• Korean Journal of Agricultural Science
• /
• v.15 no.2
• /
• pp.153-163
• /
• 1988

This experiment was carried out to obtain the thermophilic fungus producing amylase and to investigate properties of the amylase. The selected strain, L-11 was obtained from soil in the vicinity of a hot spring and identified as Mocor sp.. And then the conditions for enzyme production in wheat bran cultures and properties of the crude enzyme were investigated. Furthermore, the enzyme was purified and the characteristics of purified enzyme were studied. The results obtained were as follows: 1. On the wheat bran medium added 80-100% water, amylase was effectively produced by the selected strain, L-11 for 48 hrs incubation at $50^{\circ}C$. 2. When the crude enzyme solution of the strain L-11 was passed through DEAE-Sephadex A-50 column chromatography, two peaks having amylase activity were obtained, and one peak was that of the main enzyme (enzyme of B peak). 3. The purified enzyme (enzyme of B peak) was recognized as single protein band on polyacrylamide disc gel electrophoresis. 4. In the hydrolysis reaction of soluble starch by the enzyme of main amylase, oligosaccharides produced at early stage were maltose and maltotriose mainly and procedure of the reaction maltose amount of maltose and glucose was increased. 5. The strain L-11 was recognized as a special strain producing ${\alpha}-amylase$ mainly and scarcely glucoamylase. 6. The optimum pH, optimum temperature, pH stability, and temperature stability of ${\alpha}-amylase$ were pH 4.0, $60-65^{\circ}C$, pH 4.0-9.0, and below$70^{\circ}C$.

• Journal of Korean Society of Forest Science
• /
• v.58 no.1
• /
• pp.1-7
• /
• 1982

Megagametophyte and embryo tissue of Pinus densiflora were subjected to study the inheritance of glutamate-oxalate transaminase(GOT) and leucine aminopeptidase(LAP), and linkage relationship among isozyme loci coding both enzymes by starch gel zone-electrophoresis. Four zones of activity were observed for GOT. No variation was found in the fastest migrating zone (GOT-A). Electrophoretic phenotypes of the other two zones (GOT-B and GOT-C) showed 1:1 segregation ration, suggesting that each zone is controlled by a single locus. Foru and three alleles were identified at both loci respectively. The isozyme pattern of the fourth zone(GOT-D), migrated cathodally, coincided precisely with that of GOT-C. Whether the two zones are controlled by the same locus or by two tightly linked loci remained unknown. In all three variant GOT zones, heterozygoes embryos produced triple band patterns, indicating that GOT isozyme in Pinus densiflora is a dimer. Two zones of activity stained for LAP were found. The segregation of the two zones (LAP-A and LAP-B) suggested that tow loci control each of both isozymes. Two and three alleles were identified at both loci. GOT-B and LAP-B were found to be tightly linked, showing an average recombination frequency of 12.5 percent. Slight deviation from independent assortment was observed between GOT-B and GOT-C, with recombination frequency of 41 percent.

• Korean Journal of Biological Psychiatry
• /
• v.7 no.1
• /
• pp.64-73
• /
• 2000

