• Title/Summary/Keyword: Functional Amino Acids

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Nucleotide Sequence of Rat Transketolase and Liver-Specific Pretranslational Activation During Postnatal Development

  • Kim, Sung-Min F.;Kim, Byung-Moon;Jeng, Jingjau;Soh, Yun-Jo;Bak, Choong-Il;Huh, Jae-Wook;Song, Byoung-J.
    • BMB Reports
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    • v.29 no.2
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    • pp.146-150
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    • 1996
  • A 2.1 kb cDNA clone for rat transketolase was isolated from rat liver ${\lambda}gt11$ cDNA library and its sequence was determined. The predicted rat transketolase (655 amino acids with $M_r$ 71,186) is highly similar (92%) to that of the human enzyme except that it contains an extra 32 amino acids at its N-terminus. Although it is less similar (<27%) to transketolases from non-mammalian species, the functional motifs such as the catalytic sites and thiamine binding domain are well conserved in the rat enzyme. Southern blot analysis of genomic DNA verified that transketolase appears to be derived from a single gene. Immunoblot and Northern blot analyses suggested that hepatic transketolase was activated pretranslationally by a 2.1-fold while little change was observed in brain enzyme, indicating a tissue-specific pretranslational activation during postnatal development.

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Identification and Heterologous Expression of a ${\Delta}4$-Fatty Acid Desaturase Gene from Isochrysis sphaerica

  • Guo, Bing;Jiang, Mulan;Wan, Xia;Gong, Yangmin;Liang, Zhuo;Hu, Chuanjiong
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1413-1421
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    • 2013
  • The marine microalga Isochrysis sphaerica is rich in the very-long-chain polyunsaturated fatty acids, including eicosapentaenoic acid (EPA, $C20:5{\omega}-3$) and docosahexaenoic acid (DHA, $C22:6{\omega}-3$) that are important to human health. Here, we report a functional characterization of a ${\Delta}4$-fatty acid desaturase gene (FAD4) from I. sphaerica. IsFAD4 contains a 1,284 bp open reading frame encoding a 427 amino acid polypeptide. The deduced amino sequence comprises three conserved histidine motifs and a cytochrome b5 domain at its N-terminus. Phylogenetic analysis indicated that IsFad4 formed a unique Isochrysis clade distinct from the counterparts of other eukaryotes. Heterologous expression of IsFAD4 in Pichia pastoris showed that IsFad4 was able to desaturate docosapentaenoic acid (DPA) to form DHA, and the rate of converting DPA to DHA was 79.8%. These results throw light on the potential industrial production of specific polyunsaturated fatty acids through IsFAD4 transgenic yeast or oil crops.

Whitening and Anti-aging Activities of Soluble Silkworm Gland Hydrolysate (수용성 누에 실샘 가수분해물의 미백 및 항노화 효과)

  • Jung, Ji-Hye;Hwang, Jung Wook;Kim, Hojin;Cha, Hyun-Myoung;Kim, Dong-Il;Choi, Yong-Soo
    • KSBB Journal
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    • v.28 no.3
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    • pp.196-201
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    • 2013
  • Silkworm-derived silk materials which contain 45 to 55% protein (18 amino acids, including 8 essential amino acids) have free radical-scavenging activities. In the present study, we investigated the whitening and anti-aging effects of silkworm gland under various conditions with B16F1 melanoma cells and human dermal fibroblasts. To evaluate the toxicity of soluble gland hydrolysate (SGH), mouse-derived B16F1 melanoma cells were treated with 0.2~5 mg/mL, cytotoxicity was not observed at all concentrations. The tyrosinase and elastase activities were significantly decreased as dosage levels of SGH increased. In addition, cell death was decreased by treated with SGH and antioxidative activity was the most effective in the SGH treatment. These findings suggest that SGH may be used as a potential functional biomaterial in having the skin whitening effect and anti-aging activity by inhibition of tyrosinase and elastase activities.

The antioxidant roles of L-carnitine and N-acetyl cysteine against oxidative stress on human sperm functional parameters during vitrification

  • Ghorbani, Fatemeh;Nasiri, Zohreh;Koohestanidehaghi, Yeganeh;Lorian, Keivan
    • Clinical and Experimental Reproductive Medicine
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    • v.48 no.4
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    • pp.316-321
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    • 2021
  • Objective: Amino acids can protect sperm structure in cryopreservation due to their antioxidant properties. Therefore, the present study aimed to investigate the protective effect of L-carnitine (LC) and N-acetyl cysteine (NAC) on motility parameters, plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), DNA damage, and human sperm intracellular reactive oxygen species (ROS) during vitrification. Methods: Twenty normal human sperm samples were examined. Each sample was divided into six equal groups: LC (1 and 10 mM), NAC (5 and 10 mM), and cryopreserved and fresh control groups. Results: The groups treated with LC and NAC showed favorable findings in terms of motility parameters, DNA damage, and MMP. Significantly higher levels of intracellular ROS were observed in all cryopreserved groups than in the fresh group (p≤0.05). The presence of LC and NAC at both concentrations caused an increase in PMI, MMP, and progressive motility parameters, as well as a significant reduction in intracellular ROS compared to the control group (p≤0.05). The concentrations of the amino acids did not show any significant effect. Conclusion: LC and NAC are promising as potential additives in sperm cryopreservation.

