• KSBB Journal
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• v.8 no.2
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• pp.126-132
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• 1993

The objective of this study was to develop an artificial dentin for easy handle and accurate observation of the mechanism on dental caries and to screen biologically active materials from the extracts of traditional plants and fruits for prevention of early dental cares. In order to produce disc PAHA (artificial dentin), the powdered hydroxylapatite was immobilized in a 20% polyacrylamide gel. The characteristics of disc PAHA was very similar to the surface, figure and lattice of human enamel. After decalcification in 0.1M citric acid based on observation with SEM. The critical point of decalcification of disc PAHA by acids was found to be pH 5.0-5.5, which was hi agreement with human enamel. The degree of decalcification from disc PAHA in 0.1M citric acid solution was sixfold higher than that of human enamel. This result suggested that disc PAHA would be useful as a substitute of human enamel for in vitro experiment. The extracts of garlic and Flower Apple A, B seemed to inhibit growth of S. mutans. Especially, when the 300$\mu\ell$ of its extracts added to the medium to incubate S. mutans, F. apple B showed strongly an inhibitory effect in both the growth of S. mutans and the synthesis of insoluble glucan.

• Journal of Microbiology and Biotechnology
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• v.1 no.2
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• pp.102-110
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• 1991

Alkalophilic Bacillus sp. YJ-451, which was isolated from soil at several area in Korea, produced a novel type of bacteriolytic enzyme (cell wall peptidoglycan hydrolase) extracellulary. The cell wall hydrolytic activity was identified as a clear zone on sodium dodecyl sulfate polyacrylamide gel electrophoresis containing 0.2% (w/v) cell wall of Bacillus sp. as substrate. This enzyme was successively purified 66 fold with 3.2% yield in culture broth by ammonium sulfate precipitation, CM-cellulose column chromatography, and gel filtration, followed by hydroxylapatite column chromatography. The molecular weight of the purified enzyme was estimated to be 27,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum pH and temperature for the activity of the enzyme were pH 10.0 and $50^{\circ}C$, respectively. The enzyme was stable between pH 5.0 and 10.0 and up to $40^{\circ}C$. Among the microorganisms used in this experiment the enzyme was active against most of gram negative strains and the genus Bacillus such as B. megaterium, B. licheniformis, B. circulans, B. pumilus, B. macerans, B. polymyxa. The release of dinitrophenylglutamic acid but not reducing group from cell wall peptidoglycan digested by the enzyme suggested that the enzyme is a kind of peptidase which hydrolyzes the peptide bond at the amino group of D-glutamic acid in the peptidoglycan.

• Journal of Applied Biological Chemistry
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• v.42 no.3
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• pp.107-112
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• 1999

A carboxymethyl-cellulase (CMCase) was purified from the culture supernatant of Bacillus sp. KD1014 by ultrafiltration, ammonium sulfate precipitation, and a series of chromatography on QAE-Sephadex A-50, hydroxylapatite and Sephadex G-75. The purified CMCase was a single protein of 32 kDa, showed an optimum activity at $60^{\circ}C$ and pH 6.0, and had a half-life of 23 min at $70^{\circ}C$. The enzyme activity was not influenced by metal ions such as $Mg^{2+},\;Fe^{3+},\;K^+,\;Zn^{2+}$, and $Cu^{2+}$ at a concentration of 1.0 mM, partially inhibited by $Mn^{2+}$ and $Ag^+$, and significantly inhibited by pentachlorophenol (PCP). The purified enzyme showed a 3.9-times higher activity on lichenan than on CMC, but hardly cleaved xylan, starch, avicel, laminarin, filter paper and levan. The results of activity staining of the purified enzyme separated by native and denaturing gel electrophoresis suggested that the CMCase might exist in dimeric, oligomeric or aggregated form as well as in monomeric form. The enzymatic cleavage products from cellotetraose indicated that the CMCase possessed transglycosylation activity.

• BMB Reports
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• v.35 no.2
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• pp.228-235
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• 2002

Methionine aminopeptidase (MetAP) catalyzes the removal of an amino-terminal methionine from a newly synthesized polypeptide. The enzyme was purified to homogeneity from Bacillus stearothermophilus (KCTC 1752) by a procedure that involves heat precipitation and four sequential chromatographs (including DEAE-Sepharose ion exchange, hydroxylapatite, Ultrogel AcA 54 gel filtration, and Reactive red 120 dye affinity chromatography). The apparent molecular masses of the enzyme were 81,300 Da and 41,000 Da, as determined by gel filtration chromatography and sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), respectively. This indicates that the enzyme is comprised of two identical subunits. The MetAP specifically hydrolyzed the N-terminal residue of Met-Ala-Ser that was used as a substrate, and exhibited a strong preference for Met-Ala-Ser over Leu-Gly-Gly, Leu-Ser-Phe, and Leu-Leu-Tyr. The enzyme has an optimal pH at 8.0, an optimal temperature at $80^{\circ}C$, and pI at 4.1. The enzyme was heat-stable, as its activity remained unaltered when incubated at $80^{\circ}C$ for 45 min. The Km and Vmax values of the enzyme were 3.0mM and 1.7 mmol/min/mg, respectively. The B. stearothernmophilus MetAP was completely inactivated by EDTA and required $Co^{2+}$ ion(s) for activation, suggesting the metal dependence of this enzyme.

