• Title/Summary/Keyword: Molecular Characteristics

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Molecular Cloning and Characterization of Expression Patterns of a Plastid ω-3 Fatty Acid Desaturase cDNA from Perilla frutescens

  • Lee, Seong-Kon;Kim, Kyung-Hwan;Kwon, Moo-Sik;Hwang, Young-Soo
    • Journal of Applied Biological Chemistry
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    • v.44 no.1
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    • pp.6-11
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    • 2001
  • An ${\omega}-3$ fatty acid desaturase gene which is involved in de novo synthesis of -Iinolenate was isolated from cDNA library of Perilla frutescens. A cDNA library was constructed with mRNA extracted from perilla seeds of 12 DAF. The cDNA clone consisting of 1317-bp open reading frame encoding 438 amino acids with a relative MW of 50kDa, was isolated and showed 65-83% similarities to other known genes. This cDNA is deduced to encode a plastidal ${\omega}-3$ fatty acid desaturase based on the fact that it has higher homology to plastidal ones than to microsomal ones and its N-terminal sequence shares several characteristics of transit peptides of chloroplast proteins. Southern blot analysis of genomic DNA indicated that more than one gene or alleles for ${\omega}-3$ fatty acid desaturase are present in the genome of perilla. Northern blot analysis showed that the ${\omega}-3$ fatty acid desaturase gene is mainly revealed in early developing seeds and has different expression patterns depending on tissue types compared to the microsomal ones.

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Function of Cell-Bound and Released Exopolysaccharides Produced by Lactobacillus rhamnosus ATCC 9595

  • Kim Ji-Uk;Kim Young-Hoon;Han Kyoung-Sik;Oh Se-Jong;Whang Kwang-Youn;Kim Jai-Neung;Kim Sae-Hun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.939-945
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    • 2006
  • The physiological characteristics and function of the exopolysaccharide (EPS) produced by Lactobacillus rhamnosus ATCC 9595 were determined. The total quantity of EPS was rapidly increased to 496$\pm$20 mg/l during the exponential phase, and then maintained steadily during the stationary phase. During the exponential phase (18 h), the total EPS consisted of 61% cell-bound EPS (cb-EPS) and 39% released EPS (r-EPS), whereas the relative proportion of EPS during the stationary phase (48 h) was convered to 23% cb-EPS and 77% r-EPS. On gel permeation chromatography, cb-EPS was fractionated as a single peak of 8.6$\times10^6$ Da, whereas r-EPS was fractionated as two peaks with average molecular weights of 4.3$\times$10$^4$ and 8.6$\times10^6$ Da. Interestingly, both EPS species exhibited anticancer properties and cholera toxin-binding activities. Our results suggest that the EPS generated by L. rhamnosus ATCC 9595 might be suitable for use as a functional food or food supplement.

Characterization of Bacillus thuringiensis StrainBT-14 having Insecticidal Activity against Plutella xylostella

  • Bok, Song-Hae-Jung, Yong-Chul;Kim, Sung-Uk;Son, Kwang-Hee;Lee, Hyung-Hoan
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.322-326
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    • 1994
  • Bacillus thuringiensis strain BT-14 was isolated from alfalfa dust in Korea. The strain BT-14 produced one bipyramidal crystal and one spore in the cell. The biochemical characteristics of the strain BT-14 were similar to those of Bacillus thuringiensis subsp. kurstaki HD-l. Examination of its antibiotic resistance revealed that while the strain BT-14 was less resistant than BTK HD-l to ampicillin, gentamycin, neomycin and tobramycin, it was more resistant to amikacin than BTK HD-l. The $\delta$-endotoxin crystal of strain BT-14 consisted of a single protein with a high molecular weight of ca 135 KD on a 10% SDS-PACE. The strain BT-14 contained at least nine different plasmids with sizes of 2.9, 5.3, 5.8, 6.2, 9.4, 15.1, 18.1, 23.1 and 79 Kb. In insect bioassay, the isolated strain BT-14 showed lethality of 67% against Plutella xylostella larvae at dilution of 5$\times$$l0^{-4}$ (5$\times$l0 to 3$\times$$l0^2$ spores/ml), which is, almost equivalent to that of BTK HD-l.