Quality Characteristics of Bread with Outdoor Cultivated Bigeumdo Seomcho (Spinacia oleracea L.) Powder (노지재배 비금도 섬초(시금치) 분말을 첨가한 식빵의 품질 특성)

  • Kim, Young-Mo;Yun, Yong-Chul
    • The Korean Journal of Food And Nutrition
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    • v.34 no.4
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    • pp.321-330
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    • 2021
  • The purpose of this study was to investigate the quality characteristics of bread made with 1%, 3%, 5%, and 7% of Bigeumdo Seomcho powder (Spinacia oleracea L.). The pH of dough and bread decreased as the amount of addition increased; 7% indicated the lowest. The moisture level correlated negatively with the powder addition, but the difference was not statistically meaningful (p<0.05). As the amount of addition was increased, the intensity of color increased but the volume decreased. The brightness and redness of the bread decreased as the amount of addition increased. The volume, specific volume, and baking loss rate correlated negatively with the powder addition. Regarding springiness, cohesiveness, and gumminess, the experiment group was higher than the control group and they lowered as the addition amount increased. The brittleness showed positive correlation with the addition amount. Out of the amino acids of bread, glutamic acid showed the highest value as the amount of addition was increased. Overall, it is possible to produce bakery bread using functional ingredients.

Bovine Lactoferricin Induces Intestinal Epithelial Cell Activation through Phosphorylation of FAK and Paxillin and Prevents Rotavirus Infection

  • Jeong, Ye Young;Lee, Ga Young;Yoo, Yung Choon
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1175-1182
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    • 2021
  • We investigated the effect of bovine lactoferricin (Lfcin-B), a peptide derived from bovine lactoferrin, on activation of intestinal epithelial cells in IEC-6 intestinal cell, and protection against in vivo rotavirus (RV) infection. Treatment with Lfcin-B significantly enhanced the growth of IEC-6 cells and increased their capacity for attachment and spreading in culture plates. Also, Lfcin-B synergistically augmented the binding of IEC-6 cells to laminin, a component of the extracellular matrix (ECM). In the analysis of the intracellular mechanism related to Lfcin-B-induced activation of IEC-6 cells, this peptide upregulated tyrosine-dependent phosphorylation of focal adhesion kinase (FAK) and paxillin, which are intracellular proteins associated with cell adhesion, spreading, and signal transduction during cell activation. An experiment using synthetic peptides with various sequences of amino acids revealed that a sequence of 9 amino acids (FKCRRWQWR) corresponding to 17-25 of the N-terminus of Lfcin-B is responsible for the epithelial cell activation. In an in vivo experiment, treatment with Lfcin-B one day before RV infection effectively prevented RV-induced diarrhea and significantly reduced RV titers in the bowels of infected mice. These results suggest that Lfcin-B plays meaningful roles in the maintenance and repair of intestinal mucosal tissues, as well as in protecting against intestinal infection by RV. Collectively, Lfcin-B is a promising candidate with potential applications in drugs or functional foods beneficial for intestinal health and mucosal immunity.

Characterization of dihydroflavonol 4-reductase cDNA in tea [Camellia sinensis (L.) O. Kuntze]

  • Singh, Kashmir;Kumar, Sanjay;Yadav, Sudesh Kumar;Ahuja, Paramvir Singh
    • Plant Biotechnology Reports
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    • v.3 no.1
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    • pp.95-101
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    • 2009
  • Tea leaves are major source of catechins—antioxidant flavonoids. Dihydroflavonol 4-reductase (DFR, EC 1.1.1.219) is one of the important enzymes that catalyzes the reduction of dihydroflavonols to leucoanthocyanins, a key ''late'' step in the biosynthesis of catechins. This manuscript reports characterization of DFR from tea (CsDFR) that comprised 1,413 bp full-length cDNA with ORF of 1,044 bp (115-1,158) and encoding a protein of 347 amino acids. Sequence comparison of CsDFR with earlier reported DFR sequences in a database indicated conservation of 69-87% among amino acid residues. In silico analysis revealed CsDFR to be a membrane-localized protein with a domain (between 16 and 218 amino acids) resembling the NAD-dependent epimerase/dehydratase family. The theoretical molecular weight and isoelectric point of the deduced amino sequence of CsDFR were 38.67 kDa and 6.22, respectively. Upon expression of CsDFR in E. coli, recombinant protein was found to be functional and showed specific activity of 42.85 nmol $min^{-1}$ mg $protein^{-1}$. Expression of CsDFR was maximum in younger rather than older leaves. Expression was down-regulated in response to drought stress and abscisic acid, unaffected by gibberellic acid treatment, but up-regulated in response to wounding, with concomitant modulation of catechins content. This is the first report of functionality of recombinant CsDFR and its expression in tea.

