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

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Stereospecific Synthesis of the (2R,3S)- and (2R,3R)-3-Amino-2-hydroxy-4-phenylbutanoic Acids from D-Glucono-δ-lactone

  • Lee, Jin Hwan;Kim, Jin Hyo;Lee, Byong Won;Seo, Woo Duck;Yang, Min Suk;Park, Ki Hun
    • Bulletin of the Korean Chemical Society
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    • v.27 no.8
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    • pp.1211-1218
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    • 2006
  • The enantiomerically pure (2R,3S)- and (2R,3R)-3-amino-2-hydroxy-4-phenylbutanoic acids (AHPBA) 1 and 3 are readily obtained from D-glucono-a-lactone. Both AHPBAs are the structural key units of KMI derivatives which are the potent inhibitors of BACE 1 ($\beta$-secretase) and HIV protease. Additionally, the obtained AHPBAs 1 and 3 are converted to dipeptides of bestatin stereoisomers 2 and 4.

Studies on the Removal of Protein Soils ( I ) -Characterization of Human Epidermal Stratum Corneum as Model Soils for Detergency Test- (단백질 오염의 세척거동에 관한 연구(I) -세척 시험용 모델 오염으로서의 인체 표피 각질층의 특성-)

  • Lee Jeong Sook;Kim Sung Reon
    • Journal of the Korean Society of Clothing and Textiles
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    • v.10 no.3
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    • pp.1-8
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    • 1986
  • The purpose of this study was to investigate the characteristics of human epidermal stratum corneum as protein model soils for detergency test. The stratum corneum was collected by scraping of the skin and purified with solvent. The results obtained were as follows: 1. Purified stratum corneum contained $92.38\%$ of crude protein. 2. In the amino acid compositions, contents of glycine, glutamic acid and serine were high and methionine and cystine were low. They were similar to fibrous $\alpha$-keratin consisted of stratum corneum. Whereas the content of polar amino acids was decreased, that of nonpolar amino acids was increased after enzyme hydrolysis. 3. The hydrolysis of stratum corneum with enzyme increased muck at initial reaction time and levelled off in 4$\~$6 hours. The hydrolysis with enzyme was improved effectively at its optimum temperature and pH. 4. The hydrolysis of stratum corneum with enzyme increased by the addition of surfactants. The order of compatibility with enzyme was in the order of Triton X-100>AOS>LAS.

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Effect of Thermal Pretreatment on the Functional Constituents of Waxy Corn (Zea mays L.)

  • Kim, Eun-Ok;Yu, Myong-Hwa;Lee, Ki-Teak;Yun, Kyoung-Seob;Choi, Sang-Won
    • Food Science and Biotechnology
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    • v.18 no.6
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    • pp.1336-1341
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    • 2009
  • Quantitative changes in functional constituents of waxy corn (Zea mays L.) by 5 different thermal pretreatments, including roasting, steaming, microwave, puffing, and extruding, were determined and compared with those of the raw waxy corn. There were no significant differences in fatty acid compositions among the corn treated with 5 thermal treatments. Levels of $\alpha$- and $\gamma$-tocopherols, soluble amino acids, and phytosterols decreased by thermal treatments, while those of p-coumaric and ferulic acids considerably increased by thermal treatments. In particular, the contents of tocopherols and phytosterols, and souble amino acid decreased significant in the steaming and puffing processes, respectively, whereas those of 2 free cinnamic acids increased significantly in the steaming and puffing processes. The extruding process showed smaller quantitative changes in tocopherols, phytosterols, and hydroxycinnamic acid derivatives compared to other heat pretreatments. These results suggest that the extruding process have a positive effect on valuable phytochemicals in waxy corn.

The Effect of synthetic Antioxidants on the Proteolytic enymes 1. The Effect of synthetic Antioxidants on the Activity of the $\alpha$-Chymotrypsin and Trypsin (합성 항산화제가 단백질 분해효소에 미치는 영양 -제1보,$\alpha$ -Chymotrypsin 과 trypsin의 활성에 미치는 영양-)

  • 김상옥
    • Journal of the Korean Home Economics Association
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    • v.19 no.2
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    • pp.183-188
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    • 1981
  • This study was conducted to investigate the effects of synthetic antioxidants con the degradation of angiotensin II which is made up of 8 amino acids: Asp-Arg-Val-Try-Ile-Gly-Pro-Phe, by the $\alpha$-chymotrypsin and trypsin. the results obtained were as follow; 1. Dibutyl hydroxytoluene, butyl hydroxyanisole and sodium L-ascorbate showed no inhibitory effect on the activity of $\alpha$-chymotrypsin on the angiotensin II, but ethyl protocathechuate inhibited. its activity at the concentration of 100ppm. However, the angiotension II was gradually degradated by $\alpha$-chymotrypsin after one hour incubation with ethylprotecathechuate. 2. Butyl hydroxyanisole inhibited trypsin activities above 100ppm, but no inhibitory activities was observed by the other antioxidants used in this experiment.

