• Journal of the Korean Society of Food Culture
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• v.5 no.2
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• pp.269-274
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• 1990

This study was conducted to determine invertase activity in persimmon during the drying process and characterize the purified enzyme. As drying proceeded, invertase activity increased until 10 days and decreased gradually afterwards. Invertase in persimmon fruit was extracted with 250 mM potassium phosphate sulfate buffer at pH 7.4. The enzyme was purified by means of ammonium sulfate fractionation, column chromatography on DEAE-cellulose and gel filtration on Sephadex G-200 column. The optimal temperature of enzyme was $40^{\circ}C$ and optimal pH was 5.0 and 6.0 for sucrose and raffinose, respectively. The enzyme was stable up to $50^{\circ}C$ and pH 3-6. The Km value of the enzyme, with sucrose as a substrate, was 2.5mM. Electrophoretic pattern of purified enzyme solution showed a single band.

• Journal of Microbiology
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• v.41 no.3
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• pp.212-218
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• 2003

Citrus phytoene synthase (CitPsy) and ${\beta}$-carotene hydroxylase (CitChx), which are involved in caroteinoid biosynthesis, are distantly related to the corresponding bacterial enzymes from the point of view of amino acid sequence similarity. We investigated these enzyme activities using Pantoea ananatis carotenoid biosynthetic genes and Escherichia coli as a host cell. The genes were cloned into two vector systems controlled by the T7 promoter. SDS-polyacrylamide gel electrophoresis showed that CitPsy and CitChx proteins are normally expressed in E. coli in both soluble and insoluble forms. In vivo complementation using the Pantoea ananatis enzymes and HPLC analysis showed that ${\beta}$-carotene and zeaxanthin were produced in recombinant E. coli, which indicated that the citrus enzymes were functionally expressed in E. coli and assembled into a functional multi-enzyme complex with Pantoea ananatis enzymes. These observed activities well matched the results of other researchers on tomato phytoene synthase and Arabidopsis and pepper ${\beta}$-carotene hydroxylases. Thus, our results suggest that plant carotenoid biosynthetic enzymes can generally complement the bacterial enzymes and could be a means of carotenoid production by molecular breeding and fermentation in bacterial and plant systems.

• Food Science of Animal Resources
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• v.34 no.5
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• pp.570-575
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• 2014

This study was conducted to compare the effects of two forms of radiation (electron and X-ray; generated by an electron beam accelerator) on the conformation and antigenic properties of hen's egg albumin, ovalbumin (OVA), which was used as a model protein. OVA solutions (2.0 mg/mL) were individually irradiated by electron beam or X-ray at the absorbed doses of 0 (control), 2, 4, 6, 8, and 10 kGy. No differences between the two forms of radiation on the structural properties of OVA were shown by spectrometric and electrophoretic analyses. The turbidity of OVA solution increased and the main OVA bands on polyacrylamide gels disappeared after irradiation, regardless of the radiation source. In competitive indirect enzyme-linked immunosorbent assay, OVA samples irradiated by electron beam or X-ray showed different immunological responses in reactions with monoclonal and polyclonal antibodies (immunoglobulin G) produced against non-irradiated OVA. The results indicate that electron beam irradiation and X-ray irradiation produced different patterns of structural changes to the OVA molecule.

• The Plant Pathology Journal
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• v.22 no.3
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• pp.260-264
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• 2006

Talaromyces flavus mediates the transmission of Apple stem grooving virus (ASGV) to several host plants. The ASGV-F carried by T.flavus was partially purified from the fungus. Based on sequence analysis and homology searches, this is closely related to other ASGV strains isolated from host plants. The partially purified viral coat protein (CP) was separated on a 12% SDS-polyacrylamide gel and analyzed by Western blotting with an ASGV anti-serum. A single band at 28 kDa reacted with the ASGV anti-serum. The deduced amino acid sequence of the ORF-l showed conserved domains, including an NTP-binding helicase motif, GFAGSGKT. The amino acid sequences of the helicase and CP showed strong homology to other ASGV strains (98%). All ASGV isolated from plants and fungi had salt bridges composed of the CP and the GFAGSGKT motif of the helicase, which are commonly conserved in plant viruses. These results suggest that ASGV-F is one of ASGV strains isolated from T.flavus based on sequence similarity as well as the serological analysis of CP.

• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.457-464
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• 2008

An extracellular enzyme (RMEBE) possessing ${\alpha}-(1{\rightarrow}4)-(1{\rightarrow}6)$-transferring activity was purified to homogeneity from Rhodothermus marin us by combination of ammonium sulfate precipitation, Q-Sepharose ion-exchange, and Superdex-200 gel filtration chromatographies, and preparative native polyacrylamide gel electrophoresis. The purified enzyme had an optimum pH of 6.0 and was highly thermostable with a maximal activity at $80^{\circ}C$. Its half-life was determined to be 73.7 and 16.7 min at 80 and $85^{\circ}C$, respectively. The enzyme was also halophilic and highly halotolerant up to about 2M NaCl, with a maximal activity at 0.5M. The substrate specificity of RMEBE suggested that it possesses partial characteristics of both glucan branching enzyme and neopullulanase. RMEBE clearly produced branched glucans from amylose, with partial ${\alpha}-(1{\rightarrow}4)$-hydrolysis of amylose and starch. At the same time, it hydrolyzed pullulan partly to panose, and exhibited ${\alpha}-(1{\rightarrow}4)-(1{\rightarrow}6)$-transferase activity for small maltooligosaccharides, producing disproportionated ${\alpha}-(1{\rightarrow}6)$-branched maltooligosaccharides. The enzyme preferred maltopentaose and maltohexaose to smaller maltooligosaccharides for production of longer branched products. Thus, the results suggest that RMEBE might be applied for production of branched oligosaccharides from small maltodextrins at high temperature or even at high salinity.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.27 no.2
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• pp.63-81
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• 1997

This study was performed in order to identify changes of the plasma membrane proteins in rat submandibular gland tumors induced by 7,12-dimethylbenz[a]anthracene [DMBA] and X-irradiation. Two kinds of tumor associated membrane proteins (protein A and B) were isolated with 3 M KCl extraction from rat submandibular gland tumors induced by DMBA and X-irradiation. To identify their antigenicities, immunoelectrophoresis and double immunodiffusion was carried out with various proteins extracted from liver, heart, skin and pancreas of adult rats and from embryonic liver, heart and skin. The rabbit antisera against the protein A did not cross-react with any of the proteins extracted from the above mentioned tissues, suggesting that protein A might be tumor specific antigen. However, the rabbit antisera against protein B was precipitated with proteins extracted from the liver of adult and embryonic rats. Polyacrylamide gel electrophoresis of these two proteins (A and B) showed that protein A was a dimer with molecular weights of 69,000 and 35,000 dalton, whereas protein B was a monomer with molecular weight of 50,000 dalton.

• Journal of the Korean Chemical Society
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• v.23 no.5
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• pp.320-324
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• 1979

Purple membrane from Halobacterium halobium was incorporated into 7.5% polyacrylamide gels. Absorption and circular dichroic spectra of purple membrane incorporated with gels were obtained at various pH. The spectra of these gels measured at pH 7.0 were essentially identical with those obtained in the aqueous suspension of purple membrane. Acid titration of the gels showed the transition to a form absorbing at 605nm $(bR_{605}^{acid}$) at pH 2.6, and to a second form at 565nm $(bR_{565}^{acid})$ at pH 0.8. Dark-adapted gels showed an isosbestic point for each transition whereas light-adapted gels did not. Visible CD spectra of $bR_{570}^{LA},\;bR_{305}^{acid}\;and\;bR_{565}^{acid}$ all showed the typical bilobed pattern. CD spectra measured at UV wavelength region were also independent of the variation of pH in terms of molar ellipticity and spectral shape. The protonated species $bR_{605}^{acid}$ may be one of the intermediates formed during the normal photochemical cycle of purple membrane. Most probably, the species $bR_{605}^{acid}$ is considered to be $O^{640}$ in the cycle.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.47 no.4
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• pp.115-126
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• 2015

