• Journal of Microbiology and Biotechnology
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• v.16 no.4
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• pp.647-650
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• 2006

D-Psicose 3-epimerase (DPE) catalyzes the interconversion of D-fructose to D-psicose by epimerizing the carbon-3 position. The DPE from Agrobacterium tumefaciens was cloned and expressed in Escherichia coli. The expressed enzyme was purified by affinity chromatography on an IMAC, gel filtration chromatography on a Sephacryl S-300 HR, and anion-exchange chromatography on a RESOURCE Q. The molecular mass of the purified enzyme was estimated to be about 135 kDa by Superdex 200 gel filtration chromatography, corresponding to a homotetramer. The enzyme produced crystals suitable for X-ray diffraction to a $2.0{\AA}$ resolution at 100 K. The crystals were found to belong to the orthorhombic space group $P2_12_12_1$, with unit-cell parameters a=102.4, b=113.0, and $c=131.8{\AA}$. In addition, the calculated packing parameter $(V_m)$ was $2.79{\AA}^3/Da$, the solvent content was 55.92%, and an asymmetric unit consisted of four monomers.

• Journal of Microbiology
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• v.43 no.4
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• pp.370-374
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• 2005

Bacteriocin ST311LD is approximately 2.3 kDa in size. Low levels of bacteriocin activity were recorded in BHI and M17 broth (800 AU/ml) and in $10\%$ (w/v) soy milk (3,200 AU/ml). No bacteriocin pro-duction was recorded in $10\%$ (w/v) molasses, despite good growth. Optimal levels (12,800 AU/ml) were detected in MRS broth which had been supplemented with tryptone (20.0 g/l), saccharose (5.0 or 10.0 g/l) or vitamin C (1 ppm). Increased potassium levels did not result in higher levels of activity, and glycerol (1.0 g/l) inhibited the production of bacteriocin ST311LD.

• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.914-920
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• 2011

Vibrio cholerae utilizes mannitol through an operon of the phosphoenolpyruvate-dependent phosphotransferase (PTS) type. A gene, mtlD, encoding mannitol-1-phosphate dehydrogenase was identified within the 3.9 kb mannitol operon of V. cholerae. The mtlD gene was cloned from V. cholerae O395, and the recombinant enzyme was functionally expressed in E. coli as a $6{\times}$His-tagged protein and purified to homogeneity. The recombinant protein is a monomer with a molecular mass of 42.35 kDa. The purified recombinant MtlD reduced fructose 6-phosphate (F6P) using NADH as a cofactor with a $K_m$ of $1.54{\pm}0.1$ mM and $V_{max}$ of $320.8{\pm}7.81\;{\mu}mol$/min/mg protein. The pH and temperature optima for F6P reduction were determined to be 7.5 and $37^{\circ}C$, respectively. Using quantitative real-time PCR analysis, mtlD was found to be constitutively expressed in V. cholerae, but the expression was up-regulated when grown in the presence of mannitol. The MtlD expression levels were not significantly different between V. cholerae O1 and non-O1 strains.

• Journal of the Korean Wood Science and Technology
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• v.35 no.6
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• pp.159-165
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• 2007

An extracellular xylanase from the brown-rot fungus Fomitopsis palustris grown on rice straw culture was purified to a single protein band. On SDS-PAGE, the molecular mass of purified enzyme was estimated to be about 43 kDa. The amino acid sequence of the proteolytic fragments showed high homology with fungal glycoside hydrolase family 10 xylanases. The $K_m$, $K_{cat}$ and $V_{max}$ for birch xylan were $31mg/m{\ell}$, $2.3{\times}10^4/min$ and 252.3 U/mg, respectively. The optimal activity of the purified xylanase from F palustris was observed at pH 4.0~5.0 and $70^{\circ}C$.

• BMB Reports
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• v.49 no.11
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• pp.612-616
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• 2016

CD44 pre-mRNA includes 20 exons, of which exons 1-5 ($C_1-C_5$) and exons 16-20 ($C_6-C_{10}$) are constant exons, whereas exons 6-15 ($V_1-V_{10}$) are variant exons. $V_6$-exon-containing isoforms have been known to be implicated in tumor cell invasion and metastasis. In the present study, we performed a SR protein screen for CD44 $V_6$ splicing using overexpression and lentivirus-mediated shRNA treatment. Using a CD44 $V_6$ minigene, we demonstrate that increased SRSF3 and SRSF4 expression do not affect $V_6$ splicing, but increased expression of SRSF1, SRSF6 and SRSF9 significantly inhibit $V_6$ splicing. In addition, using a constitutive exon-specific primer set, we could not detect alterations of CD44 splicing after SR protein-targeting shRNA treatment. However, using a $V_6$ specific primer, we identified that reduced SRSF2 expression significantly reduced the $V_6$ isoform, but increased $V_{6-10}$ and $V_{6,8-10}$ isoforms. Our results indicate that SR proteins are important regulatory proteins for CD44 $V_6$ splicing.

• Food Science and Biotechnology
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• v.15 no.6
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• pp.866-870
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• 2006

The maintenance of physiological activity during dilution is very critical for the accurate enumeration of Vibrio spp. in marine samples. We investigated the effect of various diluents on the recovery of Vibrio parahaemolyticus using the direct plate counting and most probable number (MPN) methods. The effects of NaCl (0.85 and 3%) and pH (from 6.6 to 7.4) in diluents based on distilled water or phosphate buffered saline (PBS) were evaluated with three V. parahaemolyticus strains. PBS-3% NaCl (pH 6.6), as opposed to PBS, was the most effective diluent at maintaining viable cell numbers up to 2 log CFU/g during dilution for direct plate counting using on thiosulfate citrate bile salts sucrose (TCBS) selective agar, as well as minimizing the difference in cell numbers between TCBS and non-selective nutrient agar. It also increased counts of V parahaemolyticus inoculated into oysters relative to PBS (p<0.01), suggesting that PBS-3% NaCl (PH 6.6) can reduce the problem of underestimating V. parhaemolyticus counts using PBS alone.

