• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.370-375
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• 2016

Two xylanase genes were cloned into Escherichia coli from Bacillus sp. YB-1401 and B. amyloliquefaciens YB-1402, which had been isolated as mannanase producer from home-made doenjang, respectively, and their nucleotide sequences were determined. Both xylanase genes consisted of 642 nucleotides, encoding polypeptides of 213 amino acid residues. The deduced amino acid sequences of the YB-1401 and YB-1402 xylanase, designated Xyn1401 and Xyn1402, differed from each other by single amino acid residue, Asn for Xyn1401 and Lys for Xyn1402, corresponding to amino acid position of 127. Their amino acid sequences were highly homologous to those of xylanases belonging to the glycosyl hydrolase family 11. The 28 amino acid stretch in the N-terminus of both enzymes was predicted as signal peptide by SignalP4.1 server. Both xylanases were localized at the level of 91−94% in culture filtrate of the recombinant E. coli cells, suggesting they were secreted efficiently in E. coli cells. The optimal reaction conditions were 50℃ and pH 6.0 for Xyn1401, and 55℃ and pH 6.5 for Xyn1402, respectively, indicating one amino acid difference from each other affected pH and temperature profiles of their activities. In addition, their thermostabilities were somewhat different from each other.

• Journal of Life Science
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• v.14 no.4
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• pp.636-643
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• 2004

D-Xylose isomerase produced by Lactococcus sp. JK-8, isolated from kimchi, was purified 17-fold of homogeneity, and its physicochemical properties were determined. Although the N-terminal amino acid sequence of D-xylose isomerase was analysed to Ala-Tyr-Phe-Asn-Asp-Ile-Ala-Pro-Ile-Lys, it was not similar to that of Lactobacillus enzyme. The molecular weight of the purified enzyme was estimated to be 180 kDa by gel filtration, 45 kDa by SDS-PAGE and the enzyme was homotetramer. The optimum pH of the enzyme was around 7 and stable between pH 6 and 8. The optimum reaction temperature was 7$0^{\circ}C$ and stable up to 7$0^{\circ}C$ in the presence of 1 mM $Mn^{2+}$. Like other D-xylose isomerases, this enzyme required divalent cation, such as $Mg^{2+}$, $Co^{2+}$, or $Mn^{2+}$ for the activity and thermostability. $Mn^{2+}$was the best activator. Substrate specificity studies showed that this enzyme was highly active on D-xylose. The enzyme had an isoelectric point of 4.8, and fm values for D-xylose was 5.9 mM.

• Journal of Marine Bioscience and Biotechnology
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• v.7 no.1
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• pp.19-27
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• 2015

The objective of the current study was to evaluate the inhibitory and antioxidant activities of powdered katsuobushi (dried bonito) protein hydrolysates and their corresponding fractions. The powdered katsuobushi (dried bonito) hydrolysates were obtained by enzymatic hydrolysis using Alcalase, ${\alpha}$-chymotrypsin, Neutrase, pepsin, papain, and trypsin. The antioxidant efficacy of the respective hydrolysates were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, superoxide, and alkyl radical-scavenging activities. Among the hydrolysates, the peptic-derived hydrolysate exhibited the highest antioxidant activity compared to other enzymatic hydrolysates. Therefore, the peptic-derived hydrolysate was further analyzed, and was found to contain an active peptide with an amino acid sequence identified as Pro-Met-Pro-Leu-Asn-Ser-Cys (756 Da). The purified peptides from powdered katsuobushi (dried bonito) had an $EC_{50}$ value of $105.82{\mu}M$, and exhibited an inhibitory effect against DNA oxidation induced by hydroxyl radicals. Taken together, these results suggests that powdered katsuobushi (dried bonito) could be used as a natural antioxidant in functional foods and prevent oxidation reactions in food processing.

