• Title/Summary/Keyword: $\beta$-Amino acid

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Isolation, Purification and Characterization of the β-Xylosidase from Klebsiella sp. Sc. (Klebsiella sp. Sc가 생산하는 β-xylosidase의 분리, 정제 및 특성)

  • Lee, Yong-Seok;Park, In-Hye;Ahn, Soon-Cheol;Choi, Yong-Lark
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1801-1806
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    • 2010
  • A $\beta$-xylosidase encoding gene from Klebsiella sp. Sc was cloned in Escherichia coli. The $\beta$-xylosidase gene consisted of an open reading frame of 1,680 nucleotides and encodes 559 amino acids with a deduced molecular weight of 63 kDa. The deduced amino acid sequence of the $\beta$-xylosidase from Klebsiella sp. Sc exhibits 90% identities and 95% positives compared to those from Klebsiella oxytoca (KOX), Lactobacillus lactis (LAC, 82%, 90%), Bacillus longum (BLON, 69%, 81%) and Escherichia coli (ECOLI, 47%, 63%). The $\beta$-xylosidase was purified by GST-fusion purification system. The pH and temperature optima of the enzyme were 6.6 and $55^{\circ}C$, respectively. The $\beta$-xylosidase hydrolyzes xylobiose to xylose.

Bacillus cereus ASK-202에서 cloning 된 agarase의 물리 ${\cdot}$ 화학적 특성

  • Hwang, Seon-Hui;Ha, Sun-Deuk;Kim, Bong-Jo;Gong, Jae-Yeol
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.534-537
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    • 2002
  • An agarase gene from Bacillus cereus ASK202 was expressed in high levels by E. coli BL21(DE3)/pEBA1 using pET28a(+) vector system with the inducible T7 promoter in the presence of isopropyl- ${\beta}$ -thiogalactopyranoside. The open reading frame encodes 761 amino acid residues with a calculated molecular weight of 83,300 daltons and a potential signal peptide about 36 amino acid residues at the N-terminus. E. coli BL21(DE3)/pEBA1 produce 1280 unit/ ${\ell}$ of agarase. The optimum physical condition for the agarase activity was pH 5.6, and $40^{\circ}C$, respectively. The agarase activity was stable up pH $4.0{\sim}9.0$ and $4{\sim}40^{\circ}C$. The km and maximum rate of metabolism for agar were 0.068mg/$m{\ell}$ and 0.094mg/$m{\ell}{\cdot}min$, respectively.

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Establishment of an In Vivo Report System for the Evaluation of Amber Suppression Activity in Escherichia coli (대장균에서 비천연 아미노산의 위치특이적 삽입을 위한 Amber Suppressor tRNA와 Aminoacyl-tRNA Synthetase의 Amber Suppression 활성측정시스템 개발)

  • Kim, Kyung-Tae;Park, Mi-Young;Park, Jung-Chan
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.215-221
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    • 2009
  • Site-specific incorporation of unnatural amino acids into proteins in vivo can be achieved by co-expression of an orthogonal pair of suppressor tRNA and engineered aminoacyl-tRNA synthetase (ARS) that specifically ligates an unnatural amino acid to the suppressor tRNA. As a step to establish this technique, here we generated an Escherichia coli reporter strain DH10B(Tn:lacZam) by integrating amber mutated lacZ gene into the chromosome of E. coli DH10B strain. In vivo expression of E. coli amber suppressor $tRNA^{Gln}$ produced blue colonies in culture plates containing X-Gal as well as dramatically increased $\beta$-galactosidase activity. In addition, expression of an orthogonal pair of Saccharomyces cerevisiae suppressor $tRNA^{Tyr}$ and tyrosyl-tRNA synthetase also produced blue colonies as well as moderate increase of $\beta$-galactosidase activity. These data demonstrate that our reporter strain will provide an efficient method to assess amber suppression in both qualitative and quantitative manners.

Cloning, Expression, and Characterization of a Novel GH-16 β-Agarase from Agarivorans sp. JA-1 (Agarivorans sp. JA-1 유래 신규 GH-16 β-agarase의 클로닝, 발현 및 특성)

  • Jeon, Myong Je;Kim, A-Ram;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1545-1551
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    • 2012
  • Authors report the glycoside hydrolase (GH) family 16 ${\beta}$-agarase from the strain of Agarivorans sp. JA-1, which authors previously stated as recombinant expression and characterization of GH-50 and GH-118 ${\beta}$-agarase. It comprised an open reading frame of 1,362 base pairs, which encodes a protein of 49,830 daltons consisting of 453 amino acid residues. Valuation of the total sequence showed that the enzyme has 98% nucleotide and 99% amino acid sequence similarities to those of GH-16 ${\beta}$-agarase from Pseudoalteromonas sp. CY24. The gene corresponding to a mature protein of 429 amino acids was recombinantly expressed in Escherichia coli, and the enzyme was purified to homogeneity by affinity chromatography. It showed maximal activity at $40^{\circ}C$ and pH 5.0, representing 67.6 units/mg. Thin layer chromatography revealed that mainly neoagarohexaose and neoagarotetraose were produced from agarose. The enzyme would be valuable for the industrial production of functional neoagarooligosaccharides.