Amino acid substitution on β and α of Cyt2Aa2 affects molecular interaction of protoxin

  • Thammachat, Siriya;Pungtanom, Nuanwan;Kidsanguan, Somruathai;Pathaichindachote, Wanwarang;Promdonkoy, Boonhiang;Krittanai, Chartchai
    • BMB Reports
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    • v.43 no.6
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    • pp.427-431
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    • 2010
  • Cyt2Aa2 is a mosquito-larvicidal protein produced as a 29 kDa crystalline protoxin from Bacillus thuringiensis subsp. darmstadiensis. To become an active toxin, proteolytic processing is required to remove amino acids from its N- and C-termini. This study aims to investigate the functional role of amino acid residues on the N-terminal ${\beta}1$ and C-terminal ${\alpha}F$ of Cyt2Aa2 protoxin. Mutant protoxins were constructed, characterized and compared to the wild type Cyt2Aa2. Protein expression data and SDS-PAGE analysis revealed that substitution at leucine-33 (L33) of ${\beta}1$ has a critical effect on dimer formation and structural stability against proteases. In addition, amino acids N230 and I233-F237 around the C-terminus ${\alpha}F$ demonstrated a crucial role in protecting the protoxin from proteolytic digestion. These results suggested that ${\beta}1$ and ${\alpha}F$ on the Nand C-terminal ends of Cyt2Aa2 protoxin play an important role in the molecular interaction and in maintaining the structural stability of the protoxin.

Studies on Physiological and Functional Properties of Susijang, Fermented Soybean Paste (수시장(콩발효식품)의 생리기능적 특성에 관한 연구)

  • Ryu, Bog-Mi;Sugiyama, Kuniko;Kim, Jung-Suk;Park, Min-Hee;Moon, Gap-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.2
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    • pp.137-142
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    • 2007
  • This study was carried out to observe the antioxidant and angiotensin converting enzyme (ACE) inhibitory activity of susijang (Korean traditional fermented soy food). The antioxidant activity of susijang was measured by using TEAC (trolox equivalent antioxidant capacity) assay. In addition contents of isoflavones, phenolic acids and amino acids were determined. TEAC values of chungkukjang and susijang were significantly higher than those of yellow soybean (p<0.05). The contents of isoflavones (genistein, daidzein) were found to be significantly higher in chungkukjang and susijang than yellow soybean. The susijang showed high content of free amino acids and phenolic acids. ACE inhibitory activity of susijang was significantly higher than those of chungkukjang and yellow soybean, particularly of 70% ethanol extract.

Compositional Characteristics of the Microalga Melosira nummuloides Mass-cultured Using Jeju Lava Seawater (제주도 용암 해수로 대량배양한 미세조류 멜로시라(Melosira nummuloides)의 성분 특성)

  • Kim, Yunyoung;Shin, Hyun-A;Choi, Jeong-Woo;Kim, Mi-Yeon;Go, Gyung-Min
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.55 no.2
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    • pp.91-101
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    • 2022
  • In this study, we analyzed the compositional characteristics of Melosira nummuloides mass-cultured using Jeju lava seawater. M. nummuloides showed the highest growth rate when cultured for 14 days at 17-20℃ and 15,000 lx. Proximate composition of raw-material (RM) and freeze-dried M. nummuloides (FM) showed high ash content (65-72%), while ethanol-extracted M. nummuloides (EM) had low ash and high lipid contents. The predominantly occurring mineral, Si, was 334 g/kg in RM and 269 g/kg in FM, but EM contained only 1.97 g/kg. The ratio of essential amino acids was similar in RM (38.93%) and FM (36.89%) lower in EM (17.83%), but branched chain amino acids required for muscle metabolism was high (63.40%). The polyunsaturated fatty acids of EM (34.74%) was 11% more than that in RM (23.81%), and the ratio of omega-6 to omega-3 fatty acids was 1:3.6. Furthermore, the concentration of total chlorophyll was higher by 5.15 times (62.32 mg/g) and that of fucoxanthin by 7.06 times (11.02 mg/g) in EM than in RM (12.11, 1.56 mg/g, respectively). The mass cultivation and filtration of M. nummuloides using lava seawater has high prospects of commercialization as a competitive bio silica, cosmetic and healthy functional food material.