• BMB Reports
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• v.32 no.3
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• pp.239-246
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• 1999

A tripeptidase (PepT) was purified to homogeneity from Bacillus subtilis through four sequential chromatographies including DEAE-Sepharose ion exchange, hydroxylapatite, mono-Q FPLC ion exchange, and Superose-12 FPLC gel filtration. The apparent molecular mass of the enzyme was 49,200 Da and 51,400 Da as determined by sodium dodecylsulfatepolyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography, respectively, and the enzyme exists in a monomeric form. The physicochemical properties of the enzyme were as follows: optimum pH at 7.5, optimum temperature at $60^{\circ}C$, and pI at 4.9. The $K_m$ and $V_{max}$ values of the enzyme were 4.3 mM and 2.5 mmol/min/mg, respectively, with MetAla-Ser as substrate. The B. subtilis PepT requires $Co^{2+}$ ion(s) for activation, while it is inactivated by EOTA and 1,10-phenanthroline, suggesting that it is a metalloprotein. The enzyme was not inhibited by any of serine protease, aspartic protease, or leucine aminopeptidase inhibitors. The enzyme showed comparable activities towards four different substrates including Met-Ala-Ser, Leu-Gly-Gly, Leu-Ser-Phe, and Leu-Leu-Tyr. The amino terminal sequence of PepT determined by Edman degradation was found to be MKEEIIERFTTYVXV and turned out to be identical to that of PepT deduced from a cloned B. subtilis pepT.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.37 no.5
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• pp.380-385
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• 2011

Introduction: Hydroxyapatite ($Ca_{10}(PO_4)_6(OH)_2$, HA) is the main inorganic phase of human hard tissue that is used widely as the repair material for bones. When HA is applied to a bony defect, however, it can be encapsulated with fibrous tissue and float in the implanted area due to a lack of consolidation. Bioceramics as allogenic graft materials are added to HA to improve the rate and bone healing capacity. Fluoridated hydroxyapatite ($Ca_{10}(PO_4)_6(OH,F)_2$, FHA), where F- partially replaces the OH- in hydroxyapatite, is considered a good alternative material for bone repair owing to its solubility and biocompatibility. Materials and Methods: This study was designed to determine the bone healing capacity of FHA newly produced as a nanoscale fiber in the laboratory. HA and FHA with bioglass was implanted in a rabbit cranium defect and the specimen was analysed histologically. Results: 1. At 4 weeks, fibrous connective tissue and little bone formation was observed around the materials of the experimental group I implanted HA and bioglass. Newly formed bone was observed around the materials in the experimental group II implanted FHA and bioglass. 2. At 8 weeks, the amount of newly formed and matured bone was higher in experimental group II than in experimental group I and the control group. Conclusion: These results suggest that FHA and bioglass is a relatively favorable bone substitute with biocompatibility and better bone healing capacity than pure HA and bioglass.

• Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
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• v.19 no.2
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• pp.117-122
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• 2008

Background and Objectives: $Radiesse^{(R)}$ is a gell-formed material of calcium hydroxylapatite (CaHA) and carboxymethylcellulose (CMC) used for vocal fold injections. The authors aimed to study injection laryngoplasty with $Radiesse^{(R)}$, and determine the efficacy of $Radiesse^{(R)}$ for unilateral vocal cord palsy using objective and subjective measures. Materials and Method: Nine patients with unilateral vocal cord palsy received injection laryngoplasty with $Radiesse^{(R)}$ under general anesthesia from Jul. 2007 to Jan. 2008. $Radiesse^{(R)}$ was injected with 25gauze long needle perorally or percutaneously. The Acoustic, aerodynamic, stroboscopic analysis and pre-injection/post-injection perceptual assessment were evaluated in all patients. Results: Postoperative jitter and shimmer scores tended to diminish compared to preoperative scores, and maximum phonation time increased with statistical significantly. Stroboscopic findings demonstrated improvement postoperatively. The degree of hoarseness, which is a part of perceptual assessment, decreased after the procedure. Conclusion: $Radiesse^{(R)}$ may be an altemative material for injection laryngoplasty. We suggest long-term follow up with more cases.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.576-583
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• 2004