In this study, we investigated influence of ground calcium carbonate (GCC) modified by washless multilayering of polyelectrolytes on paper quality. Three layers of polyelectrolytes (cationic starch/anionic polyacrylamide/cationic starch) were formed on the surface of GCC using laboratory inline washless polyelectrolytes multilayering system, which was called inline LbL GCC. Base papers were prepared with untreated GCC or inline LbL GCC using a laboratory handsheet former. These handsheets were coated with rod coater, and then printed by black ink. Properties of base paper and fold crack of coated paper were evaluated. Base paper with inline LbL GCC showed much higher mechanical strength in terms of tensile index, strain, internal bond strength, and folding endurance. The fold crack of coated paper with inline LbL GCC occurred more frequently compared to coated paper with untreated GCC. This might be due to highly improved internal bond strength of base paper, which resulted in smaller delamination that played a role of stress dissipation. It would be recommended to design a proper coating layer in order to prevent fold crack.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.35 no.6
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• pp.353-359
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• 2013

Purpose: Extensive research is actively ongoing for development of an ideal bone substitute that meets the gold standard. Tooth was selected as a donor site for evaluation of potentials in bone substitutes based on its similar chemical compositions to alveolar bone. Previous studies have evaluated inorganic components of autogenous tooth bone graft material (AutoBT) and osteoconductivity. In continuation from the previous studies, the current study was conducted for analysis of organic components and evaluation of osteoinductivity of AutoBT. Methods: Forty-six extracted teeth were collected from actual patients (Korea Tooth Bank, R&D Institute). Extracted teeth were processed into AutoBT and implanted in dorsal subcutaneous muscular tissues of 15 athymic mice. Biopsy samples were harvested at two, five, and eight weeks. The Bradford assay, sodium dodecyl sulphate polyacrylamide gradient gel, and western blotting were performed for investigation of organic contents of AutoBT. Results: Histology analyses showed signs of new bone formation as early as two weeks. Results of the Bradford assay indicated the existence of noncollagenous proteins (NCP). 0.29% (2.89 mg/g) of proteins were extracted by weight in the root portion of AutoBT; 0.02% (0.029 mg/g) and 1.79% (17.93 mg/g) of proteins were measured by weight in crown and block-form of AutoBT, respectively. However, recombinant human bone morphogenetic protein-2 was not observed in AutoBT. Conclusion: Within the limitation of the current study, AutoBT induced new bone formation by NCP embedded in dentin.

• Journal of Korean Clinical Health Science
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• v.1 no.1
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• pp.83-89
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• 2013

Purpose. Natural herb has been used for traditional medicine for thousands of years. For this reason, alternative medicine methods affect to domestic animal field. The main purpose of this study was to confirm the regulated proteins after feed additive treatment. Methods. We used total thirty male pigs were used for this experiment. E. coli and Salmonella typhimurium, were used for promote enteritis. Animals were divided into a negative control group, positive control group and test group to determine the effect of an additive mixture on the changes in protein expression in serum. The pigs were given a food supplemented with a natural herbal additive containing immunoglobulin yolksac (IgY) at concentrations 1%. On the 1st week and after eight weeks of feeding, the serums were collected from each group. The changes in the serum proteome as a response to the herbal additive were examined using two-dimensional polyacrylamide gel electrophoresis. Results. A total of 14 differentially expressed protein spots were identified by comparing the protein profiles of the control and additive treated porcine serums. Finally, 7 proteins were detected by MALDI-TOF/MS. Moreover, the proteins detected are involved in a cholesterol control factor, inflammation regulator, Conclusion. These results support of the hypothesis that a natural herbal additive containing IgY can affect the immune regulation system and reduce the pathological process by microbial infections.