• Journal of the Korean Magnetic Resonance Society
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• v.1 no.2
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• pp.112-125
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• 1997

It has been demonstrated that mutant Hemoglobins (Hb) which have an altered ${\alpha}$1${\beta}$2 subunit interface can be designed. A compensatory mutation for a naturally occurring abnormal human Hb, Hb Kempsey (${\beta}$99Asp\longrightarrowAsn), has been designed, and this mutation allowed the molecule to regain its allosteric response. The calculated values for the difference in the free energy of cooperativity show excellent agreement with experimentally determined thermodynamic values, suggesting that the molecular dynamics simulation results can be used to obtain information about the specific interactions which contribute to the total free energy of cooperativity. These results provide encouragement to begin a systematic investigation of the molecular basis of the subunit interactions between the ${\alpha}$1 and ${\beta}$2 chains of Hb A by designing appropriate r Hbs. These studies could lead to the design of Hbs with desired cooperativity in the oxygenation process and to the restoration of functional properties of abnormal hemoglobins associated with hemoglobinopathies. Thus, the present results also have the implications in using gene therapy to treat patients with hemoglobinpathies.

• The Korean Journal of Applied Statistics
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• v.18 no.1
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• pp.57-66
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• 2005

In this paper, we propose a method to evaluate the solutions of the renewal equations related to SPRT for Erlang distribution. In SPRT, the Average Sample Number(ASN) and type I or type II error probabilities are shown in Fredholm type integral equations. The integral equations are generally solved by the approximation method using Gaussian quadrature. For Erlang distribution, it has been known that the exact solutions of the equations exist. We propose the algorithm to solve the equations.

• BMB Reports
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• v.44 no.10
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• pp.653-658
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• 2011

Sclerotinia sclerotiorum fungus has three endoxylanases induced by wheat bran. In the first part, a partial xylanase sequence gene (90 bp) was isolated by PCR corresponding to catalytic domains (${\beta}5$ and ${\beta}6$ strands of this protein). The high homology of this sequence with xylanase of Botryotinia fuckeliana has permitted in the second part to amplify the XYN1 gene. Sequence analysis of DNA and cDNA revealed an ORF of 746 bp interrupted by a 65 bp intron, thus encoding a predicted protein of 226 amino acids. The mature enzyme (20.06 kDa), is coded by 188 amino acid (pI 9.26). XYN1 belongs to G/11 glycosyl hydrolases family with a conserved catalytic domain containing $E_{86}$ and $E_{178}$ residues. Bioinformatics analysis revealed that there was no Asn-X-Ser/Thr motif required for N-linked glycosylation in the deduced sequence however, five O-glycosylation sites could intervene in the different folding of xylanses isoforms and in their secretary pathway.

• Journal of KIISE:Information Networking
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• v.28 no.1
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• pp.45-55
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• 2001

CORBA 보안 서비스는 네트워크를 기반으로 하는 분산 환경 하에서 데이터 전송 시 사용자가 요구하는 수준의 비밀성 보장과 무결성 제공을 위해 비보호 무결성, 비밀성, 부결성 및 비밀성과 같은 파라미터를 갖는 QoP 기능을 지원하고 있다 그러나 기존의 QoP 기능은 전자상거래, 재무, 통신 CORBA Med와 같은 광범위한 CORBA의 응용 영역들간에 특정 암호화 알고리즘에 대한 서로 다른 정책을 가질 경우 전송되는 데이터에 대한 무결성과 비밀성을 지원할 수 없는 문제접을 갖고 있다. 이를 해결하기 위하여 본논문에서는 광범위한 CORBA의 응용영역들간에 데이터 전송 시 암호화 알고리즘 관리 및 제어기능을 개선한 QoP 모델을 제안한다. 개선된 QoP 관리 및 제어 모델은 OMG에서 발표한 RFP의 요구사항을 기반으로 설계되었으며 특정 암호화 알고리즘들의 관리와 실행중에 사용되는 암호화 알고리즘의 변경제어와 암/복호화시 비도 설정을 지원하다, 구현 결과 개선된 QoP 관리 및 제어 모델은 CORBA IDL 로 설계하여 개발자로 하여금 응용개발의 용이성을 부여하였으며 광범위한 CORBAdmd용 영역들간에 암호화 알고리즘에 대한 QoP 기능을 지원한다. 또한 ISO/IEC 8824(ASN. 1)에서 정의된 OID를 사용하여 암호화 알고리즘의 호환성을 제공한다.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.510-515
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• 2005

