• International Journal of Industrial Entomology and Biomaterials
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• v.39 no.2
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• pp.67-73
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• 2019

The genomic structure of the Cu/Zn superoxide dismutase (SOD1) gene from the entomopathogenic fungus, Cordyceps pruinosa was characterized. The SOD1 gene of C. pruinosa spans 947 nucleotides and consisted of four exons encoding for 154 amino acids and three introns. Four exons of the SOD1 gene are composed of 13, 331, 97 and 20 nucleotides respectively. Homology search of amino acid sequences of the SOD1 gene of C. pruinosa with another 13 fungi species showed higher sequence similarity of 69% ~ 95% and had the most highest sequence identity of 95% with Beauveria bassiana and Cordyceps militaris, which can easely infect domesticated Bombyx mori and another wild lepidopteran species in artificial or natual manner of infection. This SOD1 gene sequence showed copper, zinc and beta-barrel fold sites. Homology search showed that the Cu/Zn SOD1 gene from the entomopathogenic fungus, C. pruinosa is an orthologous gene homolog present in different species of organism whose ancestor predates the split between the relating species. In addition, C. pruinosa SOD1 gene is placed together within the ascomycetes group of fungal clade. From these results it is concluded that C. pruinosa SOD1 gene is orthologous gene having the same or very similar functions with a common evolutionary ancestor.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.6
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• pp.574-579
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• 2009

Acid- and pepsin-solubilized collagens were extracted from the skin of shark (Isurus oxyrinchus) and their physicochemical properties were characterized by amino acid analysis, SDS-PAGE, the composition of collagen types, solubility and denaturation temperature. Acid - and pepsin-solubilized collagens from shark skin had an imino acid of 188.8 and 186.2 residues/1,000 amino acids, respectively. SDS-PAGE showed two different${\alpha}$ chains ($\alpha1$ and $\alpha2$) and $\beta$-component. The component ratio of type I and V was 10:1, and the type III was not found. Solubility of acid-soluble collagen was low in the range of pH 6.0 to pH 11.0. On the other hand, pepsin-solubilized collagen showed a low solubility in the range of pH 7.0-9.0. Temperature for denaturation of acid- and pepsin-solubilized collagens were $25^{\circ}C$ and $27^{\circ}C$, respectively.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.243-246
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• 2005

A novel esterase showing high enantioselectivity to (S)-ketoprofen ethyl ester was selected from fosmid environmental DNA library which is provided by Microbial Genomic & Applications Center. As a result of Blast search, the gene wasn't registerated in Gene Bank yet. And as we know, conserved domain region of esterase , G-X-S-X-G, wasn't discovered.$^{4)}$ And it is similar to Beta-lactamase. The DNA sequence of cloned esterase include an open reading frame consisting of 1170 bp, designated as EST-Y29, encoding a protein of 389 amino acids with a molecular mass of about 42.8 kDa. And amino acid sequence analysis revealed only a few identity (28%) to tile known esterases/lipases in the databases containing the conserved sequence motifs of esterases/lipases. when being comparison to other esterase revealed , this enzyme seems to be classified as a new member of esterase family. EST-Y29 was functionally overexpressed in a soluble form in E. coli with maximum conversion yield of (S)-ketoprofen at $65^{\circ}C$. This study demonstrates that functional screening combined with the sequential uses of restriction enzymes to exclude already known enzymes is a useful approach for isolating novel enzyme from a metagenome.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.153-157
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• 2004

Tyrosine phenol-lyase (TPL) is a useful enzyme for the synthesis of pharmaceutical aromatic amino acids. In the current study, sequential DNA shuffling and screening were used to enhance the stability of TPL. Twenty-thousand mutants were screened, and several improved variants were isolated. One variant named A13V, in which the $13^{th}$ amino acid alanine was substituted by valine, exhibited a higher temperature and denaturant stability than the wild-type TPL. The purified mutant TPL, A13V, retained about 60% of its activity at $76^\circ{C}$, whereas the activity of the wild-type TPL decreased to less than 20% at the same temperature. Plus, A13V exhibited about 50% activity with 3 M urea, while the wild-type TPL lost almost all its catalytic activity, indicating an increased denaturant tolerance in the mutant A13V. It is speculated that the substitution of Val for the Ala in the $\beta$-strand of the N-terminal arm was responsible for the heightened stabilization, and that the current results will contribute to further research on the structural stability of TPL.

