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

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Cloning and Expression of a Paenibacillus sp. Neopullulanase Gene in Saccharomyces cerevisiae Producing Schwanniomyces occidentalis Glucoamylase

  • Kim, Hyo-Jeong;Park, Jeong-Nam;Kim, Hee-Ok;Shin, Dong-Jun;Chin, Jong-Eon;Blaise Lee, Hwang-Hee;Chun, Soon-Bai;Bai, Suk
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.340-344
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    • 2002
  • A gene, npl, encoding neopullulanase from Paenibacillus sp. KCTC 8848P was cloned and expressed in Escherichia coli. It consisted of an open reading frame of 1,530 bp for a protein that consisted of 510 amino acids with a molecular weight of 58,075 Da. The deduced amino acid sequence of the neopullulanase gene had $92\%$ identity with the neopullulanase of Bacillus polymyxa. The npl gene was also expressed in Saccharomyces cerevisiae secreting Schwanniomyces occidentalis glucoamylase (GAM1) under the control of the yeast actin gene (ACT1) promoter. Secretion of the neopullulanase was directed by the yeast mating pheromone ${\alpha}$ -factor ($MF{\alpha}1$) prepro region. Enzyme assays confirmed that co-expression of npl and GAM1 enhanced starch and pullulan degradation by S. cerevisiae.

Comparison of Protein Binding Polysaccharide from Agaricus blazei Murill Prepared by Ultrafiltration and Spray-Drying Process

  • Hong, Joo-Heon;Choi, Yong-Hee;Youn, Kwang-Sup
    • Journal of Applied Biological Chemistry
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    • v.50 no.1
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    • pp.1-5
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    • 2007
  • Chemical properties of spray-dried powders separated based on molecular weight from crude protein binding polysaccharide (CP-SD) of Agraricus blazei were examined. Contents of ${\beta}$-glucan in SD-1, SD-2 and SD-3 were 18.67%, 48.24%, and 37.15% respectively, and SD-2 (10-150 kDa) showed the highest molecular weight. Obtained ${\beta}$-glucans were not pure glucan, but was determined to be an acidic proteo-heteroglycan with a large amount of glucose (74.46-80.05%), galactose (8.91-15.2%), and mannose (4.9-5.46%). Composition of their amino acids was mainly aspartic and glutamic acids. FT-IR spectrum revealed SD-1, SD-2 and SD-3 were structures of ${\beta}$-1,3-glucans and ${\alpha}$-1,6-glucans at 890 and 930 $cm^{-1}$, respectively, signals of ${\alpha}$-1,6-glucans for CP-SD was not found. Useful CP-SD was recovered from A. blazei for preparation of three powder types as food materials.

Nutritional Compositions of Three Traditional Actinidia (Actinidia arguta) Cultivars Improved in Korea (국내에서 개량된 3가지 토종 다래 품종의 영양성분 분석)

  • Jin, Dong Eun;Park, Seon Kyeong;Park, Chang Hyeon;Seung, Tae Wan;Heo, Ho Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.12
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    • pp.1942-1947
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    • 2014
  • Nutritional compositions of Korean traditional actinidia (Actinidia arguta) cultivars, such as Otumsense, Chiak, and Skinny green, were investigated as high value-added food substances. Among minerals, K content of three cultivars was the highest, and P, Ca, and Na contents were relatively higher than those of other minerals. Contents of essential amino acids were analyzed as follows: Otumsense (198.48 mg/100 g), Chiak (413.50 mg/100 g), and Skinny green (270.07 mg/100 g). Total amino acids of Chiak were the highest among the three cultivars, and major amino acids of the three cultivars were glutamic acid and aspartic acid. Analysis of fatty acids showed that major fatty acids were palmitic acid as a saturated fatted acid and ${\alpha}$-linoleic acid as an unsaturated fatty acid in the three cultivars. Glucose and fructose were major free sugar constituents in the three cultivars. Total free sugar content of Otumsense was relatively higher than others. Finally, niacin and vitamin $B_6$ of Skinny green (1.55 mg/100 g, 1.92 mg/100 g) were the highest among the cultivars. However, ${\beta}$-carotene as a vitamin A precursor ($3.82{\mu}g/100g$) and vitamin C as a natural antioxidant substance (47.18 mg/100 g) of Otumsense cultivar were the highest.

