• Title/Summary/Keyword: Biological Synthesis

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Synthesis and Biological Properties of l$\beta$-Methylcarbapenems with 5'- Ethoxycarbonylmethoxy(hydroxy )iminopyrrolidinylthio Group at C-2 Position (C-2 위치에 5'-Ethoxycarbonylmethoxy(hydroxy)iminopyrrolidinylthio Group을 가진 1$\beta$-Methylcarbapenems의 합성과 생물학적 성질)

  • 오창현;안수현;백대진;홍준희;고옥현
    • YAKHAK HOEJI
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    • v.47 no.6
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    • pp.450-455
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    • 2003
  • Synthesis of (lR,5S,6S)-6-[(lR)-1-hydroxyethyl] -2-[(3S,5S)-5-(2-ethoxycarbonyl-1-moxy(hydroxy)iminoethyl)pyrrolidine-3-ylthio]-1-methylcarbapen-2-em-3-carboxylic acids (10a,l0b) were described. Methyl(2S,4S)-4-tritylthio-1-(allyloxycarbonyl)pyrrolidine-2-carboxyla late (I) was prepared from trans-4-hydroxy-L-proline with (2S,4R)-abs olute configuration as starting material. (2S,4S)-1-allyloxycarbonyl-2-(2-ethoxycarbonyl-hydroxy(methoxy )iminoethyl)-4-mercapto- pyrrolidines (6,7) were obtained from the tritylthio compound (I). (lR,5S,6S)-6-[(lR)-1-hydroxyethyl]-2-[(3S,5S)-5-(2-ethoxycarbonyl-1-methoxy(hydroxy)imino-ethyl)pyrrolidine-3-ylthio]-1-methylcarbapem-2-em-3-carboxylic acids (10a,10b) were obtained by the coupling reaction with carbapenem diphenylphosphates (8) and pyrrolidine-thiol moiety (6,7). Their in vitro antibacterial activities against both Gram-positive and Gram-negative were tested. Compounds ( 10a,10b) showed potent antibacterial activity except pseudomonas aerusinosa.

Identification of a Polyketide Synthase Gene in the Synthesis of Phleichrome of the Phytopathogenic Fungus Cladosporium phlei

  • So, Kum-Kang;Chung, Yun-Jo;Kim, Jung-Mi;Kim, Beom-Tae;Park, Seung-Moon;Kim, Dae-Hyuk
    • Molecules and Cells
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    • v.38 no.12
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    • pp.1105-1110
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    • 2015
  • Phleichrome, a pigment produced by the phytopathogenic fungus Cladosporium phlei, is a fungal perylenequinone whose photodynamic activity has been studied intensively. To determine the biological function of phleichrome and to engineer a strain with enhanced production of phleichrome, we identified the gene responsible for the synthesis of phleichrome. Structural comparison of phleichrome with other fungal perylenequinones suggested that phleichrome is synthesized via polyketide pathway. We recently identified four different polyketide synthase (PKS) genes encompassing three major clades of fungal PKSs that differ with respect to reducing conditions for the polyketide product. Based on in silico analysis of cloned genes, we hypothesized that the non-reducing PKS gene, Cppks1, is involved in phleichrome biosynthesis. Increased accumulation of Cppks1 transcript was observed in response to supplementation with the application of synthetic inducer cyclo-(${_L}-Pro-{_L}-Phe$). In addition, heterologous expression of the Cppks1 gene in Cryphonectria parasitica resulted in the production of phleichrome. These results provide convincing evidence that the Cppks1 gene is responsible for the biosynthesis of phleichrome.

Recombinant Human L-ferritin from Saccharomyces cerevisiae: Molecular Characterization and Synthesis of Iron Oxide Nanoparticles (효모에서 생산한 재조합 human L-ferritin의 생화학적 특성 및 나노입자의 철산화물 합성)