There are several candidate genes in genetic study of alcoholism. Among them, allelic associations have been reported between MAOA CA repeat polymorphism and alcohol dependence, recently. And also, several studies have been investigated genotype-phenotype relationships between MAOA CA repeat polymorphism and clinical manifestations. The authors tried to identify differences in allelic frequency of MAOA CA repeat polymorphism between alcohol dependence and controls, and in behavioral trait and clinical characteristics according to MAOA CA repeat polymorphism. We also tried to investigate genotype-phenotype relationships between MAOA CA repeat polymorphism and behavioral trait such as aggression. We examined 49 male patients with alcohol dependence(DSM-IV) who had been admitted in Yong-In Mental Hospital from June 1st 1998 to October 31th 1998. We performed semistructured interview for demographic and clinical characteristics. Self-report questionnaire for BDHI(Buss-Durkey Hostility Inventory) was given to all subject at least 4weeks later after admission. Using polymerase chain reaction and polyacrylamide gel electrophoresis, MAOA CA repeat polymorphism were observed in 52 male controls and 49 male patients with alcohol dependence. We devided alcoholic patients into two groups according to allelic length of MAOA CA repeat polymorphism ; alcoholics with short alleles(${\leq}$119bp, N=20) and alcoholics with long alleles(${\geq}$123bp, N=29). T-test, ${\chi}^2$-test and Fisher exact probability test were used for statistical analysis. There were no significant differences in frequency of each allele and short and long alleles of MAOA CA repeat polymorphism between alcoholics and controls. But there were significant differences in clinical symptoms and behavioral trait between alcoholics with short and long alleles. In clinical symptoms, alcoholics with long alleles used alcohol more frequently during one month before admission, had much more maximum amount of beer drinking and reported withdrawal seizure more frequently than with short alleles. In contrary, alcoholics with short alleles expressed depressed mood and guilty feeling more frequently and wanted complete abstinence as a treatment goal more frequently than with long alleles. In behavioral trait, alcoholics with long alleles had higher total aggression score and showed much more self-assertive attitude(subscale of expression of aggression) than with short alleles. Allelic length of MAOA CA repeat polymorphism was correlated with self-assertive attitude and accounted for 9% of the variance of self-assertive attitude. And also, predictable variables of allelic length of MAOA CA repeat polymorphism were drinking frequency and self-assertive attitude. Our findings suggest that MAOA CA repeat polymorphism may provide some behavior modifying role especially in self-assertive attitude and indirect symptom modifying role in Korean male alcoholics.

• The Korean Journal of Mycology
• /
• v.31 no.2
• /
• pp.59-66
• /
• 2003

Laccase produced by Pycnoporus cinnabarinus SCH-3 isolated from Korea was partially purified using ultrafiltration, anion exchange chromatography and affinity chromatography, The laccase was produced as the predominant extracellular phenoloxidase during primary metabolism. Neither lignin peroxidase nor manganese-dependent peroxidase were detected in the culture fluid. In order to examine the effect of inducers in laccase production, 2,5-xylidine was added in the culture of Pycnoporus cinnabarinus SCH-3. Addition of 2,5-xylidine enhanced 25-fold laccase production. Purified laccase was a single polypeptide having a molecular mass of approximately 66 kDa, as determined by SDS-polyacrylamide gel electrophoresis, and carbohydrate content of 9%. $K_{m}\;and\;V_{max}$ values for laccase with ABTS [2,2-azinobis (3-ethylbenzthiazoline 6-sulfonic acid)] as a substrate (Lineweaver-Burk plot) was determined to be $44.4{\mu}M\;and\;56.0{\mu}mole$, respectively. The optimal pH for laccase activity was found to be 3.0. The enzyme was very stable for 1 hour at $60{\circ}C$. Half-life ($t_{1/2}$) of the enzyme was about 10 min at $80{\circ}C$. Spectroscopic analysis of purified enzyme indicated that the enzyme was typical of copper-containing protein. Substrate specificity and inhibitor studies for laccase also indicated to be a typical fungal laccase. The N-terminal amino acid sequence of the P. cinnabarinus SCH-3 laccase showed 94% of homology to the N-terminal sequences of laccases from P. cinnabarinus PB and P. coccineus.

• Restorative Dentistry and Endodontics
• /
• v.25 no.4
• /
• pp.561-574
• /
• 2000