• Korean Chemical Engineering Research
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• v.46 no.3
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• pp.626-632
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• 2008

The purpose of this study was to enhance gas exchange by producing hemosome, a hemoglobin microencapsulated with phospholipid of egg, and perfluorocarbone(PFC) emulsion solution. In the experiment, stable emulsion solution with 437 nm of mean particle size could be produced by Flusol-DA sonication, and shortening of emulsion time could be attained with higher stability as well. $0.8{\mu}m$ sized hemosome could be produced by microencapsulation of hemoglobin with phospholipid extracted from egg yolk. The pattern of oxygen saturation curve of hemosome was S shape, which is similar to that found in normal blood, and $P_{50}$ was measured to be 24 mmHg. The oxygen saturation in the mixed solution of hemosome and blood in 1:4(V/V%) ratio was similar to that of normal blood, and the same result was found in the mixed solution of PFC emulsion and blood in 1:4(V/V%) ratio.

• Korean Journal of Veterinary Research
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• v.38 no.3
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• pp.614-628
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• 1998

The effects of electroacupuncture(EA) on blood concentration of endocrine substances were investigated in 6 horses. Three acupuncture points ; Guan Yuan Shu(BL-26), Wei Shu(BL-21) and Da Chang Shu(BL-25) were stimulated for 20 minutes by EA at separate occasions under varying condition ; 2V-1Hz, 2V-5Hz, 2V-30Hz, 4V-1Hz, 4V-5Hz and 4V-30Hz. Plasma levels of adrenocorticotropic hormone(ACTH), ${\beta}$-endorphin, epinephrine, norepinephrine and serum levels of gastrin were analysed. Blood samplings were carried out before, 0, 20 and 40 minutes after the EA stimulation. The serum gastrin levels were increased by 2V-5Hz stimulation on the Wei Shu. Plasma ACTH levels were decreased by 2V-1Hz stimulation on the Wei Shu, but largely increased by 4V-30Hz stimulation on the Guan Yuan Shu. Plasma ${\beta}$-endorphin levels were slightly increased or decreased by 2V-1Hz stimulation, but largely increased by 4V-30Hz stimulation on the Guan Yuan Shu. Plasma levels of epinephrine and norepinephrine were not so much changed by 2V-1Hz or 5Hz stimulation, but tended to increase by 4V-30Hz stimulation on Guan Yuan Shu. These results suggest that the low voltage-low frequence EA stimulation increased blood concentration of gastrin, but decreased ACTH, ${\beta}$-endorphin, epinephrine and norepinephrine, whereas high voltage-high frequence EA stimulation induced opposite results. Accordingly, there appears to be a close relationship between the changes of gastrointestinal motility and the changes of blood concentration of endocrine substances by EA stimulation.

• Korean Journal of Microbiology
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• v.51 no.2
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• pp.115-125
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• 2015

A monoclonal antibody AP64 IgM binds to human acute nonlymphocytic leukemia (ANLL) cell line HL60 and also cross-reacts with the homologous antigen in a rat ANLL cell. This antibody mediated by complement, has leukemia a suppression effect. In this study, we generated a recombinant single-chain variable domain fragment (ScFv) which were derived from $V_H$ and $V_L$ cDNA of AP64 IgM-secreting hybridoma by RT-PCR. The two variable regions were joined with a single 15 amino acid linker $(G_4S)_3$. This recombinant ScFv was expressed as a single polypeptide chain from Escherichia coli BMH 71-18. The recombinant ScFv was purified by applying the periplasmic extract to $Ni^+$-NTA-agarose affinity column and detected with westernblot. The purified recombinant ScFv recognized a surface antigen (about 30 kDa) of HL60 cell line which is the same antigen detected by parental AP64 IgM. But the affinity of ScFv for a surface antigen of HL60 was lower than that of the parental AP64 IgM, which needs to be further improved. Overall, the recombinant ScFv specific to HL60 might be a useful bioreagent for either diagnostic or therapeutic purposes.

• Journal of the Korean Applied Science and Technology
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• v.34 no.2
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• pp.338-348
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• 2017

Protein hydrolysates derived from plants and animals having antioxidant, suppression of hypertension, immunodulatory, alleviation of pain, and antimicrobial activity has been known as playing important role like hormone. This study was performed to optimize the hydrolysis of protein of sea of cucumber by a flavourzyme. The ranges of processes were the reaction temperature of 40 to $60^{\circ}C$, pH 6 to 8, and enzyme concentration 0.5 to 1.5%(w/v). As a result, the optimization of process was determined at temperature of $48-50^{\circ}C$, pH of 7.0-7.2, and enzyme concentration of 1.0-1.1%(w/v), and degree of hydrolysis was 43-45 at above conditions. The molecular weight of hydrolysate was distributed to 500-3,500 Da and showed typical peptides. Inhibition concentration ($IC_{50}$) of peptides of DPPH radical scavenging activity, Superoxide anion radical scavenging activity, Hydroxy radical scavenging activity, $Fe^{2+}$ cheating activity was 1.25, 3.40, 10.3, and 22.11 mg/mL, respectively. Therefore, we expect that those products are useful as functional food ingredients.