  • Kim, Kyung-Suk
    • KSBB Journal
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    • v.26 no.2
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    • pp.119-125
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    • 2011
  • In the synthesis of nanoparticles, much attention has been paid to regulating the particle size. There has been a possible evident that using the central cavity (core) of the protein ferritin has a greatly significant influence on it because the core can generate the nanometer-sized mineral particles of variable metal ions. In this report, recombinant human L-ferritins produced from Saccharomyces cerevisiae were purified and their molecular properties were characterized. The cDNA for human ferritin L chain was also expressed in another host such as Escherichia coli, and the properties of recombinant L-ferritins were compared. From isoelectric focusing experiment, the L-ferritin from the recombinant yeast showed no indication of N-glycosylation. Some post-translational modifications other than N-glycosylation were speculated in the L-ferritins from yeast. A difference was made in the L-ferritins in their iron uptake rates and the initial rate of the L-ferritin from yeast was slightly increased. The reconstitution yield and size distribution of the core minerals were analyzed in the L-ferritins by transmission electron microscopy. The L-ferritin from yeast with higher reconstitution yield (54.5%) showed slightly larger sizes (mean 6.92 nm) with narrower size distribution than the L-ferritin from E. coli. It is, in conclusion, speculated that L-ferritin from yeast is relatively superior to the other, in view of the size of nanoparticle and its relative homogeneity.

Biochemical Characterization of a Glycosyltransferase Homolog from an Oral Pathogen Fusobacterium nucleatum as a Human Glycan-Modifying Enzyme

  • Kim, Seong-Hun;Oh, Doo-Byoung;Kwon, Oh-Suk;Jung, Jae-Kap;Lee, Yun-Mi;Ko, Ki-Sung;Ko, Jeong-Heon;Kang, Hyun-Ah
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.859-865
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    • 2008
  • Bacterial glycosyltransferases have drawn growing attention as economical enzymes for oligosaccharide synthesis, with their easy expression and relatively broad substrate specificity. Here, we characterized a glycosyltransferase homolog (Fnu_GT) from a human oral pathogen, Fusobacterium nucleatum. Bioinformatic analysis showed that Fnu_GT belongs to the glycosyltransferases family II. The recombinant Fnu_GT (rFnu_GT) expressed in Escherichia coli displayed the highest glycosylation activity when UDP-galactose (Gal) was used as a donor nucleotide-sugar with heptose or N-acetylglucosamine (GlcNAc) as an acceptor sugar. Interestingly, rFnu_GT transferred the galactose moiety of UDP-Gal to a nonreducing terminal GlcNAc attached to the trimannosyl core glycan, indicating its potential as an enzyme for human-type N-glycan synthesis.

Synthesis of Methylated Anthranilate Derivatives Using Engineered Strains of Escherichia coli

  • Lee, Hye Lim;Kim, Song-Yi;Kim, Eun Ji;Han, Da Ye;Kim, Bong-Gyu;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.29 no.6
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    • pp.839-844
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    • 2019
  • Anthranilate derivatives have been used as flavoring and fragrant agents for a long time. Recently, these compounds are gaining attention due to new biological functions including antinociceptive and analgesic activities. Three anthranilate derivatives, N-methylanthranilate, methyl anthranilate, and methyl N-methylanthranilate were synthesized using metabolically engineered stains of Escherichia coli. NMT encoding N-methyltransferase from Ruta graveolens, AMAT encoding anthraniloyl-coenzyme A (CoA):methanol acyltransferase from Vitis labrusca, and pqsA encoding anthranilate coenzyme A ligase from Pseudomonas aeruginosa were cloned and E. coli strains harboring these genes were used to synthesize the three desired compounds. E. coli mutants (metJ, trpD, tyrR mutants), which provide more anthranilate and/or S-adenosyl methionine, were used to increase the production of the synthesized compounds. MS/MS analysis was used to determine the structure of the products. Approximately, $185.3{\mu}M$ N-methylanthranilate and $95.2{\mu}M$ methyl N-methylanthranilate were synthesized. This is the first report about the synthesis of anthranilate derivatives in E. coli.