Poly-P has been used to prevent decomposition of foods and has been shown to have inhibitory effect on the growth of gram positive bacteria. The purpose of this study was to evaluate the effect of poly-P on the growth of Porphyromonas endodontalis, a gram negative obligate anaerobic rod, endodontopathic bacterium. P. endodontalis ATCC 35406 was in BHI broth containing hemin and vitamin K with or without poly-P. Inhibitory effect of each poly-P which was added at the beginning(lag phase) or during(exponential phase) the culture, MIC(minimum inhibitory concentration) was determined by measuring the optical density of the bacterial cell at 540nm. Viable cell counts were measured to determined whether poly-P has a bactericidal effect. Leakage of intracellular nucleotides from P. endodontalis was determined at 260nm and morphological change of P. endodontalis was observed under the TEM(transmission electron microscope). Binding of 32P-labeled poly-P to P. endodontalis was examined. SDS-polyacrylamide gel electrophoresis and zymography were performed to observe the changes in protein and enzyme profiles of P. endodontalis, respectively. The results from this study were as follows : 1. The minimal inhibitory concentration(MIC) of poly-P to P. endodontalis appeared to be 0.04~0.05%. 2. Poly-P added to the P. endodontalis culture during the exponential phase of P. endodontalis was as much effective as poly-P added at the begining of the culture, suggesting that the antibacterial effect of poly-P is not much dependent on the initial inoculum size of P. endodontalis. 3. Poly-P are bactericidal to P. endodontalis, demonstrating the decrease of the viable cell counts. 4. Intracellular nucleotide release from the P. endodontalis, was not increased in the presence of poly-P and was not reversed by the addition of divalent cations like $Ca^{2+}$ and $Mg^{2-}$. 5. Under the TEM, it was observed that fine electro-dense materials were prominent in the poly-P grown P. endodontalis, appearing locally in the cell, and the materials were more abundant and more dispersed in the cell as the incubation time with poly-P increased. In addition, highly electron dense granules accumulated in many poly-P grown cells, most of which were atypical in their shape. 6. Binding of 32P-labeled poly-P to P. endodontalis appeared to be 32.8 and 45.5 and 53.4% at 30 minutes, 1 hours and 2 hours, respectively. 7. In the presence of poly-P. the synthesis of proteins with apparent molecular masses of 25, 27, 35, 45 was lost or drastically decreased whereas expression of a protein with an apparent molecular mass of 75 was elevated. 8. Proteolytic activity of P. endodontalis was decreased by poly-P. The overall results suggest that use of poly-P may affect the growth of P. endodontalis, and the anti-bacterial activity of poly-P seems largely bactericidal. Changes in shape, protein expression, and proteolytic activity of P. endodontalis by poly-P may be directly and indirectly attributed to the antibacterial effect of poly-P. Further studies will be needed to confirm the effect of poly-P.

• Journal of Korean Ophthalmic Optics Society
• /
• v.18 no.3
• /
• pp.253-260
• /
• 2013

Purpose: To investigate the UV-B blocking rate of eyeglasses lens which can prevent enzymes denaturation in cornea. Methods: The denaturation degree of RNase A and catalase, superoxide dismutase (SOD) was determined by using Acrylamide gel electrophoresis after UV-B irradiation of 312 nm for 1, 3, 6, 24 and 96 hours. Also, the inhibitory effect of eyeglasses lens having UV-B blocking rate of 50%, 80%, 95% and 99% on the enzymes denatration was measured. Results: The denaturation of RNase A was induced by 1 hour-irradiation of UV-B. To inhibit RNase A denaturation after UV-B irradiation between 1 hour and 6 hours, UV-B blocking lens of 95% were effective. UV-B blocking lens of 99% suppressed the inhibition of RNase A denaturation after the UV-B exposure between 24 hours and 96 hours. The denaturation of catalase was not induced by 1 hourirradiation of UV-B. To inhibit enzyme denaturation after UV-B irradiation between 1 hour and 6 hours, UV-B blocking lens of 50% were effective. UV-B blocking lens of 95% suppressed the inhibition of enzyme denaturation induced by UV-B irradiation between 24 hours and 96 hours. The SOD denaturation was not induced by UV-B irradiation shorter than 6 hours exposure. The UV-B blocking lens of 50% could inhibit SOD denaturation after the UV-B irradiation for 24 hours. When SOD was exposed to UV-B for 96 hrs, SOD denaturation was inhibited by eyeglasses lens with UV blocking rate higher than 95%. Conclusions: The results demonstrated that the proper UV-B blocking rates of eyeglasses lens to inhibit the enzymes denaturatioin was different according to the types of enzymes and its inhibitory effect was effective only when eyeglasses lens had higher than certain UV-B blocking rate.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.19 no.1
• /
• pp.60-66
• /
• 1986