Transport of Sulfanilic Acid via Microbial Dipeptide Transport System

  • Hwang, Se-Young;Ki, Mi-Ran;Cho, Suk-Young;Yoo, Ick-Dong
    • Journal of Microbiology and Biotechnology
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    • v.5 no.6
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    • pp.315-318
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    • 1995
  • Sulfanilic acid (4-aminobenzenesulfonic acid) alone is normally not permeant in bacteria but can be readily delivered via the microbial dipeptide transport system. A dipeptidyl derivative of this compound, L-phenylalanyl-L-2-sulfanilylglycine (PSG), prepared by attachment of its primary amino group to the phenylalanyl $\alpha$-glycine moiety, appeared to have a Km of 0.125 mM and a Vmax of 1.9 nmoles/ml/min ($A_{660}$, 1.0) in Escherichia coli. From competitive spectrophotometric analysis, it was found that the type of amino acids in both of the N- and C-terminals affected the kinetic power of dipeptides. The growth inhibitory effect of PSG was over 7 times more potent than that of the sulfanilic acid against E. coli, suggesting that this potential inhibition was presumably due to the increased hydrophobic nature of the sulfanilyl dipeptide.

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The Quality Characteristics of Sauce Made with Rock Lobster (바다가재를 이용한 소스의 품질 특성)

  • 이경임
    • The Korean Journal of Community Living Science
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    • v.15 no.3
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    • pp.15-20
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    • 2004
  • The purpose of this study was to evaluate the sensory characteristics and investigate the nutrients composition of sauce made with rock lobster shells or heads. Rock lobster head sauce(LHS) turned out to have better appearance, color, taste, flavor, texture and viscosity than rock lobster shell sauce(LSS). The crude protein content was 1.95% in LHS and 1.56% in LSS, and the crude lipid content was 1.60% in LHS and 2.27% in LSS. The major free amino acids of LHS were urea, a-aminoadipic acid, L-alanine, taurine, and those of LSS were urea, $\alpha$-aminoadipic acid, $nu$- and DL-$\beta$-aminobutyric acid. The major free fatty acids of both samples were palmitoleic acid and oleic acid, with a high quanitity of palmitoleic acid in LHS, and a significant amount of linoleic acid in LSS. The major minerals in rock lobster sauce were potassium and sodium.

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Mainchain NMR Assignments and secondary structure prediction of the C-terminal domain of BldD, a developmental transcriptional regulator from Streptomyces coelicolor A3(2)

  • Kim, Jeong-Mok;Won, Hyung-Sik;Kang, Sa-Ouk
    • Journal of the Korean Magnetic Resonance Society
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    • v.17 no.1
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    • pp.59-66
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    • 2013
  • BldD, a developmental transcription factor from Streptomyces coelicolor, is a homodimeric, DNA-binding protein with 167 amino acids in each subunit. Each monomer consists of two structurally distinct domains, the N-terminal domain (BldD-NTD) responsible for DNA-binding and dimerization and the C-terminal domain (BldD-CTD). In contrast to the BldD-NTD, of which crystal structure has been solved, the BldD-CTD has been characterized neither in structure nor in function. Thus, in terms of structural genomics, structural study of the BldD-CTD has been conducted in solution, and in the present work, mainchain NMR assignments of the recombinant BldD-CTD (residues 80-167 of BldD) could be achieved by a series of heteronuclear multidimensional NMR experiments on a [$^{13}C/^{15}N$]-enriched protein sample. Finally, the secondary structure prediction by CSI and TALOS+ analysis using the assigned chemical shifts data identified a ${\beta}-{\alpha}-{\alpha}-{\beta}-{\alpha}-{\alpha}-{\alpha}$ topology of the domain. The results will provide the most fundamental data for more detailed approach to the atomic structure of the BldD-CTD, which would be essential for entire understanding of the molecular function of BldD.

Studies on the Compositional Change of Composts During Mushroom Cultivation (양송이 재배(栽培)에 따른 재배상퇴비(栽培床堆肥)의 성분변화(成分變化)에 관(關)한 연구(硏究))