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Cloning and Expression of a Thermostable ${\alpha}$-Galactosidase from the Thermophilic Fungus Talaromyces emersonii in the Methylotrophic Yeast Pichia pastoris

  • Simila, Janika;Gernig, Anita;Murray, Patrick;Fernandes, Sara;Tuohy, Maria G.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1653-1663
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    • 2010
  • The first gene (${\alpha}$-gal1) encoding an extracellular ${\alpha}$-Dgalactosidase from the thermophilic fungus Talaromyces emersonii was cloned and characterized. The ${\alpha}$-gal1 gene consisted of an open reading frame of 1,792 base pairs interrupted by six introns that encoded a mature protein of 452 amino acids, including a 24 amino acid secretory signal sequence. The translated protein had highest identity with other fungal ${\alpha}$-galactosidases belonging to glycosyl hydrolase family 27. The ${\alpha}$-gal1 gene was overexpressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris. Recombinant ${\alpha}$-Gal1 was secreted into the culture medium as a monomeric glycoprotein with a maximal yield of 10.75 mg/l and purified to homogeneity using Hisbinding nickel-agarose affinity chromatography. The purified enzyme was maximally active at $70^{\circ}C$, pH 4.5, and lost no activity over 10 days at $50^{\circ}C$. ${\alpha}$-Gal1 followed Michaelis-Menten kinetics ($V_{max}\;of\;240.3{\mu}M/min/mg,\;K_m\;of\;0.294 mM$) and was inhibited competitively by galactose ($K_m{^{obs}}$ of 0.57 mM, $K_i$ of 2.77 mM). The recombinant T. emersonii ${\alpha}$-galactosidase displayed broad substrate preference, being active on both oligo- and polymeric substrates, yet had strict specificity for the ${\alpha}$-galactosidic linkage. Owing to its substrate preference and noteworthy stability, ${\alpha}$-Gal1 is of particular interest for possible biotechnological applications involving the processing of plant materials.

Characterization of αX I-Domain Binding to Receptors for Advanced Glycation End Products (RAGE)

  • Buyannemekh, Dolgorsuren;Nham, Sang-Uk
    • Molecules and Cells
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    • v.40 no.5
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    • pp.355-362
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    • 2017
  • The ${\beta}2$ integrins are cell surface transmembrane proteins regulating leukocyte functions, such as adhesion and migration. Two members of ${\beta}2$ integrin, ${\alpha}M{\beta}2$ and ${\alpha}X{\beta}2$, share the leukocyte distribution profile and integrin ${\alpha}X{\beta}2$ is involved in antigen presentation in dendritic cells and transendothelial migration of monocytes and macrophages to atherosclerotic lesions. ${\underline{R}}eceptor$ for ${\underline{a}}dvanced$ ${\underline{g}}lycation$ ${\underline{e}}nd$ ${\underline{p}}roducts$ (RAGE), a member of cell adhesion molecules, plays an important role in chronic inflammation and atherosclerosis. Although RAGE and ${\alpha}X{\beta}2$ play an important role in inflammatory response and the pathogenesis of atherosclerosis, the nature of their interaction and structure involved in the binding remain poorly defined. In this study, using I-domain as a ligand binding motif of ${\alpha}X{\beta}2$, we characterize the binding nature and the interacting moieties of ${\alpha}X$ I-domain and RAGE. Their binding requires divalent cations ($Mg^{2+}$ and $Mn^{2+}$) and shows an affinity on the sub-micro molar level: the dissociation constant of ${\alpha}X$ I-domains binding to RAGE being $0.49{\mu}M$. Furthermore, the ${\alpha}X$ I-domains recognize the V-domain, but not the C1 and C2-domains of RAGE. The acidic amino acid substitutions on the ligand binding site of ${\alpha}X$ I-domain significantly reduce the I-domain binding activity to soluble RAGE and the alanine substitutions of basic amino acids on the flat surface of the V-domain prevent the V-domain binding to ${\alpha}X$ I-domain. In conclusion, the main mechanism of ${\alpha}X$ I-domain binding to RAGE is a charge interaction, in which the acidic moieties of ${\alpha}X$ I-domains, including E244, and D249, recognize the basic residues on the RAGE V-domain encompassing K39, K43, K44, R104, and K107.

