\beta}$-casein fraction may be useful as a food material with good digestibility and hypoallergenic properties for infants, the elderly, and people with metabolic disorders.

Modulation of G-CSF Secretion by Mutations of Non Alpha-Helical Region in N-Terminus (G-CSF 단백질 N-말단의 비 알파-Helix 영역의 돌연변이에 의한 분비 조절)

  • Park, Jeong-Hae;Park, Jung-Ae;Kang, Seok-Woo;Goo, Tae-Won;Chung, Kyung-Tae
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1778-1783
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    • 2011
  • Hematopoietic cytokines regulate production of blood cells by stimulating proliferation and differentiation of bone marrow cells. Among these hematopoietic cytokines, called hematopoitic growth factors, glranulocyte-colony stimulating Factor (G-CSF), which regulates growth of neutrophils, is one of important therapeutic factors because cancer patients suffer with neutropenia which is severe reduction of neutrophils after chemotherapy. Two groups of recombinant G-CSF have approved and used for therapeutic purposes and many researches are still on-going to produce recombinant G-CSF by different techniques. We engineered human G-CSF with Bombyx specific endoplasmic reticulum (ER) signal sequence, therefore, secretion of human G-CSF protein was improved in Bombyx mori-origined cell line, Bm5. The Bombyx ER signal sequence and human G-CSF matured protein region chimera was further remodeled at the N-terminus of matured G-CSF protein to understand roles of N-terminus on outer cellular secretion and/or production. Three different mutants were generated deleting three amino acids in non alpha-helical region in N-terminus in order to scan important amino acids for G-CSF secretion. One of 3 different N-terminal deletion mutants showed dramatically reduction of secreted amount of G-CSF indicating its important role on secretion. The data suggest that remodeling in non alpha-helical region of N-terminus is also important for recombinant G-CSF production.

Effects of Rice Bran, Flax Seed, and Sunflower Seed on Growth Performance, Carcass Characteristics, Fatty Acid Composition, Free Amino Acid and Peptide Contents, and Sensory Evaluations of Native Korean Cattle (Hanwoo)

  • Choi, Chang Bon;Kwon, Hana;Kim, Sung Il;Yang, Un Mok;Lee, Ju Hwan;Park, Eun Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.2
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    • pp.195-203
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    • 2016
  • This study was conducted to evaluate the effect of dietary supplementation with rice bran, flax seed, or sunflower seed to finishing native Korean cattle (Hanwoo) on growth performances, carcass characteristics, fatty acid composition, free amino acid and peptide contents, and sensory evaluations of Longissimus muscle (LM). A total of 39 Hanwoo steers (average age of 22.2 mo and average body weight (BW) of 552.2 kg) were randomly divided into Control, rice bran (RB), flax seed (FS), or Sunflower seed (SS) groups. The steers were group fed for 273 d until they reached an average age of 31.2 mo. Final BW was 768.2, 785.8, 786.2, and 789.0 kg, and average daily gain was 0.79, 0.85, 0.82, and 0.84 kg for the Control, RS, FS, and SS groups, respectively (p>0.05). Fat thickness of the FS group (19.8 mm) was greater (p<0.05) than that of the other groups. Final yield grade converted into numerical values was 2.0 for the RB group, 1.7 for the Control and SS groups, and 1.4 for the FS group. Marbling degrees for the Control, SS, RB, and FS groups were 5.3, 5.1, 4.7, and 4.6, respectively. Percentages of palmitic acid ($C_{16:0}$), stearic acid ($C_{18:0}$), and arachidic acid ($C_{20:0}$) in the LM were not different among the groups. Palmitoleic ($C_{16:1}$) acid was higher (p<0.05) in the SS group. The concentration of oleic acid was highest (p<0.05) in the Control group (47.73%). The level of linolenic acid ($C_{18:3}$) was 2.3 times higher (p<0.05) in the FS group compared to the other groups. Methionine concentration was (p<0.05) higher in FS (1.7 mg/100 g) and SS (1.2 mg/100 g) steers than in the Control or RB groups. Glutamic acid and ${\alpha}$-aminoadipic acid (${\alpha}$-AAA) contents were (p<0.05) higher in the FS group compared to the other groups. LM from the FS group had numerically higher (p>0.05) scores for flavor, umami, and overall palatability in sensory evaluations. In conclusion, supplementation of flax seed to diets of finishing Hanwoo steers improved sensory evaluations which might have been caused by increases in flavor related amino acids such as methionine, glutamic acid and ${\alpha}$-AAA and peptides, anserine and carnosine, and their complex reactions.

Rational Introduction of Disulfide Bond to Enhance Optimal Temperature of Lipomyces starkeyi $\alpha$-Dextranase Expressed in Pichia pastoris

  • Chen, Lin;Yu, Chao;Zhou, Xiangshan;Zhang, Yuanxing
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1506-1513
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    • 2009
  • $\alpha$-Dextranase, which can hydrolyze dextran, is largely used in the sugar industry. However, a thermostable $\alpha$-dextranase is needed to alleviate the viscosity of syrups and clean blocked machines. Thus, to improve the optimal temperature of Lipomyces starkeyi $\alpha$-dextranase expressed by Pichia pastoris, the rational introduction of a de novo designed disulfide bond was investigated. Based on the known structure of Penicillium minioluteum dextranase, L. starkeyi $\alpha$-dextranase was constructed using homology modeling. Four amino acids residues were then selected for site-directed mutagenesis to cysteine. When compared with the wild-type dextranase, the mutant DexM2 (D279C/S289C) showed a more than $13^{\circ}C$ improvement on its optimal temperature. DexM2 and DexM12 (T245C/N248C, D279C/S289C) also showed a better thermal stability than the wild-type dextranase. After the introduction of two disulfide bonds, the specific activity of DexM12 was evaluated and found to be two times higher than that of the wild-type. Moreover, DexM12 also showed the highest $V_{max}$.

Effects of Intrarenal Arterial Infusion of Pro-Atrial Natriuretic Peptides on Renal Function in Unanesthetized Rabbits (가토 신장기능에 미치는 Pro-Atrial Natriuretic Peptide의 영향)

  • Lee, Jeong-Eun;Cho, Kyung-Woo;Kim, Suhn-Hee
    • The Korean Journal of Physiology
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    • v.24 no.1
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    • pp.131-144
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    • 1990
  • It is well known that the atrial natriuretic peptide (ANP) has a prepro-hormone of 151 amino-acids which loses their hydrophobic signal peptide to form 126 amino acid prohormone. The whole prohormone is released and then cleaved by proteases into more than one circulating forms. Recently, Winters et al. (1988a, b) reported that high concentrations of N-terminal fragments of prepro-ANP $(26{\sim}55),\;(56{\sim}92)\;and\;(104{\sim}123)$ were detected in human plasma. However, their physiological roles have not been established. The present study was conducted to determine whether the N-terminal fragments of pro-ANP have any effect on the renal function and to compare the effect with those of G-terminal fragments of pro-ANP The results indicate that intrarenal arterial infusions of prepro-ANP $(26{\sim}41),\;(26{\sim}55),\;(56{\sim}92)\;and\;(104{\sim}123)$ induced no significant changes in renal function. Whereas ${\alpha}-human$ ANP $(prepro-ANP,\;124{\sim}151)$ and pro-ANP caused a significant increase in urine volume, renal plasma flow, glomerular filtration rate, urinary excretions of sodium, chloride and potassium, and fractional excretion of sodium. These results suggest that the N-terminal fragments of pro-ANP are ineffective, while the C-terminal fragments retain the natriuretic and diuretic activities.

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