GERI-155 is a macrolide antibiotic containing two deoxyhexose molecules and shows antimicrobial activities against Gram-positive bacteria. Deoxysugar biosynthetic gene cluster of GERI-155 from Streptomyces sp. GERI-l55 genome was cloned. Four orfs were identified and a putative orf presumed to be the dTDP g]ucose-4,6-dehydratase gene was designated as gerE. GerE was expressed in E. coli by using a recombinant expression vector pHJ1. The expressed protein was purified from E. coli cell lysate by using ammonium sulfate fractionation, and DEAE-sepharose CL-6B and hydroxylapatite column chromatography. The molecular mass of the expressed protein correlated with the predicted mass that was deduced from the cloned gene sequence data. The recombinant protein was a homodimer with a subunit relative molecular weight of 39,000 Dalton. It was found to have dTDP-glucose 4,6-dehydratase activity and also found to be highly specific for dTDP-glucose as a substrate. The values of $K_{m} and V_{max}$ for dTDP-g]ucose were $32\mu$M and 335 nmol $min^{-1}$ (mg protein)^{-1}$, respectively. dTTP and dTDP were strong inhibitors of the protein. $NAD^+$, the coenzyme for dTDP-glucose 4,6-dehydratase, was tightly bound to the expressed protein.

• Biomedical Science Letters
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• v.2 no.1
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• pp.41-48
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• 1996

The C-reactive protein(CRP) from ascitic and pleural fluid was purified using calcium dependent affinity chromatography of CNBr activated Sepharose-4B covalently coupled to p-diazonium phenylphosphorylcholine(DPPC) and hydroxylapitite chromatography. Polyclonal antibody was prepared from rabbit by immunizing the purified CRP. Specific immunoglobulin G was isolated using affinity chromatography and coupled to microparticles. A sensitive microparticle-based immunoassay was developed to measure CRP within 3 mins. The detection range was between 0.5mg/dl) and 20mg/dl in serum, showing strong response in the range of 0.7~2.9 mg/dl, weak response in 5.0~13.2 mg/dl and zone phenomenon over 28mg/dl. The average value of CRP in 74 samples was 3.8mg/dl and most of the values were lower than 10mg/dl .The CRP values of serum samples were determined by our microparticle-based immunoassay, and were compared with those obtained using the other commercial products(B Co., France and I Co., Japan). Good correlations were shown between the values obtained by our developed microparticle-based immunoassay system and those by other commercial products. All performance characteristics evaluated make our developed microparticles-based immunoassay suitable for a simple, rapid, and reliable screening of CRP in serum.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.27 no.1
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• pp.15-24
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• 2001

The purpose of this study is to evaluate the tissue response in applying of various bone substitutes included toothash-plaster mixture, resorbable hydroxylapatite (HA) and demineralized freeze-dried bone and to show the clinical usefulness of toothash-plaster mixture for the repair of craniomaxillofacial bone defect. For this experiment, 100 Sprague-Dawley rats weighing 200gm or more were used. There were four experimental groups: group I, toothash-plaster mixture; group II, demineralized freeze-dried bone; group III, resorbable HA; and group IV, control group. A full thickness, round bone defect measuring 10mm in diameter was created in the midcranium, and the substitutes cited above were embedded in the experimental rats based on their group assignment. Blood clot was filled in the rats assigned to the control group. Experimental rats were sacrificed on the 1st, 3rd, 5th, 8th, 12th and 24th week after implantation and stained with the hematoxylineosin, Masson's Trichrome, using Van Gieson's stain method, and were examined under light microscope. The results were as follows: 1. In all the groups, prominent inflammatory reaction and the infiltration of multinucleated giant cells were noted during the early stage. Gradual healing decreased this reaction. 2. Among the rats in the experimental group II, which were given demineralized freeze-dried bone implants, active formation of new bone traveculae manifested. Chondroid tissues appeared, and it was suggested that the defect was filled with newly formed bone by virtue of osteoinductive activity. On the 12th week after the experiments, most of the defect was filled with newly formed bone trabeculae. 3. In experimental groups I and III, it was noted that HA manifested a healing process similar to that characterized by the toothash-plaster mixture, but inflammatory reaction was more prominent in experimental group I. Active osteoblasts were observed along the periphery of osteoid tissues, while newly formed bone trabeculae appeared adjacent to the implanted materials three weeks later. Formation increased to the extent that newly formed bone trabeculae fused directly with the host bone. Increase in new bone ingrowth into the filling materials was revealed by both experimental groups. 4. In the control group, new bone formation adjacent to the host bone was observed, but most of the defect was filled with mature connective tissue 24 weeks after the experiments.