  • Namgung, Hee
    • Applied Biological Chemistry
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    • v.18 no.4
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    • pp.203-218
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    • 1975
  • In order to investigate the compositional change oil composts during the growing of cultivated mushroom (Agaricus bisporus), composts and mushrooms during the period of filling to ending under commercial conditions were subjected to chemical analyses. The results are summarized as follows and the mechanism of composting for mushroom cultivation was proposed. 1) The temperature change of growing bed and room was observed and the yield of mushroom for each cropping time was recorded to get $15.6kg/m^2$ in total crops. 2) Composts after filling showed pH 8.2 which dropped to 6.4 after casing and continued so up to ending. 3) On the dry weight basis of composts, crude ash increased whereas total nitrogen, ether extract and crude fibre decreased gradually to bring about the lowering of organic matter. 4) Total nitrogen of composts decreased gradually and more insoluble nitrogen was lost than soluble nitrogen. The C/N ratio of composts was initially 21 which was gradually lowered to 16. 5) The losses of ${\alpha}-cellulose$, pentosan and lignin in composts were 87%, 75%, and 60%, respectively, in which ${\alpha}-cellulose$ decreased markedly after casing. 6) Free reducing sugars of composts increased continuously. Gradually increased free amino acids till second cropping decreased again thereafter. Composts at the filling stage contained alanine, glutamic acid, glycine and serine in which glycine decreased markedly whereas proline increased remarkably upon mushroom cultivation. 7) Among minerals of composts, phosphorus and zinc tended to decrease, potassium and copper tended to increase anti sodium showed no marked change. 8) In comparison of mushrooms from different cropping time with respect to proximate composition, minerals, free reducing sugars and amino acids, no marked difference was observed. However, a little higher values were observed in crude fat, free reducing sugars and sodium content for early crops and in free amino acids and phosphorus content for late crops. Twelve free amino acids including alanine, serine, threonine, and glutamic acid were detected in the cultivated mushroom. 9) According to above experimental results, it was possible to support the mechanism of compositing that the formation of ammonia and decomposition of carbohydrates by mesophiles are followed by protein biosynthesis, formation of microbial bodies and nitrogen-rich lignin humus complex by thermophiles, thus supplying necessary nutrients for mushroom growth, along with residual carbohydrates.

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Detection of Chemical Characteristics in Hamcho (Salicornia herbacea L.) according to Harvest Periods (함초(Salicornia herbacea L.)의 채취 시기별 이화학적 특성 탐색)

  • Cha, Jae-Young;Jeong, Jae-Jun;Kim, Yong-Taek;Seo, Won-Seok;Yang, Hyun-Ju;Kim, Jin-Sook;Lee, Yong-Soo
    • Journal of Life Science
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    • v.16 no.4
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    • pp.683-690
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    • 2006
  • The chemical compositions of amino acids, minerals, betaine, and polyphenolic compounds from Salicornia herbacea (Hamcho) according to harvest periods were analyzed. Changes of chemical characteristics in water and ethanol extracts from Hamcho were evaluated for titratable acidity, pH, soluble solid, and Hunter's color values. The antioxidative activity of water extract from Hamcho was also determined by DPPH $({\alpha},{\alpha}'-diphenyl-{\beta}-picrylhydrazyl)$ scavenging radical activity. Total polyphenolic compounds of Hamcho were shown the highest in August harvested by 201.6 ppm. The betaine of Hamcho water-extract was identified by high performance liquid chromatography (HPLC), which content was 0.248%, 0.269% and 0.204% in June, August, and October, respectively. Major compositional amino acids (mg%) were glutamic acid (582: 519: 664), proline (552: 471: 322), phenylalanine (480: 431: 424), aspartic acid (322: 297: 330), and arginine (282: 321: 483) in June, August, and October, respectively, and major free amino acids (mg%) were proline (9.7: 3.4), asparagine (6.7: 1.4), hydroxyproline (6.4: 2.8), valine (3.9: 2.5), arginine (1.7: 3.0) in June and August, respectively. Mineral contents (mg%) were Na (5,695: 7,536: 5,529), K (1,640: 963: 931), Mg (359: 428: 348), Ca (221: 234: 251), and P (207: 189: 259) in June, August, and October, respectively. Especially, K was high in June, Na and Mg were high in August, and Ca and P were high in October, respectively. DPPH scavenging radical activity was shown in the following order; 0.05% butylated hydroxytoluene (BHT)> August> June> October harvested Hamcho. The chemical components of polyphenolic compounds, betaine, amino acids and minerals were changed by harvest periods according to the growing season, and the highest concentrations of polyphenolic compounds and betaine of Hamcho were shown in August harvested.