Pseudomonas cepacia BY21 was found to produce glutaryl acylase that is capable of deacylating glutaryl-7-aminocephalosporanic acid (glutaryl-7-ACA) to 7-aminocephalosporanic acid (7-ACA), which is a starting material for semi-synthetic cephalosporin antibiotics. Amino acids of the reported glutaryl acylases from various Pseudomonas sp. strains show a high similarity (>93% identity). Thus, with the known nucleotide sequences of Pseudomonas glutaryl acylases in GenBank, PCR primers were designed to clone a glutaryl acylase gene from P. cepacia BY21. The unknown -subunit gene of glutaryl acylase from chromosomal DNA of P. cepacia BY21 was cloned successfully by PCR. The -subunit amino acids of P. cepacia BY21 acylase (GenBank accession number AY948547) were similar to those of Pseudomonas diminuta KAC-1 acylase except that Asn408 of P. diuminuta KAC-1 acylase was changed to Leu408.

• Microbiology and Biotechnology Letters
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• v.28 no.4
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• pp.195-201
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• 2000

Glucoamylase of Saccharomyces diastaticus is produced as a large precursor composed of signal peptide (21 amino acid residues), Thr and Ser-rich region and functional glucoamylase. To evaluate the utility of the glucoamylase signal peptide (GSP) for the secretion of foreign proteins, four types of GSP mutants (ml : Pro-18 longrightarrowLeu-18, m2 : Tyr-13 longrightarrowLeu, m3 : Ser-9longrightarrowLeu-9, m4 : Asn-5 longrightarrowPro-5) were constructed and secretion efficiency of each mutant was compared with that of native GSP by the expression and secretion of Bacillus subtilis CMCase under the control of GAP in N-terminal domain and hydrophobic domain. n mutant 4, a polar amino acid was replaced by a helix - breaking Pro residue. CMCase activity assay and Western blot analysis revealed that CMCase secretion by GSP mutants replaced by Leu were increased compared with native GSP. In the case of m2 and m3, the substitution of Leu for Tyr-13 and Ser-9 in the hydrophobic region resulted in a twofold increase in the extracellular CMCase activity.

• Korean Journal of Environmental Agriculture
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• v.16 no.1
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• pp.55-60
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• 1997

Heavy metal-tolerant microorganisms, such as Pseudomonas putida, P. aeruginosa, P. chlororaphis and P. stutzeri which possessed the ability to accumulate cadmium, lead, zinc and copper, respectively, were isolated from industrial wastewaters and mine wastewaters polluted with various heavy metals. The distribution of heavy metal in the cell components, and amino acid compositions, was investigated. The distribution of heavy metal in the cell fractions of each heavy metal-tolerant microorganism grown for 20 hours in the basal medium containing 100mg/l of each heavy metal was investigated. In the case of cadmium-tolerant P. putida, lead-tolerant P. aeruginosa and copper-tolerant P. stutzeri, approximately $50{\sim}60%,\;30{\sim}40%$ and $10{\sim}17%$ of each heavy metal absorbed were distributed to cell wall, cell membrane and cytoplasm fractions, respectively. In the case of zinc-tolerant P. chlororaphis, approximately 32%, 55% and 13% of zinc were distributed to cell wall, cell membrane and cytoplasm fractions, respectively. These results indicated that the cell wall was a major adsorbing fraction of cadmium, lead and copper, and the cell membrane was that of zinc. Total amino acid content per gram of the cell grown in the culture media with heavy metal was higher than that of the cell grown in the culture media without heavy metal, and the content of acidic amino acids, such as aspartic acid(Asp.+Asn.) and glutamic acid(Glu.+Gln.) was higher than that of basic amino acids, such as histidine, lysine and arginine.