• Journal of Ginseng Research
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• v.22 no.2
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• pp.73-81
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• 1998

In order to stabilize the price of fresh ginseng by extension of seasonal variation and marketing structure, and to reduce the cost of production by works of intensive and short term, studies were carried out. As fresh ginseng of 4 years old by the MA (modified atmosphere storage) and CA (controlled atmosphere storage) was stored for 12 week at 4$^{\circ}C$, samples were collected after every 1, 2, 3, 4, 6, 8, 12 weeks for processing red ginseng. Chemical characteristics on ginseng steamed red for the quality evaluation are summarized as follows. The composition of free sugar was consisted of fructose, glucose, sucrose, and maltose in fresh ginseng. And small change was revealed in free sugar composition during the storage period. Glucose and sucrose were largely increased in both treatments of MA (modified atmosphere storage) and CA (controlled atmosphere storage) after 4 week, wherein maltose was decreased. Fifteen different amino acids were identified, and total amount of histamine and arginine were over 40%. Phenylalanine was slightly decreased. As the volatile components in control red ginseng, six different ones including triisopropyl benzene were detected. B-1 ($CO_2$ : $O_2$ : N2=6 : 4.90, DF-100) of CA and E-1 (CPP, DF-100) of MA did not shown triisopropyl benzene as the volatile components. And ${\gamma}$-muurolene, guaiaene, and beta-patchoulene were reduced, but the other components showed the trend of increase.

• Journal of Dairy Science and Biotechnology
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• v.29 no.1
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• pp.65-73
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• 2011

Cheese is a nutritious food with various balanced nutrients, such as proteins, peptides, amino acids, fats, fatty acids, vitamins and minerals. Domestic cheese varieties and quality need to be improved to prevent imported cheese. To develop those cheeses, search for previous works and research for new products are needed. In cheese ripening of hard cheese, such as Cheddar or Parmesan cheese, is ripened for 2 to 24 months at 2 to 16$^{\circ}C$ to develop desired cheese flavor and body characteristics. Long time with low temperature to ripen the cheese requires high expenses. So accelerated cheese ripening is a good potential for saving in industry. Methods for acceleration of cheese ripening are temperature control, addition of bacteria or enzymes. To develop the functionality of cheese, addition of microencapsulated various probiotics and nutrients, such as iron, removal of cholesterol by crosslinked ${\beta}$-cyclodextrin, lowering blood cholesterol and serum glucose by nanopowdered functional materials et al. are necessary. Therefore, this review focused on the functionality of cheese, such as the acceleration of cheese ripening, microencapsulated probiotics and iron, and cholesterol removal.

• Journal of the Korean Applied Science and Technology
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• v.3 no.2
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• pp.29-34
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• 1986

The sulfuric acid sulfonation of 2-aminonaphthalene was preformed at $30^{\circ}C{\sim}200^{\circ}C$ for $1{\sim}15$ hours, using $4{\sim}10$ parts of 90% sulfuric acid${\sim}$20% oleum. This reaction was influenced by many factory such as reaction temperature, reaction time, strength of sulfuric acid and amount of sulfuric acid and among these factors, the effect of the reaction temperature was considerable. The main products of this reaction were Dahl's acid (2-aminonaphthalene-5-sulfonic acid, Broenner's-acid (2-aminonaphthalene-6-sulfonic acid), amnio-F-acid(2-aminonapthalene-7-sulfonic acid), and Baden acid (2-aminonaphthalene-7-sulfonic acid) and another isomers such as Tobia's acid (2-aminonaphthalene-1-sulfonic acid) and 2-aminonaphthalene-4-sulfonic acid were not formed. Of these isomeric acids, those containing the sulfonic acid group in an ${\alpha}-position$, namely, Dahl's acid and Baden acid, are simultaneously formed by sulfonation at low-temperature ($30{\sim}100^{\circ}C$), whilst those containing the sulfonic acid group in a ${\beta}$-position, namely, Broenner's acid and amino-F-acid, are formed simultaneously by sulfonation at high temperatures($150^{\circ}{\sim}200^{\circ}$).