Nutritional Value and Bioactive Properties of Enzymatic Hydrolysates prepared from the Livers of Oncorhynchus keta and Oncorhynchus gorbuscha (Pacific Salmon)

  • Yoon, Ho Dong;Karaulova, Ekaterina P.;Shulgina, Lilia V.;Yakush, Evgeni V.;Mok, Jong Soo;Lee, Su Seon;Xie, Chengliang;Kim, Jeong Gyun
    • Fisheries and Aquatic Sciences
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    • v.18 no.1
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    • pp.13-20
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    • 2015
  • Calculated chemical scores (computed in relation to the FAO/WHO reference protein) for salmon liver protein hydrolysates indicated that all amino acids (other than methionine and threonine) were present in adequate or excess quantities; thus, the raw liver material is a good source of essential amino acids. The hydrophobic amino acids contents in hydrolysates prepared from Oncorhynchus keta and O. gorbuscha were 38.4 and 39.1%, respectively. The proportion of released peptides exceeding 500 kDa was reduced when hydrolysates were treated with the commercial enzyme Alcalase, although proportions in the following MW ranges were elevated: 100-500 kDa and <50 kDa. The optimal conditions for enzymatic hydrolysis were as follows: pH 7.0, $50^{\circ}C$, and a reaction time of 1 h. Of the different proteases tested, Alcalase was the most efficient for production of salmon liver hydrolysate with the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity. The hydrolysates prepared from salmon liver had a balanced amino acid composition. The liver protein hydrolysates contained low molecular weight peptides, some of which may be bio-active; this bio-active potential should be investigated. Inhibition of the DPPH radical increased with increased degree of hydrolysis (DH), regardless of protease type. DPPH radical scavenging abilities, antithrombotic effects and ${\alpha}$-glucosidase enzyme inhibition effects of O. keta liver hydrolysate increased in a dose-dependent manner. Thus, salmon liver hydrolysate may be useful in functional food applications and as a source of novel products.

AtMAP65-1 Binds to Tubulin Dimers to Promote Tubulin Assembly

  • Li, Hua;Yuan, Ming;Mao, Tonglin
    • BMB Reports
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    • v.40 no.2
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    • pp.218-225
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    • 2007
  • In Arabidopsis thaliana, the microtubule-associated protein AtMAP65-1 shows various functions on microtubule dynamics and organizations. However, it is still an open question about whether AtMAP65-1 binds to tubulin dimers and how it regulates microtubule dynamics. In present study, the tubulin-binding activity of AtMAP65-1 was investigated. Pull-down and co-sedimentation exp eriments demonstrated that AtMAP65-1 bound to tubulin dimers,at a molar ratio of 1 : 1. Cross-linking experiments showed that AtMAP65-1 bound to tubulin dimers by interacting with $\alpha$-tubulin of the tubulin heterodimer. Interfering the bundling effect of AtMAP65-1 by addition of salt and monitoring the tubulin assembly, the experiment results indicated that AtMAP65-1 promoted tubulin assembly by interacting with tubulin dimers. In addition, five truncated versions of AtMAP65-1, namely AtMAP65-1 $\Delta$N339 (amino acids 340-587); AtMAP65-1 $\Delta$N494 (amino acids 495-587); AtMAP65-1 340-494 (amino acids 340-494); AtMAP65-1 $\Delta$C495 (amino acids 1-494) and AtMAP65-1 $\Delta$C340 (amino acids 1-339), were tested for their binding activities and roles in tubulin polymerization in vitro. Four (AtMAP65-1 $\Delta$N339, $\Delta$N494, AtMAP65-1 340-494 and $\Delta$C495) from the five truncated proteins were able to co-sediment with microtubules, and three (AtMAP65-1 $\Delta$N339, $\Delta$N494 and AtMAP65-1 340-494) of them could bind to tubulin dimers in vitro. Among the three truncated proteins, AtMAP65-1 $\Delta$N339 showed the greatest activity to promote tubulin polymerization, AtMAP65-1 $\Delta$N494 exhibited almost the same activity as the full length protein in promoting tubulin assembly, and AtMAP65-1 340-494 had minor activity to promote tubulin assembly. On the contrast, AtMAP65-1 $\Delta$C495, which bound to microtubules but not to tubulin dimers, did not affect tubulin assembly. Our study suggested that AtMAP65-1 might promote tubulin assembly by binding to tubulin dimers in vivo.