Effect of Dietary Taurine or Glycine Supplementation on Plasma and Liver Free Amino Acid Concentrations in Rats (식이내의 타우린 또는 글라이신 보강이 흰쥐의 혈장과 간의 유리아미노산 농도 및 패턴에 미치는 영향)

  • 박정은
    • Journal of Nutrition and Health
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    • v.31 no.2
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    • pp.126-134
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    • 1998
  • Our previous study demonstrated that dietary taurine or glycine supplementation significantly lowered plasma and hepatic cholesterol and triglyceride concentrations in rats fed a cholesterol-free diet. In the present study, the effect of long term dietary taurine or glycine supplementation, for the purpose of preventing and/or treating of hyperlipidemia and other known biological functions, on plasma and hepatic free amino acid concentrations and profiles were evaluated in rats. Three groups of male rats(110-130g) were fed a control diet(CD), taurine-supplemented diets(TSD ; CD+ 1.5% taurine) or glycine-supplemented diet(GSD ; CD+1.5% glycine) for 5 weeks. Plasma and hepatic free amino acid concentrations were determined by an automated amino acid analyzer based on ion-exchange chormatography. The feeding of TSD for 5 weeks yielded a 444% higher plasma taurine concentration , and the feeding GSD for the same period resulted in a 143% higher plasma glycine level in rats compared to those fed DB. Hepatic taurine concentration was significantly higher in rats fed TSD(145% increase) compared to the control rats. However, hepatic glycine concentration was not influenced by dietary glycine supplementation , which implies that the massive dose of glycine entering the body was more rapidly metabolized or excreted than taurein. Dietary taurine or glycine supplementation resulted in similar changes in plasma free amino acid concentrations, except in levels of taurine and glycine. Plasma levels of histidine, lysine, phenylalanine , alanine, proline, hydroxypoline, $\alpha$-aminogutyric acid, cystathionine and ethanolamine were significantly higher in rats fed TSD or GSD than those fed GD. Glycine supplementation did not change hepatic free amino acid concentrations as compared to CD. Concentrations of most hepatic free amino acids were not influenced by dietary taurine supplementation with the exception of significantly higher levels of asparate and tyrosine(56-63% increase) and lower levels of histidine and glutamate(33-34% decrease) compared to the control rats. These results suggest long-term dietary taurine or glycine supplementation resulted in increases in most plasma free amino acid levels, but did not cause a characteristic change in plasma aminogram pattern compared to rats fed CD.

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Metabolic Characteristic of the Liver of Dairy Cows during Ketosis Based on Comparative Proteomics

  • Xu, Chuang;Wang, Zhe;Liu, Guowen;Li, Xiaobing;Xie, Guanghong;Xia, Cheng;Zhang, Hong You
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.7
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    • pp.1003-1010
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    • 2008
  • The objective of the present study was to identify differences in the expression levels of liver proteins between healthy and ketotic cows, establish a liver metabolic interrelationship of ketosis and elucidate the metabolic characteristics of the liver during ketosis. Liver samples from 8 healthy multiparous Hostein cows and 8 ketotic cows were pooled by health status and the proteins were separated by two-dimensional-electrophoresis (2D-E). Statistical analysis of gels was performed using PDQuest software 8.0. The differences in the expression levels of liver proteins (p<0.05) between ketotic and healthy cows were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF-TOF) tandem mass spectrometry. Five enzymes/proteins were identified as being differentially expressed in the livers of ketotic cows: expression of 3-hydroxyacyl-CoA dehydrogenase type-2 (HCDH), acetyl-coenzyme A acetyltransferase 2 (ACAT) and elongation factor Tu (EF-Tu) were down-regulated, whereas that of alpha-enolase and creatine kinase were up-regulated. On the basis of this evidence, it could be presumed that the decreased expression of HCDH, which is caused by high concentrations of acetyl-CoA in hepatic cells, in the livers of ketotic cows, implies reduced fatty acid ??oxidation. The resultant high concentrations of acetyl-CoA and acetoacetyl CoA would depress the level of ACAT and generate more ??hydroxybutyric acid; high concentrations of acetyl-CoA would also accelerate the Krebs Cycle and produce more ATP, which is stored as phosphocreatine, as a consequence of increased expression of creatine kinase. The low expression level of elongation factor Tu in the livers of ketotic cows indicates decreased levels of protein synthesis due to the limited availability of amino acids, because the most glucogenic amino acids sustain the glyconeogenesis pathway; thus increasing the level of alpha-enolase. Decreased protein synthesis also promotes the conversion of amino acids to oxaloacetate, which drives the Krebs Cycle under conditions of high levels of acetyl-CoA. It is concluded that the livers of ketotic cows possess high concentrations of acetyl-CoA, which through negative feedback inhibited fatty acid oxidation; show decreased fatty acid oxidation, ketogenesis and protein synthesis; and increased gluconeogenesis and energy production.