• Title/Summary/Keyword: beta-Amino acids

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Characterization of the ${\beta}-Cyclodextrin$ Glucanotransferase Bacillus firmus var. alkalophilus and Its Expression in E. coli

  • Park, Tae-Hyung;Shin, Hyun-Dong;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.811-819
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    • 1999
  • The ${\beta}-CGTase$ gene of alkalophilic Bacillus firmus var. alkalophilus was cloned into E. coli using $pZErO^{TM}-2$ as a vector. The cloned gene encoded a total of 710 amino acid residues consisting of 674 amino acids of the matured protein and 36 amino acids of the signal peptide, including 20 amino acids from the lacZ gene in the vector. Although the cloned ${\beta}-CGTase$ gene did not contain the promoter and start codons, it was expressed by the lac promoter and lacZ start codon in the $pZErO^{TM}$ vector. A comparison was made with the amino acid sequence and ten other CGTases from Bacillus sp. Also, ten highly conserved regions, which are important amino acid residues in catalysis of CGTase, were identified. The lac promoter used for expression of the ${\beta}-CGTase$ gene was induced constitutively in recombinant E. coli even without IPTG possibly because of a lack of the lacI gene in both host and vector, repressing the lacZ gene in the lac operon. Its expression was catabolically repressed by glucose, however, its repression was reduced by soluble starch, mainly because of the extremely high increase of the cAMP level. ${\beta}-CGTase$ can be overproduced in the recombinant E. coli by maintaining intracellular cAMP levels mostly through the intermittent feeding of glucose during cultivation.

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Protein Context-Dependent Hydrophobicity of Amino Acids in Protein

  • Cho, Hanul;Ham, Sihyun
    • Proceeding of EDISON Challenge
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    • 2016.03a
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    • pp.163-166
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    • 2016
  • Hydrophobicity is the key concept to understand the water plays in protein folding, protein aggregation, and protein-protein interaction. Traditionally, the hydrophobicity of protein is defined based on the scales of the hydrophobicity of residue, assuming that the hydrophobicity of free amino acids is maintained. Here, we explore how the hydrophobicity of constituting amino acids in protein rely on the protein context, in particular, on the total charge and secondary structures of a protein. To this end, we calculate and investigate the hydration free energy of three short proteins based on the integral-equation theory of liquids. We find that the hydration free energy of charged amino acids is significantly affected by the protein total charge and exhibits contrasting behavior depending on the protein total charge being positive or negative. We also observe that amino acids in the ${\beta}-sheets$ display more enhanced the hydrophobicity than amino acids in the loop, whereas those in the ${\alpha}-helix$ do not clearly show such a tendency. And the salt-bridge forming amino acids also exhibit increase of the hydrophobicity than that with no salt bridge. Our results provide novel insights into the hydrophobicity of amino acids, and will be valuable for rationalizing and predicting the strength of water-mediated interaction involved in the biological activity of proteins.

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Cloning of $\beta$-Tubulin Gene and Effect of Pencycuron on Tubulin Assembly in Rhizoctonia solani

  • Roh, Seong-Hwan;Heung-Tae KIm;Isamu-Yamaguchi
    • The Plant Pathology Journal
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    • v.15 no.1
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    • pp.68-71
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    • 1999
  • To illustrate the action mechanism of pencycuron on Rhizoctonia solani, two experiments were conducted including the comparison of amino acids of $\beta$-tubulin between R-C (sensitive isolate) and Rh-131 (non-sensitive isolate), and the inhibitory effect of pencycuron on tubulin assembly in vitro. Both $\beta$-tubulin genes of R-C and Rh-131 proved to have 1,582 nucleotides encoding a protein of 445 amino acids, showing 98% homology in amino acid sequences between them. It was found that codons at 103, 236, and 267 for lysine (AGG), valine (GTC) and isoleucine (ATT) in R-C were replaced by codons for methionine (ATG), isoleucine (ATT) and methionine (ATG) in Rh-131, respectively. No inhibitory effect of pencycuron on the tubulin assembly was observed. It suggests that pencycuron may have no direct inhibitory effects on the assembly of tubulin at least in vitro.

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Effect of addition amino acids on the mycelial growth and the contents of β-glucan and γ-aminobutyric acid (GABA) in Sparassis latifolia (아미노산 첨가가 꽃송이버섯 균사체 성장 및 베타글루칸, GABA 함량 변화에 미치는 영향)