A Novel All-trans Retinoid Acid Derivative Induces Apoptosis in MDA-MB-231 Breast Cancer Cells

  • Wang, Bei;Yan, Yun-Wen;Zhou, Qing;Gui, Shu-Yu;Chen, Fei-Hu;Wang, Yuan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10819-10824
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    • 2015
  • Aims: To explore the effect and probable mechanism of a synthetic retinoid 4-amino-2-tri-fluoromethylphenyl ester (ATPR) on apoptosis of MDA-MB-231 breast cancer cells. Materials and Methods: MTT assays were performed to measure the proliferation of MDA-MB-231 cells treated with different concentrations of all-trans retinoic acid (ATRA) and ATPR. Morphologic changes were observed by microscopy. The apoptosis rates and cell cycling of MDA-MB-231 cells treated with ATRA or ATPR were assessed using flow cytometry analysis. Expression of retinoic acid receptor and phosphorylation of ERK, JNK, p38 proteins were detected by Western blotting. Results: Treatment of the cells with the addition of $15{\mu}mol/L$ ATPR for 48 h clearly demonstrated reduced cell numbers and deformed cells, whereas no changes in the number and morphology were observed after treatment with ATRA. The apoptosis rate was 33.2% after breast cancer MDA-MB-231 cells were treated by ATPR ($15{\mu}mol/L$) whereas ATRA ($15{\mu}mol/L$) had no apoptotic effect. ATPR inhibited the phosphorylation of ERK, JNK, and p38 while ATRA had no significant effect. ATPR inhibited the expression of BiP and increased the expression of Chop at the protein level compared with control groups, ATRA and ATPR both decreased the protein expression of $RXR{\alpha}$, ATPR reduced the protein expression of $RAR{\beta}$ and $RXR{\beta}$ while ATRA did not decrease $RAR{\beta}$ or $RXR{\beta}$. Conclusions: ATPR could induce apoptosis of breast cancer MDA-MB-231 cells, possible mechanisms being binding to $RAR{\beta}/RXR{\beta}$ heterodimers, then activation of ER stress involving the MAPK pathway.

Comparison of The Taste Compounds of Wild and Cultured Eel, Puffer and Snake head (천연산 및 양식산 뱀장어, 복어, 가물치의 맛 성분에 대한 연구)

  • Kim, Hee-Yun;Shin, Jae-Wook;Sim, Gyu-Chang;Park, Hee-Ok;Kim, Hyun-Sook;Kim, Sang-Moo;Cho, Jae-Sun;Jang, Young-Mi
    • Korean Journal of Food Science and Technology
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    • v.32 no.5
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    • pp.1058-1067
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    • 2000
  • In this study, eel, puffer and snake head fish, which are widely taken and highly valued for processing into baked fish, soup or fish juice were studied by classifying them into wild fish and cultured fish, and evaluated in terms of taste. The samples were gathered in the area including Kimhae, Samchonpo and Haman in Kyungsangnam-do, Kangsu-Gu in Pusan and Bukcheju-Gun in Cheju-do. Proximate compositions, the content of nucleotides and their related compounds, total and free amino acids, organic acids and fatty acids of the fishes were analysed. Generally, as for the compositions, wild fish had a higher crude fat content and a lower moisture content as compared with cultured fish, while there was no great difference between them in terms of crude protein and ash contents. Nucleotides and their related compounds including ATP, ADP, AMP, IMP, HxR, Hx were detected, and the amounts of which were nearly the same with respect to the growth conditions. IMP content was high in all of the samples, while ATP content was extremely low. Total 17 amino acids were detected from the samples, and most of the samples had high contents of glutamic acid, aspartic acid, lysine, leucine, glycine, alanine and arginine, and low contents of cystein, isoleucine and methionine. As for the other amino acids, generally same level of contents were detected for each samples. In cases of eel and snake head fish, the wild fish had higher total amino acid contents in comparison with cultured fish. And the result to the contrary was obtained for the case of puffer. Sample fishes had nearly the same compose distribution of free amino acid though the contents of which were a little different according to the samples. Each sample had high contents of taurine, hydroxyproline, glutamic acid, glycine, cystathionine, ${\beta}-aminoisobutyric$ acid and lysine. Total organic acid contents of cultured fish was higher than that of wild fish. All fishes commonly had high contents of lactic acid, iso-valeric acid, oxalic acid and fumaric acid. High levels of $C_{16:0},\;C_{16:1},\;C_{18:1(cis)},\;C_{18:3},\;and\;C_{22:6}$ were shown in all of the samples and ${\omega}-3$ polyunsaturated fatty acid content of cultured fish was higher than that of wild fish, and as for the essential fatty acid, wild fish has higher content in comparison with cultured fish.

