• Title/Summary/Keyword: endo-B-1

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Nucleotide Sequence of Cellulolytic Xylanase Gene (bglBC2) from Bacillus circulans (Bacillus circulans 유래 cellulolytic xylanase 유전자(bglBC2)의 염기서열 결정 및 분석)

  • Kim, Ji-Yeon
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.67-72
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    • 2006
  • The nucleotide sequence of the cloned cellulolytic xylanase gene (bglBC2) from B. circulans ATCC21367 was determined. bglBC2 consists of an 1,224 bp open reading frame (ORF) coding for a polypeptide of 407 amino acids with a deduced molecular weight of 45 kDa. The Shine-Dalgarno (SD) sequence (5'-AAAGGAG-3') was found 9 bp upstream of the initiation codon, ATG. A promoter region corresponding closely to the B. subtilis consensus sequence (-35: TTGACA,-10: TATAAT) was detected, the putative -35 and -10 sequences of which were TTTACA and TATACT, respectively. The deduced amino acid sequence of the cellulolytic xylanase showed 97% homology with that of the alkaline $endo-\beta-1,4-glucanase$ from B. circulans KSM-N257, 75% homology with that of the $endo-\beta-1,3-1,4-glucanase$ from B. circulans WL-12, and 45% homology with that of the $endo-\beta-1,4-glucanase$ (cellulase) from Bacillus sp. KSM-330. The bglBC2 sequence was deposited in Gen-Bank under the accession number AY269256.

Production of Endo-Polygalacturonase of a Mutant of Aspergillus niger (Aspergillus niger의 변이주(變異株)에 의(依)한 Endo-polygalacturonase의 생산(生産))

  • Park, Yoon Joong;Shon, Cheon Bae
    • Korean Journal of Agricultural Science
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    • v.12 no.2
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    • pp.324-332
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    • 1985
  • Aspergillus niger B-15 with strong Endo-polygalacturonase (Endo-PG) activities was selected out from a total of 1,573 fungal strains isolated from various testing materials. A mutant strain, U-46, was obtained from the Aspergillus niger B-15 by repeated irradition of ultra-violet light. The objectives of the study were to investigate the fungal properties of the parental and mutant strains obtained and to study the condition of enzyme production and reaction. The results obtained are summarized as follows: 1. The size of conidial head of the U-46 mutant was smaller than that of the parental strains, B-15 and the length of the conidiophore was also shorter than that of the parental strains. 2. The optimum conditions for the Endo-PG production of the parental B-15 strain in the wheat bran Koji were obtained when 40% of water was added to the wheat bran and the temperature was 30 to $35^{\circ}C$. However, the best condition for the mutant U-46 strain was attained when 60 to 70% of water was added and the temperature was $35^{\circ}C$. The optimum growing periods were two to three days for both parental and mutant strains. 3. Under the optimum producing conditions of each strains, the enzymatic activity of the mutant U-46 was 20 times higher than the Endo-PG of the parental strain, B-15. 4. When both strains were cultured in the wheat bran Koji containing 60% of water at $35^{\circ}C$ for three days, the mutant strain. U-46, was about 46 times higher in the Endo-PG activity and about 18 times greater in Exo-PG activity than the parental strain, B-15. The activities of cellulase, $\alpha$-amylase, and glucoamylase were also highly increased in the mutant strain. 5. The mutant strain, U-46, increased its Endo-PG activity up to 20% over that of ordinary case when 1.2 to 1.5% of ammonium sulphate was added to the wheat bran. 6. The optimum condition for Endo-PG activity of crude enzyme of the mutant strain, U-46, was attained when pH of reaction solution was 4.0 to 4.5 and the temperature was $50^{\circ}C$.

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Characterization of the 5-Flanking region upstream from the structural gene for Zymononas mobilis alcohol dehydrogenase

  • Yoon, Ki-Hong;Park, Seung-Hwan;Jung, Kyung-Hwa;Pack, M. Y.
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.126-127
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    • 1995
  • A Zymomonas mobilis DNA fragment consisting of 207 nucleotides, which corresponded to the 5'-flanking region of an adhB gene encoding alcohol dehydrogenase II, was fused to the structural gene coding for a Bacillus endo-.betha.--1, 4-glucanase. The Z. mobilis DNA framgment waw identified to promote 50-fold increase in the expression of endo-.betha.1. 4 glucanase gene in Escherichia coli.