  • Jo, Han-Gyo;Shin, Hyun-Jae
    • Journal of Mushroom
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    • v.15 no.1
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    • pp.38-44
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    • 2017
  • Sparassis latifolia (formerly S. crispa) is used in food and nutraceuticals or dietary supplements, as rich in flavor compounds and ${\beta}-glucan$. Some previous studies have reported the effects of mushroom on brain function, including its neuroprotective effect. Thus, for this mushroom to be used as an effective nutraceutical for brain function, it would be desirable for it to contain other compounds such as ${\gamma}-aminobutyric$ acid (GABA) in addition to ${\beta}-glucan$. In this study, the enhancement of growth and GABA production in the mycelium of medicinal and edible mushroom S. latifolia was investigated. Amino acids were added externally as the main source of nutrition, and the effects of amino acids were investigated using liquid medium, specifically amino acid-free potato dextrose broth (PDB). The amino acids added were L-glutamic acid (named PDBG medium) and L-ornithine (named PDBO medium). The growth of mycelia was determined to be $0.9{\pm}0.00g/L$, $2.2{\pm}0.16g/L$, and $1.93{\pm}0.34g/L$ PDBG respectively. The GABA content was $21.3{\pm}0.9mg/100g$ in PDB medium, and it in PDBG 1.4% medium, at $115.4{\pm}30.2mg/100g$. However, the PDBO medium was not effective in increasing the GABA content of mycelia. Amino acids had little effect on the ${\beta}-glucan$ content of mycelia. The ${\beta}-glucan$ content was $39.7{\pm}1.4mg/100mg$, $34.4{\pm}0.2mg/100mg$, and $35.2{\pm}9.2mg/100mg$ in PDB, PDBG 1.8% and PDBO 1.4% media, respectively. Addition of glutamic acid and ornithine positively affected the growth of S. latifolia mycelia, and glutamic acid positively affected GABA production; no degradation of GABA was observed with addition of glutamic acid.

Studies on the Snake Wines(Part 1) On the Free amino acid (한국산(韓國産) 사주(蛇酒)에 관(關)한 연구(硏究)(제(第)1보(報)) 사주(蛇酒)의 Free Amino Acid 에 관(關)하여)

  • Park,, Yoon-Choong;Chung,, Soon-Lyang
    • Korean Journal of Food Science and Technology
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    • v.1 no.1
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    • pp.74-77
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    • 1969
  • In this study, three kinds of Korean snake wine (50 V% alcohol extracts) were determined by amino acid analyzer and were discussed as follows. 1. Salmosa Ju and Nungsa Ju were composed of 22 free amino acids: cysteic acid, aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, cystine, valine, methionine, iso-leucine, allo-iso-leucine, leucine, nor-leucine, tyrosine, phenylalanine, ${\beta}-alanine$, lysine, arginine, histidine, trytophan. 2. Doksa Ju were composed of 21 free amino acids which were all same as above except missing histidine. 3. The free amino acid composition were almost identical in Doksa Ju, Salmosa Ju and Nungsa Ju quantitatively. 4. The contents of cysteic acid, glutamic acid, glycine, alanine, valine, leucine and lysine were relatively high, on the other hand, methionine, allo-iso-leucine, nor-leucine, and tryptophan were trace amount in every snake wine. 5. Eleven unknowns of ninhydrin positives were identified in the every snake wine. 6. The free amino acids in snake wines were various in kind as compared with in beer, Japanese sake and Korean Tack Ju. Especially cysteic acid, allo-iso-leucine, nor-leucine and ${\beta}-alanine$ in snake wines were missed in every cereal wine.

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Studies on the Composition of Amino Acid in Retina and Lens Body of the Frogs (개구리 망막체와 수정체의 Amino 산 성분에 관한 연구)

  • 강성호
    • The Korean Journal of Zoology
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    • v.2 no.2
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    • pp.10-14
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    • 1959
  • The retinae and the lens bodies of the frogs were hydrolyzed with 20% hydrochloric acid, and their amino acids were separated by paper chromatography. As a result of it the following were confirmed : (1) The retinae and the lens bodies were the same incomposition, and aspartic acid, glutamic acid, serine ,tyrosine, glycine, lysine arginine, threonine, alanine, histidine, proline, methionine, valine , phenylalanine, leucine, and two unknown substances were separated. (2) The free amino acids in the retinae were extracted with 80% ethyl alcohol and then separated by paper chromatography. Though their separation was not so definite , serine, gultaminc acid, and glycine were always separated regardless of the amount of the sample. When the amount of the smaple was enough , $\beta$-alanine , ${\gamma}$-amino butyric acid and methionine +valine were also separated. (2) The free amino acids in the retinae were extracted with 80% ethyl alcohol and then separated by paper chromatography. Though their separation was not so definite, serine, glutamic acid, and glycine were always separated regardless of the amount of the sample . When the amount of the sample was enough, $\beta$-alanine , ${\gamma}$-amino butyric acid and methionine + valine were also separated.