The muscles of wild and cultured eel, Anguilla japonica, were analyzed for the protein composition and amino acid profile. The differences of the subunit distribution for the sarcoplasmic and myofibrillar proteins were discussed with sodium dodecylsulfate(SDS) polyacryamide gel electrophoresis. The muscle protein in wild eel was composed of $30.78\%$ of sarcoplasmic, $59.02\%$ of myofibrillar, $9.73\%$ of residual intracellular and $2.47\%$ of stroma fraction. That in cultured eel was composed of $31.81\%,\;58.37\%,\;8.16\%\;and\;1.80%$, respectively, The sarcoplasmic and myofibrillar proteins were composed of 16 and 14 subunits in wild eel, and 22 and 15 subunts in cured eel. The sarcoplasmic protein between wild and cultured muscles showed a similar trend in the subunits, except a few subunits such as 36,500, 46,000, 58,500, 75,000, 170,000 and 235,000 daltons in cultured eel. Only the existence of 45,000 dalton subunit was the difference between wild and cultured eel in myofibrillar protein. The distribution patterns of total amino acid in muscles of wild and cultured eel were found to be very similar trend, although glycine content in wild eel was slightly higher than that in cultured one.

• Journal of radiological science and technology
• /
• v.28 no.4
• /
• pp.333-340
• /
• 2005

In this study, the radioprotective effects of Grifola umbellata hot water extracts (Gu-extract) on mice were investigated. Single pre-administration of Gu-extract increased the 40-day survival ratio of irradiated mice from 65.5％ to 78.6％. The growth of 3 week old male mice in the irradiated group was slightly retarded as compared to those of the control and Gu-extract treated mice. The average spleen and thymus weights of the irradiated mice were lower than those of the control and Gu-extract treated mice. The weight reduction of testis in the irradiated mice was significant. While it was relatively slight in the Gu-extract treated mice as compared to that of control mice. No significant difference in the weight was observed in heart, kidney or liver among three groups. The leukocytes of the Gu-extract treated mice did not decrease dramatically as in the irradiated group, but recovery patterns were similar in both groups. Reduction of erythrocytes were similar in both groups but its recovery occurred more rapidly in the extract treated group. The glucose level of the Gu-extract treated group did not change during the period examined, while it was still higher in the irradiated group than the level in the control group in two weeks. The cholesterol levels in the irradiated and the Gu-extract treated groups were higher than that of control group on day 7, but decreased to the level of the control group on day 14. No difference was observed in total protein amount of the serum among the three groups. SDS-polyacrylamide gel electrophoresis of the soluble proteins extracted from various organs did not reveal differences to any extent in all groups except in the livers of the irradiated and extract treated groups, in which some proteins were missing or less present.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 1995.06b
• /
• pp.77-96
• /
• 1995

To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

• Food Science of Animal Resources
• /
• v.29 no.2
• /
• pp.238-244
• /
• 2009

Food allergy is a serious nutritional problem in both children and adults. Therefore, food allergenicity reduction methods are greatly needed. The allergenicity is altered by various manufacturing processes, and the digestibility of food proteins can be affected by food processing. This study was conducted to investigate the effect of in-vitro digestibility on the allergenicity of autoclaved market pork sausages using pepsin (30min) and trypsin (5, 30, 60, 90, and 120min). The binding ability of the porcine serum albumin (PSA) from sausages A and B significantly decreased by about 30 and 23%, respectively, after autoclave treatment (121; 5, 10, and 30 min). After the pepsin and trypsin treatments, the binding ability of products A and B at 30 min decreased. These competitive indirect enzyme-linked immunosorbent assay (ci-ELISA) results corresponded well with the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting results. The results demonstrated that the allergenicity of pork sausages considerably decreased after autoclave treatment, and were also maintained or decreased after enzyme treatment. Accordingly, autoclave treatment represents a promising processing technology for the reduction of the allergenicity of diverse food products.