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Metarizium anisopliae (Metschn.) Sorok이 생산하는 Lactobacillus plantarum 용균효소의 분리, 정제 및 특성

  • Ryoo, Ky-Chul;Hahm, Byoung-Kwon;Paik, Un-Wha;Yu, Ju-Hyun;Bai, Dong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.24 no.6
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    • pp.678-686
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    • 1996
  • To improve the preservation of Kimchi, we isolated Lactobacillus plantarum lytic enzyme-producing strain from soil, and the enzyme was purified and characterized. From the observation of cultural and morpho- logical characteristics, the isolated strain was identified as Metarrisium anisopliae (Metschn.) Sorok. The enzyme was purified to 75-folds with 40% yields through affinity adsorption and CM-Sephadex C-50 column chromatog- raphy. The optimum pH and temperature for lytic activity are 4.0 and 40$\circ$C, respectively, and the enzyme acitvity is stable between pH 3.0 and 9.0, and up to 50$\circ$C. The enzyme is a monomeric protein with molecular weight of 40,000 daltons by SDS-PAGE and gel filtration. The enzyme is endopeptidase which breaks the peptide linkage of Lactobacillus plantarum peptidoglycan. The lytic action spectra confirmed that Leuconostoc mesenteroides, a useful strain for the fermentation of Kimchi, is not lysed by the enzyme. The enzyme activity is inhibited by N-bromosuccinimide (NBS), which probably indicates the involvement of tryptophan residue in active site of the enzyme, and also inhibited by Ag$^{+}$. The amino acid composition analysis showed that the enzyme contains more acidic amino acids than basic ones, and composition of alanine, glycine, proline and tyrosines was very high.

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Heterologous Expression of Recombinant Transglutaminase in Bacillus subtilis SCK6 with Optimized Signal Peptide and Codon, and Its Impact on Gelatin Properties

  • Wang, Shiting;Yang, Zhigang;Li, Zhenjiang;Tian, Yongqiang
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.1082-1091
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    • 2020
  • Microbial transglutaminases (MTGs) are widely used in the food industry. In this study, the MTG gene of Streptomyces sp. TYQ1024 was cloned and expressed in a food-grade bacterial strain, Bacillus subtilis SCK6. Extracellular activity of the MTG after codon and signal peptide (SP Ync M) optimization was 20 times that of the pre-optimized enzyme. After purification, the molecular weight of the MTG was 38 kDa and the specific activity was 63.75 U/mg. The optimal temperature and pH for the recombinant MTG activity were 50℃ and 8.0, respectively. MTG activity increased 1.42-fold in the presence of β-ME and 1.6-fold in the presence of DTT. Moreover, 18% sodium chloride still resulted in 83% enzyme activity, which showed good salt tolerance. Cross-linking gelatin with the MTG increased the strength of gelatin 1.67 times and increased the thermal denaturation temperature from 61.8 to 75.8℃. The MTG also significantly increased the strength and thermal stability of gelatin. These characteristics demonstrated the huge commercial potential of MTG, such as for applications in salted protein foods.

Preliminary Evidence for a Metallothionein-like Cd-binding Protein in the Kidney of the Antarctic Clam Laternula elliptica

  • Choi, Hee-Seon, J.;Ahn In-Young;Ryu Soo-Kyung;Lee, Yong-Seok;Lee, In-Sook;Jeong, Kye-Heon
    • Ocean and Polar Research
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    • v.23 no.4
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    • pp.337-345
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    • 2001
  • A Cd-binding protein was identified in the renal cytosol of the Antarctic clam Laternula elliptica which naturally contains high concentrations of Cd. The Cd-binding protein showed similar characteristics of metallothionein (MT) in molecular weight (about 10-12 kDa) and low spectral absorbance at 280 nm with relatively high absorbance at 254nm. Results of immuno-histochemical staining suggested that the MT-like Cd-binding protein was mainly located in the epithelial cells of the kidney. The MT-like protein was a major ligand of cytosolic Cd as shown in the elution profiles of chromatography and may play an important role in Cd sequestration and accumulation in L. elliptica kidney. A considerable amount of Cd was also found to be associated with particulate fraction, indicating the sequestration to particulate fraction is as important as binding to the cytosolic MT-like protein in Cd accumulation in the kidney.