Synthesis of Self-Assembled Peptide Nanoparticles Based on Dityrosine Covalent Bonds (다이타이로신 공유결합으로 자기조립된 펩타이드 나노입자의 합성)

  • Hur, Yun-Mi;Min, Kyoung-Ik
    • Korean Chemical Engineering Research
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    • v.59 no.1
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    • pp.112-117
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    • 2021
  • In this study, a method of self-assembly of peptides based on irreversible covalent bonds was studied by mimicking a biological covalent bond, dityrosine bond. A tyrosine-rich short peptide monomer having the sequence of Tyr-Tyr-Leu-Tyr-Tyr (YYLYY) was selected to achieve a high-density of dityrosine bond. The peptide nanoparticles covalently self-assembled with dityrosine bonds were synthesized by one-step photo-crosslinking of a peptide using a ruthenium catalyst under visible light. The effect of the concentration of each component for the size of the peptide nanoparticle was studied using dynamic light scattering, UV-Vis spectroscopy, and transmission electron microscopy. As a result, the synthesis conditions for size of the peptide nanoparticles ranging from 130 nm to 350 nm were optimized.

Enzymatic Synthesis of Resveratrol α-Glucoside by Amylosucrase of Deinococcus geothermalis

  • Moon, Keumok;Lee, Seola;Park, Hyunsu;Cha, Jaeho
    • Journal of Microbiology and Biotechnology
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    • v.31 no.12
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    • pp.1692-1700
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    • 2021
  • Glycosylation of resveratrol was carried out by using the amylosucrase of Deinococcus geothermalis, and the glycosylated products were tested for their solubility, chemical stability, and biological activities. We synthesized and identified these two major glycosylated products as resveratrol-4'-O-α-glucoside and resveratrol-3-O-α-glucoside by nuclear magnetic resonance analysis with a ratio of 5:1. The water solubilities of the two resveratrol-α-glucoside isomers (α-piceid isomers) were approximately 3.6 and 13.5 times higher than that of β-piceid and resveratrol, respectively, and they were also highly stable in buffered solutions. The antioxidant activity of the α-piceid isomers, examined by radical scavenging capability, showed it to be initially lower than that of resveratrol, but as time passed, the α-piceid isomers' activity reached a level similar to that of resveratrol. The α-piceid isomers also showed better inhibitory activity against tyrosinase and melanin synthesis in B16F10 melanoma cells than β-piceid. The cellular uptake of the α-piceid isomers, which was assessed by ultra-performance liquid chromatography (UPLC) analysis of the cell-free extracts of B16F10 melanoma cells, demonstrated that the glycosylated form of resveratrol was gradually converted to resveratrol inside the cells. These results indicate that the enzymatic glycosylation of resveratrol could be a useful method for enhancing the bioavailability of resveratrol.

The combination effects of PDGF and IGF-I on the proliferation and cellular activity of periodontal ligament cells (PDGF와 IGF-I 병용 사용시 치주인대세포의 증식과 세포활성에 미치는 영향에 관한 연구)

  • Suh, Jo-Young;Shin, Hong-In;Kyung, Hee-Moon
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.396-413
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    • 1996
  • Current acceptable methods for promoting periodontal regeneration are based on removal of diseased soft tissue. root treatment, guided tissue regeneration, introduction of new graft materials and biological mediators. Insulin-like growth factor-I(IGF-I) and Platelet-derived growth factor-BB(PDGF-BB), the members of the polypeptuyde growth factor family have been reported as the biological mediators which regulate a variety cellular matrix biologic activities of wound healing process including the cell proliferation, migration and extracellular matrix synthesis.The purposes of this study is to evaluate the combination effects of IGF-I and PDGF-BB on the cellular activity of the periodontal ligament cells to act as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM containing 10% FBS at the $37^{\circ}C$, 5% CO2 incubator. Author measured the DNA synthetic activity, and total protein, collagen and noncollagenous protein synthetic activities according to the concentration of 10,100ng/ml IGF-I and1,10 ng/ml PDGF-BB in combination. The results were as follows: Significantly increased in the 1 ng/ml PDGF-BB alone compared to the 10 ng/ml PDGF-BB alone(P<0.01) and in the 1 ng/ml PDGF-BB and 10, 100ng/ml IGF-I in combination compared to the 1 ng/ml PDGF-BB alone(P<0.05, P<0.0l). The synthetic activity of the total protein and collagen is significantly increased like to the synthetic activity of the DNA(P<0.05). The synthetic activity of the noncollagenous protein is increased according to the concentration of IGF_I, but not statistically statistically significant(P>0.05). The percent of the collagen is significantly in the 1ng/ml PDGF-BB and 10ng/ml IGF-I in combination compared to the 1ng/ml PDGF-BB alone(P<0.05) and in the 10ng/ml IGF-I in combination compared to the 10ng/ml PDGF-BB alone(P<0.05). The synthetic activity of the DNA is In conclusions, the percent study shows that PDGF-BB and IGF-I in combination have a potentiality to enhance the DNA synthesis and the total protein and collagen synthesis of The periodontal ligament cells, especially it is more significant in the low concentration of PDGF-BB compared to the high one. Thus, the PDGF-BB and IGF-I in combination may have important roles in promotion of periodontal litgment healing, and consequently, may useful for clinical application in periodontal regenerative procedures.