• Journal of Microbiology and Biotechnology
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• v.24 no.9
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• pp.1196-1206
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• 2014

A metagenomic fosmid library was constructed using genomic DNA isolated from the gut microflora of Hermetia illucens, a black soldier fly. A cellulase-positive clone, with the CS10 gene, was identified by extensive Congo-red overlay screenings for cellulase activity from the fosmid library of 92,000 clones. The CS10 gene was composed of a 996 bp DNA sequence encoding the mature protein of 331 amino acids. The deduced amino acids of CS10 showed 72% sequence identity with the glycosyl hydrolase family 5 gene of Dysgonomonas mossii, displaying no significant sequence homology to already known cellulases. The purified CS10 protein presented a single band of cellulase activity with a molecular mass of approximately 40 kDa on the SDS-PAGE gel and zymogram. The purified CS10 protein exhibited optimal activity at $50^{\circ}C$ and pH 7.0, and the thermostability and pH stability of CS10 were preserved at the ranges of $20{\sim}50^{\circ}C$ and pH 4.0~10.0. CS10 exhibited little loss of cellulase activity against various chemical reagents such as 10% polar organic solvents, 1% non-ionic detergents, and 0.5 M denaturing agents. Moreover, the substrate specificity and the product patterns by thin-layer chromatography suggested that CS10 is an endo-${\beta}$-1,4-glucanase. From these biochemical properties of CS10, it is expected that the enzyme has the potential for application in industrial processes.

• Bulletin of the Korean Chemical Society
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• v.32 no.10
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• pp.3675-3681
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• 2011

Fusarium oxysporum, an important pathogen that mainly causes vascular or fusarium wilt disease which leads to economic loss. Disruption of gene encoding a heterotrimeric G-protein-${\beta}$-subunit (FGB1), led to decreased intracellular cAMP levels, reduced pathogenicity, colony morphology, and germination. The plant defense protein, Nicotiana alata defensin (NaD1) displays potent antifungal activity against a variety of agronomically important filamentous fungi. In this paper, we performed a molecular modeling and docking studies to find vital amino acids which can interact with various antifungal compounds using Discovery Studio v2.5 and GRAMMX, respectively. The docking results from FGB1-NaD1 and FGB1-antifungal complexes, revealed the vital amino acids such as His64, Trp65, Ser194, Leu195, Gln237, Phe238, Val324 and Asn326, and suggested that the anidulafungin is a the good antifungal compound.The predicted interaction can greatly assist in understanding structural insights for studying the pathogen and host-component interactions.

• Journal of Microbiology and Biotechnology
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• v.6 no.6
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• pp.474-481
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• 1996

Since the commercially available rabbit anti-cyclin D3, generated from c-terminal 16 amino acid residues which are common to human and murine cyclin D3, is highly cross-reactive with many other cellular proteins of mouse, a new rabbit polyclonal anti-cyclin D3 has been raised by using murine cyclin D3 protein expressed at a high level in Escherichia coli as the immunogen. To express murine cyclin D3 protein in E. coli, the cyclin D3 cDNA fragment encoding c-terminal 236 amino acid residues obtained by polymerase chain reaction (PCR) was inserted into the NcoI/BamHI site of protein expression vector, pET 3d. Molecular mass of the cyclin D3 overexpressed in the presence of IPTG (Isopropyl $\beta$-D-thiogalactopyranoside) was approximately 26 kDa as calculated from the reading frame on the DNA sequence, and the protein was insoluble and mainly localized in the inclusion bodies that could be easily purified from the other cellular soluble proteins. When renaturation was performed following denaturation of the insoluble cyclin D3 protein in the inclusion bodies using guanidine hydrochloride, 4.4 mg of soluble form of cyclin D3 protein was produced from the transformant cultured in 100ml of LB media under the optimum conditions. Four-hundred micrograms of the soluble form of cyclin D3 protein was used for each immunization of a rabbit. When the antiserum obtained 2 weeks after tertiary immunization was applied to Western blot analysis, it was able to detect 33 kDa cyclin D3 protein in both murine lymphoma cell line BW5147.G.1.4 and human Jurkat T cells at 3,000-fold dilution with higher specificity to murine cyclin D3, demonstrating that the new rabbit polyclonal anti-murine cyclin D3 generated against c-terminal 236 amino acid residues more specifically recognizes murine cyclin D3 protein than does the commercially available rabbit polyclonal antibody raised against c-terminal 16 amino acids residues.