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Synthesis of Benzoic Acid Modified Polyester Containing Phosphorus for Flame-Retardant Coatings (난연도료용 인 함유 벤조산 변성폴리에스테르의 합성)

  • Chung, Dong-Jin;Lee, Ae-Ri;You, Hyuk-Jae;Jung, Choong-Ho;Park, Hong-Soo
    • Journal of the Korean Applied Science and Technology
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    • v.21 no.1
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    • pp.51-61
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    • 2004
  • Reaction intermediates PCP/BZA (PBI) and tetramethylene bis(orthophosphate) (TBOP) wer synthesized from polycaprolactone (PCP) and benzoic acid (BZA) and from pyrophosphoric acid and 1,4-butanediol, respectively. Benzoic acid modified polyesters containing phosphorus (APTB-5, -10, -15) were synthesized by polycondensation of the prepared PBI (containing 5, 10, 15wt% of benzoic acid), TBOP, adipic acid, and 1,4-butanediol. The structure and characteristics of APTBs were examined using FT-IR, NMR, GPC, and TGA analysis. The increase of the amount of BZA in the synthesis of APTBs resulted in decrease in average molecular weight and kinematic viscosity. From the TGA analysis of APTBs, it was found that the afterglow decreased with the amount of BZA content at the high temperatures.

A Physicochemical Characteristics and Manufacture of Solubility Collagen Peptide from Flatfish Skin (당가자미 껍질로부터 수용성콜라겐 제조 및 이화학적 특성)

  • Jang, Boo-Sik;Lee, Mi-Jin;Jeong, Noh-Hee
    • Journal of the Korean Applied Science and Technology
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    • v.30 no.3
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    • pp.560-566
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    • 2013
  • In this research we extracted water-soluble collagen peptide from flatfish skin and compared it with commercially available collagen peptide extracted from Tilapia scale currently placed on the market in the aspect of physiochemical property. The physical property and nutritional components of FSCP appeared almost similarly to those of TSCP, and also in calorie, FSCP marking 3.82 Kcal showed no differences from TSCP marking 3.84 Kcal. As for forming amino acids, in aspartic acid, serine, histidine, tyrosine, methionine, FSCP had higher content than TSCP, but in OH-proline, proline and alanine FSCP had lower content than TSCP. Especially the content of essential amino acids of FSCP marked 22.74% with a higher content compared with 13.64% of TSCP. In the distribution of molecular weight FSCP with 1,000 Da showed comparatively low compared with TSCP, and in emulsion property and stability FSCP and TSCP showed similar excellent trend.

Effects of γ-Aminobutyric Acid (GABA)-Enriched Sea Tangle Laminaria japonica Extract on Lipopolysaccharide-Induced Inflammation in Mouse Macrophage (RAW 264.7) Cells

  • Choi, Ji-Il;Yun, In-Hye;Jung, Yeounjoong;Lee, Eun-Hye;Nam, Taek-Jeong;Kim, Young-Mog
    • Fisheries and Aquatic Sciences
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    • v.15 no.4
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    • pp.293-297
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    • 2012
  • ${\gamma}$-Aminobutyric acid-enriched sea tangle extract was obtained from the fermentation of Lactobacillus brevis BJ-20. The fermented sea tangle extract (FST) was separated into three fractions by molecular weight: FST I (greater than 10 kDa), FST II (1-10 kDa), and FST III (less than 1 kDa). The anti-inflammatory characteristics of the FST fractions were investigated by measuring the production of nitric oxide (NO) and the expression levels of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-induced mouse macrophage (RAW 264.7) cells. Both NO production and iNOS expression levels were significantly inhibited by FST treatments in a dose-dependent manner. FST III was the most effective inhibitor of processes. This demonstrates that the effect of FST on LPS-induced inflammation might be closely correlated with the inhibition of inflammatory cytokine expression.

Data Mining Techniques for Analyzing Promoter Sequences (프로모터 염기서열 분석을 위한 데이터 마이닝 기법)

  • 김정자;이도헌
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2000.10a
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    • pp.328-332
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    • 2000
  • As DNA sequences have been known through the Genome project the techniques for dealing with molecule-level gene information are being made researches briskly. It is also urgent to develop new computer algorithms for making databases and analyzing it efficiently considering the vastness of the information for known sequences. In this respect, this paper studies the association rule search algorithms for finding out the characteristics shown by means of the association between promoter sequences and genes, which is one of the important research areas in molecular biology. This paper treat biological data, while previous search algorithms used transaction data. So, we design a transformed association nile algorithm that covers data types and biological properties. These research results will contribute to reducing the time and the cost for biological experiments by minimizing their candidates.

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