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Molecular Cloning and Nucleotide Sequence of Endo-Inulinase Gene from Xanthomonas oryzae #5

  • Kim, Byeong-U;Kim, Mi-Rang;Yu, Dong-Ju
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.655-659
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    • 2000
  • A 11.5-kb DNA fragment containing an endo-inulinase gene was cloned from Xanthomonas oryzae #5. It contained a single open reading frame of 3,999bp, encoding a polypeptide composed of signal peptide of 32 amino acids and mature protein of 1,301 amino acids. From the comparison of amino acids sequences with fructan hydrolases, inulinase, levanase and CFTase, the sequence of the endo-inulinase had highly homology of 72% with CFTase of B. circulans, and six highly conserved regions including the ${\beta}-fructosidase$ motif were found.

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Endo-1,4-β-xylanase B from Aspergillus cf. niger BCC14405 Isolated in Thailand: Purification, Characterization and Gene Isolation

  • Krisana, Asano;Rutchadaporng, Sriprang;Jarupan, Gobsuk;Lily, Eurwilaichitr;Sutipa, Tanapongpipat;Kanyawim, Kirtikara
    • BMB Reports
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    • v.38 no.1
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    • pp.17-23
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    • 2005
  • During the screening of xylanolytic enzymes from locally isolated fungi, one strain BCC14405, exhibited high enzyme activity with thermostability. This fugal strain was identified as Aspergillus cf. niger based on its morphological characteristics and internal transcribed spacer (ITS) sequences. An enzyme with xylanolytic activity from BCC14405 was later purified and characterized. It was found to have a molecular mass of ca. 21 kDa, an optimal pH of 5.0, and an optimal temperature of $55^{\circ}C$. When tested using xylan from birchwood, it showed $K_m$ and $V_{max}$ values of 8.9 mg/ml and 11,100 U/mg, respectively. The enzyme was inhibited by $CuSO_4$, EDTA, and by $FeSO_4$. The homology of the 20-residue N-terminal protein sequence showed that the enzyme was an endo-1,4-$\beta$-xylanase. The full-length gene encoding endo-1,4-$\beta$-xylanase from BCC14405 was obtained by PCR amplification of its cDNA. The gene contained an open reading frame of 678 bp, encoding a 225 amino acid protein, which was identical to the endo-1,4-$\^{a}$-xylanase B previously identified in A. niger.

Comparison of apical sealing ability of continuous wave of obturation technique using EndoTwinn and System B (EndoTwinn과 System B continuous wave of obturation units를 이용한 근단부 근관충전 효율비교)

  • Shin, Hyun-Ju;Park, Jeong-Kil;Hur, Bock;Kim, Hyeon-Cheol
    • Restorative Dentistry and Endodontics
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    • v.32 no.6
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    • pp.522-529
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    • 2007
  • The purpose of this study was to compare the apical leakage of the root canal filled with the System B and the EndoTwinn(the combined application of heat and ultrasonic vibration). Sixty extracted premolars with straight root were cleaned and shaped to size 35. Group SB was obturated using System B and Group ET was filled with EndoTwinn. A size 35 of 0.06 tapered gutta-percha and Adseal were used and the plugger which could be introduced to 4 mm short of working length was selected in the obturation procedure. As the positive control, Group PC was not filled. In Group SB, ET and PC, all external surfaces of each tooth were coated with nail varnish leaving only 1 mm area around the apical foramen. In the negative control of Group NSB and Group NET, all of external tooth surface including apical foramen was coated with the nail varnish. The specimens were immersed in methylene blue dye solution for 2 days. Then the specimens were sectioned at each 1 mm from apex to 5 mm level. The final score of one specimen was given by summing up of the points at all levels. The dye leakage of Group ET was significantly less than that observed in Group SB (p < 0.05). And the frequency of gutta-percha pulling out from root canal when the plugger was removed was more often with the System B than with EndoTwinn but there was no significant difference.