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Association Between $\beta_2$ Adrenoceptor Polymorphisms and Atopy/serum IgE in Asthmatic Patients ($\beta_2$ 교감신경 수용체 다형성이 아토피 및 혈청 IgE에 미치는 영향)

  • Lee, Sin-Hyung;Shim, Jae-Jeong;Kang, Yong-Koo;Jeong, Hye-Cheol;Kim, Kyung-Kyu;Kwon, Young-Hwan;Kim, Je-Hyeong;Lee, Sung-Yong;Lee, So-Ra;Lee, Sang-Youb;Cho, Jae-Youn;In, Kwang-Ho;Yoo, Sa-Hwa;Kang, Kyung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.6
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    • pp.826-835
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    • 1999
  • Background : The $\beta_2$ adrenergic receptor ($\beta_2$ AR) polymorphisms occurring at amino acid position 16 (Arg to Gly), 27 (Gln to Glu), 34 (Val to Met), and 164 (Thr to Ile) are known to be functionally relevant and also disease-modifying in subjects with asthma. However the contribution of these polymorphisms to the development of the asthmatic phenotype or other markers for allergic disease remains to be established. Methods : 109 patients with bronchial asthma and 42 healthy person were included. Serum total IgE, allergen specific IgE, and skin prick test were performed to all of the subjects. $\beta_2$ AR polymorphisms were checked by mutated allele specific amplification (MASA) method. Results : The results were as follows. The frequencies of $\beta_2$ AR polymorphisms in asthmatic patients and healthy person were not statistically different(p>0.05). There was no association between $\beta_2$ AR polymorphisms of amino acid position 16, 27, 34 and the existence of atopy among asthmatic patients(p>0.05). Between asthmatic patients with or without elevated IgE level and $\beta_2$ AR polymorphisms of amino acid position 16, 27, 34, there was no statistically significant association(p>0.05). Conclusion : There was no difference in frequency of the $\beta_2$ AR polymorphism between asthmatic patients and healthy person. In the bronchial asthma, association of $\beta_2$ AR polymorphism and atopy/serum total IgE was not found.

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Effect of Single Amino Acid Replacements on the Folding of $\alpha_1$-Antitrypsin

  • Lee, Cheolju;Yu, Myeong-Hee
    • Proceedings of the Korean Biophysical Society Conference
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    • 1998.06a
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    • pp.24-24
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    • 1998
  • The effect of stabilizing single ammo acid replacements at the sites of Phe 51, Ala 70, and Met 374 on the folding of $\alpha$$_1$-antitrypsin was investigated by fluorescence spectroscopy. The residues Phe 51 and Met 374 are located in the hydrophobic core of the molecule, B $\beta$-sheet.(omitted)

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Flavor development in cheddar cheese (체다 치즈의 맛의 개발)

  • 정청송;유상훈
    • Journal of Applied Tourism Food and Beverage Management and Research
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    • v.14 no.1
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    • pp.59-77
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    • 2003
  • This study was carried out to find a cholesterol removal rate, flavor development, and bitter amino acid productions in Cheddar cheese treated with -cyclodextrin ($\beta$-CD): l) Control (no homogenization, no $\beta$-CD), and 2) Milk treatment (1000 psi milk homogenization, 1 % $\beta$-CD). The cholesterol removal of the cheese were 79.3%. The production of short-chain free fatty acids (FF A) increased with a ripening time in both control and milk treated cheese. The releasing quantity of short-chain FFA was higher din milk treated cheese than control at 5 and 7 mo ripening. Not much difference was found in neutral volatile compounds production between samples. In bitter-tasted amino acids, milk treatment group produced much higher than control. In sensory analysis, texture score of control Cheddar cheese significantly increased, however, that in cholesterol-reduced cheese decreased dramatically with ripening time.

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Bombyx mori β-tubulin Promoter for High-level Expression of Heterologous Genes

  • Park, Kwanho;Goo, Tae-Won
    • International Journal of Industrial Entomology and Biomaterials
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    • v.39 no.1
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    • pp.22-28
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    • 2019
  • We previously isolated 9 clones that show stronger signal compared to Bombyx mori cytoplasmic actin gene (BmA3) by using a dot blot hybridization. In this study, we focused on one clone among these clones which has high amino acid similarity with ${\beta}$-tubulin gene of B. mori. This clone was ubiquitously expressed in all tissues and developmental stage of B. mori. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-750/-1) in the 5'-flanking region of ${\beta}$-tubulin gene, which has about 47 fold more intensive promoter activity than BmA3 promoter. Moreover, the ${\beta}$-tubulin promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that ${\beta}$-tubulin promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter.