• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.670-675
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• 2008

A laccase was isolated from the culture filtrate of the basidiomycete Fomitella fraxinea. The enzyme was purified to electrophoretical homogeneity using ammonium sulfate precipitation, anion-exchange chromatography, and gel-filtration chromatography. The enzyme was identified as a monomeric protein with a molecular mass of 47 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel-filtration chromatography, and had an isoelectric point of 3.8. The N-terminal amino acid sequence for the enzyme was ATXSNXKTLAAD, which had a very low similarity to the sequences previously reported for laccases from other basidiomycetes. The optimum pH and temperature for 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) were 3.0 and $70^{\circ}C$, respectively. The enzyme also showed a much higher level of specific activity for ABTS and 2,6-dimethoxyphenol (DMP), where the $K_m$ values of the enzyme for ABTS and 2,6-DMP were 270 and $426{\mu}M$, respectively, and the $V_{max}$ values were 876 and $433.3{\mu}M/min$, respectively. The laccase activity was completely inhibited by L-cysteine, dithiothreitol (DTT), and sodium azide, significantly inhibited by $Ni^+,\;Mn^{2+}$, and $Ba^{2+}$, and slightly stimulated by $K^+$ and $Ca^{2+}$.

• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.648-657
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• 2008

In this study, we used functional genomic strategies, proteomics and quantitative real-time (qRT)-PCR, to advance our understanding of genes associated with fumonisin production in the fungus Fusarium verticillioides. Earlier studies have demonstrated that deletion of the FCC1 gene, which encodes a C-type cyclin, leads to a drastic reduction in fumonisin production and conidiation in the mutant strain (FT536). The premise of our research was that comparative analysis of F. verticillioides wild-type and FT536 proteomes will reveal putative proteins, and ultimately corresponding genes, that are important for fumonisin biosynthesis. We isolated proteins that were significantly upregulated in either the wild type or FT536 via two-dimensional polyacrylamide gel electrophoresis, and subsequently obtained sequences by mass spectrometry. Homologs of identified proteins, e.g., carboxypeptidase, laccase, and nitrogen metabolite repression protein, are known to have functions involved in fungal secondary metabolism and development. We also identified gene sequences corresponding to the selected proteins and investigated their transcriptional profiles via quantitative real-time (qRT)-PCR in order to identify genes that show concomitant expression patterns during fumonisin biosynthesis. These genes can be selected as targets for functional analysis to further verify their roles in $FB_1$ biosynthesis.

• Journal of Ginseng Research
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• v.23 no.1 s.53
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• pp.50-54
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• 1999

In this paper, the saponin enzymatic hydrolysis of ginsenoside Rg3 was studied. The $ginsenoside-{\beta}-glucosidase$ from FFCDL-48 strain mainly hydrolyzed the ginsenoside Rg3 to Rh2, the enzyme from FFCDL-00 strain hydrolyzed Rg3 to the mixture of Rh2 and protopanaxadiol (aglycon). The $ginsenoside-{\beta}-glucosidase$ from FFCDL-48 strain was purified with a column of DEAE-Cellulose to one spot in the SDS polyacrylamide gel electrophoresis. During the purification, the enzyme specific acitvity was increased about 10 times. The purified $ginsenoside-{\beta}-glucosidase$ can hydrolyze the Rg3 to Rh2, but do not hydrolyze the $p-nitrophenyl-{\beta}-glucoside$ which is a substrate of original exocellulase such as ${\beta}-glucosidase$ of cellulose. The molecular weight of $ginsenoside-{\beta}-glucosidase$ was 34,000, the optimal temperature of enzyme reaction was $50^{\circ}C,$ and the optimal pH was 5.0.

• Korean Journal of Clinical Laboratory Science
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• v.37 no.2
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• pp.78-83
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• 2005

The optimum conditions of Cordyceps militaris mycelial growth, purification and characteristics of branched-chain amino acid aminotransferase [BCAT(EC 2.6.1.42)] in this mycelium were studied. Optimum pH, temperature and medium of culture of mycelia were 5.5, $22.5^{\circ}C$ and Hamada medium (HM), respectively. BCAT in homogenate of this mycelia was precipitated by 20-40% saturated solution of ammonium sulfate and then purified by DEAE (diethylaminoethyl)-Sephadex A-50 column chromatography with linear concentration gradient and Sephadex G-200 gel filtration. A single band of purified enzyme was detected on SDS-PAGE (sodium dodecylsulfate-polyacrylamide gel electrophoresis). Optimum pH and temperature of BCAT were found to be 7.8 and $29^{\circ}C$, respectively. It showed activity toward L-leucine, L-isoleucine and L-valine as a substrate. The Km values of this enzyme for L-leucine were determined to be 5.88 mM for L-leucine.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.44 no.6
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• pp.43-49
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• 2012

In this work, the non-ionic polyethylene oxide(PEO)/phenolic formaldehyde resin(PFR) retention system, which was less affected by furnish charge, was analyzed for possibility and effectiveness when using recycled stock. When the micropolymer was added at the PEO/PFR retention system and the PAM/bentonite retention system, performance of retention and drainage was improved. When the cationic micropolymer was added on the PEO/PFR retention system, the highest retention was achieved. The Polyacrylamide(PAM)/bentonite system induced flocculation of white pitch by electrostatic flocculation. On the other hand, the non-ionic PEO/PFR retention system induced less flocculation of white pitch than the PAM/bentonite retention system.

• Journal of Animal Science and Technology
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• v.62 no.5
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• pp.741-752
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• 2020

Herein, the in vitro protein digestibility of lyophilized Protaetia brevitarsis larvae flour with and without defatting using 70% ethanol was compared with beef loin. Proximate analysis showed that the defatted larvae contained the highest protein content (p < 0.05). The viable counts of total aerobic bacteria, Escherichia coli, and coliform bacteria decreased significantly after defatting the larval samples with 70% ethanol (p < 0.05). Measurement of α-amino group content and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed higher amounts of low molecular weight proteins in the larvae compared to beef loin (p < 0.05). After in vitro digestion, the degree of protein hydrolysis of the digesta was higher for both larvae samples compared to beef loin (p < 0.05). No change was observed in the in vitro larval protein digestibility after defatting. These results highlight the excellent protein digestibility of P. brevitarsis larvae with high protein content. Defatting insect flour with 70% ethanol could enhance microbial safety while maintaining excellent protein digestibility.

• Bulletin of the Korean Chemical Society
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• v.31 no.2
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• pp.406-414
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• 2010

A novel polyacrylamide grafted hydrous ferric oxide adsorbent composite has been synthesized by using glow discharge electrolysis plasma. To optimize the synthesis conditions, the following parameters were examined in detail: applied power, discharge time, post polymerization temperature, post polymerization time, amount of crosslinking agent and hydrous ferric oxide gel added and so on. The adsorbent was characterized by Fourier transform infrared spectroscopy (FT-IR) and X-ray photoelectron spectroscopy (XPS). The removal percentage of the adsorbent in Pb(II) solution was examined and the data obtained showed that the adsorbent composite has a high capacity for lead ion. For the use in wastewater treatment, the thermodynamic and kinetic of Pb(II)-capture were also studied. Results indicated that the adsorption reaction was a spontaneous and an endothermic process, and it seems to be obeyed a pseudo-secondorder rate model. Moreover, the adsorption isotherm of Pb(II)-capture is following the Langmuir and Freundlich isotherm models.

• Bulletin of the Korean Chemical Society
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• v.31 no.2
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• pp.275-280
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• 2010

Actinobacillus actinomycetemcomitans is a gram-negative, nonmotile coccobacillus bacterium that is associated with several human diseases, including endocarditis, meningitis, osteomyelitis, subcutaneous abscesses and periodontal diseases. A full-length Omp1-like protein gene from A. actinomycetemcomitans was cloned into a pQE30 vector and overexpressed in Escherichia coli BL21(DE3) cells. The protein revealed sequence homologies to Seventeen kilodalton proteins (Skp) from Pasteurella multocida and E. coli that have been characterized as periplasmic chaperones. This soluble Omp1-like protein was successfully purified to homogeneity for further folding and functional studies. The purity, identity, and conformation of the protein were determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis, matrix-assisted laser desorption ionization mass spectrometry, circular dichroism, fluorescence spectroscopic, and differential scanning calorimetric studies. We showed that the protein formed an oligomer larger than a tetramer. We found, further, that it is comprised of mostly $\alpha$-helices and boasts high thermal stability.

• KSBB Journal
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• v.29 no.5
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• pp.380-384
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• 2014

Inhibition effects of peptide hydrolysates from Astragalus membranaceus Bunge. on the expression of the matrix metalloproteinases (MMPs) in human dermal fibroblasts were evaluated in vitro. Crude peptides were obtained by the hydrolysis of proteins extracted from A. membranaceus. Peptides were purified partially by the basis on the molecular weight using 40% polyacrylamide gel electrophoresis before treatment with human dermal fibroblasts. Basis on the doseeffect experiments, expressions of MMPs including MMP-1, MMP-3, MMP-8, MMP-13 in human dermal fibroblasts were evaluated. Expressions of MMP-1, MMP-3, MMP-8 and MMP-13 were reduced in 43%, 5%, 22% and 57% respectively. The mass spectrometric analysis of partially purified peptides from A. membranaceus, which strongly inhibit expressions of MMPs, indicated that the peptides were composed of molecules below 1500 Da.

• Advances in concrete construction
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• v.9 no.1
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• pp.33-41
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• 2020

In this study, polyethylene glycol (PEG) and polyacrylamide (PAM) have been used as self-curing agents to produce self-curing concrete (SC). Compressive strength, ultrasonic pulse velocity (UPV), bulk electrical resistivity, chloride ion penetrability, water permeability, and main microstructural characteristics were examined under different curing regimes, and compared to those of the control concrete mixture with no self-curing agents. One batch of a control mixture and one batch of a SC mixture were air-cured in the lab to act as non-water-cured samples. The water curing regimes for the control mixture included continuous water curing for 3, 7, and 28 days and periodical moist curing using wetted burlap for 3 and 7 days. Curing regimes for the SC mixtures included 3 days of water curing and periodical moist curing for 3 and 7 days. SC mixtures showed better microstructure development and durability performance than those of the air-cured control mixture. A short water curing period of 3 days significantly improved the performance of the SC mixtures similar to that of the control mixture that was water cured for 28 days. SC concrete represents a step towards sustainable construction due to its lower water demand needed for curing and hence can preserve the limited water resources in many parts of the world.