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Physicochemical Properties of Domestic Cherry Tomato Varieties (국내산 방울토마토의 이화학적 특성)

  • Ahn, Jun-Bae
    • Culinary science and hospitality research
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    • v.23 no.7
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    • pp.42-49
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    • 2017
  • This study was conducted to determine the nutritional value of domestic cherry tomato varieties (Summerking, Qutiquti, and Minichal). The levels of amino acids, amino acid derivatives, and ${\gamma}-aminobutyric-acid$ (GABA) were analyzed using ion chromatography. In domestic cherry tomatoes, eighteen free amino acids were found including L-glutamic acid (L-Glu), L-glutamine (L-Gln), and L-aspartic acid (L-Asp). L-Glu was the most abundant amino acid, ranging from 1,533.17 mg/100 g to 1,920.65 mg/100 g (dry weight). The next abundant amino acids were L-Gln, ranging from 784.68 mg/100 g to 1,164.36 mg/100 g and L-Asp, ranging from 320.73 mg/100 g to 387.22 mg/100 g. Domestic cherry tomatoes contained eight essential amino acids except tryptophan and the total essential amino acid content was 297.30~432.43 mg/100 g (dry weight), which was 8.92~10.61% of total free amino acid. Several amino acid derivatives were found: L-carnitine (L-Car), hydroxylysine (Hyl), o-phosphoethanolamine (o-Pea), phosphoserine (p-Ser), ${\beta}-alanine$ (${\beta}-Ala$), N-methyl-histidine (Me-His), ethanolamine ($EtNH_2$), and L-citrulline (L-Cit). L-Car, transporting long-chain fatty acid into mitocondrial matrix, was the most abundant amino acid derivative in all domestic cherry tomatoes. A high level of GABA (313.18~638.57 mg/100 g), known as a neurotransmitter, was also found in all three domestic cherry tomatoes. These results revealed that domestic cherry tomatoes have a good balance of nutrient and bioactive compounds. Therefore, cherry tomatoes can be used as a functional food material.

COOH-Terminal Animo Acids of Tethered-Buman Glycoprotein Bormone $\alpha$-Subunit Play an Important Role for Secretion

  • Min, K.S;Yoon, J.K.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.395-399
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    • 2002
  • Human chorionic gonadotropin (hCG) is a member of the glycoprotein hormone family which includes FSH. hCG TSH. These hormone family is characterized by a heterodimeric structure composed a common $\alpha$-subunit noncovalently linked to a hormone specific $\beta$-subunit. To determine u and $\beta$ -subunits can be synthesized as a single polypeptide chain (tethered-hCG) and also display biological activity, the tethered-hCC and -FSH molecule by fusing the carboxyl terminus of the hCG $\beta$-subunit to the amino terminus of the $\alpha$-subunit was constructed. To determine the importance of $\alpha$ COOH -terminal amino acid, we also deleted the $\alpha$ COOH-terminal amino acids. The expressing vectors were transfected into CHO-K 1 cells. The tethered-wthCG and -wtFSH was efficiently secreted. The $\alpha$ Δ83hCG and $\alpha$ Δ 83FSH mutants had no secretion. These results are the first conclusive evidence that COOH-terminal amino acids are very important for secretion in human glycoprotein hormone $\alpha$-subunit. These results demonstrated that the $\alpha$ Δ83hCG and $\alpha$ Δ 83FSH mutants could be play a pivotal role in the secretion of tethered-molecule.

Stereoselective Synthesis of Diverse α-Hydroxy-β-amino Acids and It's Application for Synthesis of Dipeptide Expecting as a Protease Inhibitor

  • Jang, Sang-Hun;Kim, Ju-Young;Kim, Min-Kyu;Han, Jeong-Woo;Park, Ki-Hun;Yoon, Yong-Jin;Lee, Sang-Gyeong
    • Bulletin of the Korean Chemical Society
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    • v.30 no.1
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    • pp.163-171
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    • 2009
  • Few $\alpha$-Hydroxy-$\beta$-amino acids were synthesized via various nucleophilic addition of the epoxide and followed by stereoselective nucleophilic substitution reaction and eliminative cleavage of the acetal selectively in diacetal compound. One of the synthesized $\alpha$-Hydroxy-$\beta$-amino acid reacted with L-leucine methylester to give corresponding dipeptide in good yields.

Temperature, Medium and Structural Effects on the Acid Dissociation Constants of Certain Schiff Bases Derived from Isatin with Some Amino Acids and Aroylhydrazines

  • Hassaan, Aly M.A.;Quenawy, M.T.A.
    • Archives of Pharmacal Research
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    • v.16 no.3
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    • pp.180-185
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    • 1993
  • The acid dissociation constants of certain Schiff bases derived from isatin with p-substituted benzoyl hydrazines and somee amino acids of glycine, $\beta$-and $\alpha$-alanine, valine, $\beta$-phenyl-$\alpha$-alanine and anthranilic acid have been detemined potentiometrically at different temperatures in different aquo-organic solvent mixtures (ethnol, dioxane, dimethyl formamide, methanol, acetone and tetrahydrofurance). The pKa values were demonstrated on the light of the different electornic and steric effects of the substituents and the sovlent characteristics. In all the mixed media used, ionization of the compounds decreased by increasing the mole fraction of the organic cosolvent. Theemodynamic parameters $(\Delta{H}^\circ,\;\Delta{G}^\circ,\;\Delta{S}^\circ)$ were evaluated. The structural effects of the investigated compounds on these parameters were reported and discussed.

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