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Effects of Luteolin-7-𝑂-glucoside on melanin synthesis (Luteolin-7-𝑂-glucoside가 멜라닌 합성에 미치는 영향)

  • Choi, Byeong Min;Hong, Hyehyun;Park, Taejin;Kim, Seung-Young
    • Journal of Applied Biological Chemistry
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    • v.65 no.3
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    • pp.231-237
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    • 2022
  • Biorenovation is a method that converts existing compounds into new compounds through the enzymatic action of microorganisms. Biorenovation has expected effects such as reducing toxicity of compounds and increasing their activity. In this study, we successfully synthesized Luteolin-7-O-glucoside (L7G) through biorenovation and investigated its inhibitory effect on melanin production in α-Melanocyte stimulating hormone induced B16F10 mouse melanoma cells. We confirmed that Luteolin was toxic at 50, 100 and 200 µM, but our L7G in same concentration was not toxic for B16F10 mouse melanoma cells and also showed significant reduction in melanin production and tyrosinase activity. In addition, while investigating the effect of L7G on factors involved in melanin synthesis through western blotting, we were able to confirm that the MITF and tyrosinase protein synthesis was inhibited in treatment with L7G, however, tyrosinase related protein-1 (TRP-1) and dopachrome tautomerase (TRP-2) expression was not affected. So we derived a conclusion that through biorenovation we could produce compounds like L7G with improved activity and reduced toxicity for possible use as an active ingredient with whitening functionality in cosmetics.It also suggests that the application of biorenovation has potential usefulness in developing anti-inflammatory materials. It also suggests that the application of bio-renovation has potential usefulness in the development of inflammatory material. We applied Biorenovation technology to Distylium racemosum extract (DR) to generate Distylium racemosum biorenovation product (DRB), and investigated the anti-inflammatory properties of DRB in lipopolysaccharide (LPS)-treated RAW264.7 macrophages. We are applying technology to Biorenovation Distylium racemosum extract (DR) Distylium racemosum was to create a biorenovation product (DRB), lipopolysaccharide (LPS) investigated the anti-inflammatory properties of DRB in RAW264.7 macrophages treated for.

Synthesis of Sludge Waste-derived Semiconductor Grade Uniform Colloidal Silica Nanoparticles and Their CMP Application (슬러지 폐기물을 활용한 반도체급 균일한 콜로이달 실리카 나노입자의 제조 및 CMP 응용)

  • Kim, Dong Hyun;Kim, Jiwon;Jekal, Suk;Kim, Min Jeong;Kim, Ha-Yeong;Kim, Min Sang;Kim, Sang-Chun;Park, Seon-Young;Yoon, Chang-Min
    • Journal of the Korea Organic Resources Recycling Association
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    • v.30 no.3
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    • pp.5-12
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    • 2022
  • This study suggests the effective recycling method of sludge waste from various industrial fields to synthesize uniform colloidal silica nanoparticles. In detail, polymers are removed from the sludge waste to attain sludge-extracted silica (s-SiO2) micron-sized particles, and ammonia assisted sonication is applied to s-SiO2, which has effectively extracted the silanol precursor. The nano-sized silica (n-SiO2) particles are successfully synthesized by a typical sol-gel method using silanol precursor. Also, the yield amounts of n-SiO2 are determined by the function of s-SiO2 etching time. Finally, n-SiO2-based slurry is synthesized for the practical CMP application. As a result, rough-surfaced semiconductor chip is successfully polished by the n-SiO2-based slurry to exhibit the mirror-like clean surface. In this regard, sludge wastes are successfully prepared as valuable semicondutor grade materials.