Current Studies on Browning Reaction Products and Acidic Polysaccharide in Korean Red Ginseng (홍삼에 함유된 갈변물질 및 산성다당체에 대한 연구현황)

  • Lee, Jong-Won;Do, Jae-Ho
    • Journal of Ginseng Research
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    • v.30 no.1
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    • pp.41-48
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    • 2006
  • In the browning reaction of Korean ginseng, it appears that enzymatic and non-enzymatic browning reaction occurred In initial stage of steaming fresh ginseng at low temperature, and then non-enzymatic browning reaction followed in the drying period after steaming. Browning reaction of red ginseng occurred between $60{\sim}90$ min of steaming at $100^{\circ}C$, and browning pigments of red ginseng were mostly water soluble substances. The structural characteristics of water soluble browning reaction products(WS-BRPs) isolated from Korean red ginseng were showed the presence of hydroxyl, amide carbonyl and aliphatic methane groups. From sugar analysis it was identified that L and S-1, melanoidins isolated from red ginseng, contained two kinds of sugars, glucose and xylose, and the other melanoidin S-2 contained the previous and fructose. In order to find out pertinent methods for the acceleration of browning during ginseng processing, various treatment were made on fresh ginseng with sugars, amino acids and inorganic nitrogenous compounds and the extent of browning was measured. Among sugar tested, maltose resulted in the greatest acceleration of browning followed in decreasing order by glucose and lactose, whereas pentoses, fructose, sucrose and raffinose had negligible effect. A marked browning occurred in ginseng treated with basic amino acids, while the extent of browning was not greatly increased when ginseng was treated with aliphatic amino acids, hydroxyl amino acids, or acidic amino acids. The brown color intensity gradually increased with an increase of glucose concentration far up to 0.5M. L, S-1, and S-2 were found to have an ability to donate hydrogen to DPPH, and also they had anti-oxidative activity in the experiments of hydrogen peroxide scavenging, inhibitory activity in the formation of MDA from linoleic acid, auto oxidation of ok-brain homogenates, lipid peroxidation by the enzymatic and non-enzymatic system in liver microsome fraction, and mitochondrial fraction etc. The amounts of acidic polysaccharide(AP) in red ginseng were higher than those of wild and cultured Panax quinquefolius, Panax notoginseng as well as white ginseng (Panax ginseng). In white ginseng, the AP amount is no difference in root ages or sizes, also, the AP amount of ginseng body was similar to that of rhizome, but was higher than that of leaf and epidermis. Addition of red ginseng acidic polysaccharide(RGAP) increased production of nitric oxide(NO) and tumor necrosis factor (TNF)-$\alpha$ in the rodent macrophage cultures, and treatment of RGAP in vivo stimulated tumoricidal activities of natural killer (NK) cells.

Investigations on Possible Roles of C-Terminal Propeptide of a Ca-Independent ${\alpha}$-Amylase from Bacillus

  • Salimi, Ali;Yousefi, Fatemeh;Ghollasi, Marzieh;Daneshjou, Sara;Tavoli, Hesam;Ghobadi, Sirous;Khajeh, Khosro
    • Journal of Microbiology and Biotechnology
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    • v.22 no.8
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    • pp.1077-1083
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    • 2012
  • Previously, an extracellular ${\alpha}$-amylase (BKA) had been purified from the culture of Bacillus sp. KR8104. Subsequently, the crystal structure of the active enzyme revealed a 422 amino acids polypeptide. In this study, the bka was cloned into E. coli, which encoded a polypeptide of 659 amino acids including two additional fragments: one 44 residues N-terminal fragment and another 193 residues C-terminal fragment. In order to investigate the role of the C-terminal fragment, two constructs with and without this region [$BKA{\Delta}$(N44) and $BKA{\Delta}$(N44C193)] were designed and expressed in E. coli BL21. The optimum pH, thermal stability, and the end-products of starch hydrolysis were found to be similar in both constructs. The $K_m$ and $V_{max}$ values for $BKA{\Delta}$(N44) were lower than $BKA{\Delta}$(N44C193), using either starch or ethylidene-blocked 4-nitrophenylmaltoheptaoside as a substrate.