DFT Study of p-tert-Butylcalix[5]crown-6-ether Complexed with Alkylammonium Ions

  • Oh, Dong-Suk;Choe, Jong-In
    • Bulletin of the Korean Chemical Society
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    • v.28 no.4
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    • pp.596-600
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    • 2007
  • The structures and energies of p-tert-butylcalix[5]crown-6-ether (1) and its alkylammonium complexes have been calculated by DFT B3LYP/6-31G(d,p) method. We have studied the binding sites of these host-guest complexes focusing on the p-tert-butylcalix[5]arene pocket (endo) or the crown-6-ether moiety (exo) of 1. The smaller alkylammonium cations have the better complexation efficiency with p-tert-butylcalix[5]crown-6- ether than the bulkier alkylammonium ions. For the sec- and tert-butylammonium ions, the hydrogen-bond distances of the exo-complexes are shorter, therefore, stronger than the endo-cases. This DFT calculated result is in parallel with the trend of the experimental association constants of the branched butylammonium ions.

Expression and Secretion of Foreign Proteins in Yeast Using the ADH1 Promoter and 97 K Killer Toxin Signal Sequence

  • Hong, Seok-Jong;Kang, Hyen-Sam
    • BMB Reports
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    • v.31 no.2
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    • pp.123-129
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    • 1998
  • Foreign proteins, $endo-{\beta}-1,4-glucanase$ of Bacillus subtilis, preS1+S2 region of hepatitis B virus large surface antigen, human ${\beta}_2-adrenergic$ receptor ($h{\beta}_{2}AR$), and bovine growth hormone (bGH) were expressed in Saccharomyces cerevisiae and secreted into the medium. These proteins were expressed using the alcohol dehydrogenase I (ADH1) promoter of Saccharomyces cerevisiae and secreted by signal sequence of the 97 K killer toxin gene of doublestranded linear DNA plasmid (pGKL1) of S. cerevisiae. All these proteins underwent severe modifications; in particular, N-glycosylation in the case of $endo-{\beta}-1,4-glucanase$, $h{\beta}_2AR$, and preS1+S2. Seventy four percent of the expressed $endo-{\beta}-1,4-glucanase$ was secreted into the culture medium. Highly modified proteins were detected in the culture medium and in the cell. Expressed $h{\beta}_2AR$, which has seven transmembrane domains, remained in the cell. The degrees of secretion and modification and the states of proteins in the culture medium and in the cell were quite different. These results indicated that the nature of the protein has a critical role in its secretion and modifications.

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Synthesis of Octahydro-2,3-dioxo-cyclopenta[b]pyrrole-3a-carboxylates by Nucleophilic Addition to N-Acyliminium (N-Acyliminium에의 친핵성 부가에 의한 Octahydro-2,3-dioxo-cyclopenta[b]pyrrole-3a-carboxylates의 합성)

  • Seo Won-Jun;Chang-hee Jung;Seung-Ju Choi;Young-Kyu Park;Tae-Heung Kim;Sang-Kyu Lee
    • Journal of the Korean Chemical Society
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    • v.38 no.12
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    • pp.908-914
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    • 1994
  • 6a-Hydroxy- and 6a-methoxy-derivatives (1∼6) of octahydro-2,3-dioxo-cyclopenta[b]-pyrrole-3a-carboxylate were synthesized from oxalylation of enamine A, which was prepared from condensation of five-membered cyclic ${\beta}$-keto ester and phenylalkylamine, followed by addition of water or methanol. The formation of heterocyclic ring was assumed to occur by the way of unstable N-acyliminium (B). Stable adduct C (1∼6) was obtained from nucleophilic addition to the endo-ene type pyrrolinium B.

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The Endo-β-1,4-Glucanase of Bacillus amyloliquefaciens Is Required for Optimum Endophytic Colonization of Plants

  • Fan, Xiaojing;Yang, Ruixian;Qiu, Sixin;Cai, Xueqing;Zou, Huasong;Hu, Fangping
    • Journal of Microbiology and Biotechnology
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    • v.26 no.5
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    • pp.946-952
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    • 2016
  • The eglS gene in Bacillus amyloliquefaciens encodes an endo-β-1,4-glucanase that belongs to glycosyl hydrolase family 5. In this study, a disruption mutant of gene eglS was constructed to examine its role in bacterial adaptation in plants. The mutant TB2k, eglS gene inactivated bacterial strain, was remarkably impaired in extracellular cellulase activity. When inoculated on Brassica campestris, the TB2k population was reduced by more than 60% compared with the wild-type strain in the root, stem, and leaf tissues. Overexpression of eglS in the wild-type strain increased the bacteria population in the plant tissues. Further studies revealed that the transcription level of eglS was correlated with bacterial population. These data demonstrate that endo-β-1,4-glucanase of B. amyloliquefaciens is required for its optimal endophytic colonization.