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Strain Improvement of Leuconostoc mesenteroides as a Acid-Resistant Mutant and Effect on Kimchi fermentation as a Starter (Leuconostoc mesenteroides의 내산성 변이주의 김치발효에 미치는 효과)

  • Kim Young-Hwan;Kim Hee-Zoong;Kim Ji-Young;Choi Tae-Bu;Kang Sang-Mo
    • Microbiology and Biotechnology Letters
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    • v.33 no.1
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    • pp.41-50
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    • 2005
  • An organic acid tolerance mutant (M-200) was obtained from Leuconostoc mesenteroides KCCM 35471, followed by the screening procedure using a specific organic acid medium (lactic acid: acetic acid, 2:1). The characteristics of the acid tolerance M-200 and the wild type LM-W were examined at various temperature and pH ranges $(l0-30^{\circ}C$ of temp, 3.5-4.5 of pH). The growth of strain M-200 at HCl adjusted medium $(10^{\circ}C\;and\;pH 3.5)$ was observed. In the case of organic acid adjusted medium, the strain showed its growth at the pH range of 3.8. When the strain M-200 was used as a starter for Kimchi fermentation, a constant acid level (0.55) was observed during the whole fermentation period. This result indicates that the strain produces a proper level of acid content for the Kimchi fermentation. This result also indicates that the edible period of Kimchi can be extended to 3.5 fold compare to the result obtained from the LM-W used Kimchi fermentation. However the excess use of the strain M-200 showed the inhibition of growth of Lactobacillus plantarum, low lactic acid level content and low level of organoleptic test. In the case of organic acid content during the Kimchi fermentation, the strain M-200 showed relatively low production rate compare to the wild type (M-200: 3.5 mg/L at 21 days of fermentation, LM-W: 7 mg/L at 21 days of fermentation). Therefore a mixed Kimchi starter containing M-200 and other strains probably maintain a good Kimchi quality during the fermentation.

Development of pSJE6c, an Expression Vector for Kimchi Lactic Acid Bacteria, and Heterologous Gene Expression Using the Vector (김치유산균용 발현벡터 pSJE6c 개발과 이를 이용한 외래 유전자 발현)

  • Lee, Kang-Wook;Park, Ji-Yeong;Lee, Ji-Yeon;Lee, Hwang-A;Baek, Chang-Un;Jo, Hyeon-Deok;Kim, Joo-Yeon;Kwon, Gun-Hee;Chun, Ji_Yeon;Kim, Jeong-Hwan
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.389-398
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    • 2009
  • Development of expression vectors is important for the basic and applied researches on kimchi LAB (lactic acid bacteria). An expression vector, pSJE6c was constructed by inserting P6C promoter sequence from Lactococcus lactis into pSJE, a shuttle vector for E. coli and Leuconostoc species. To test the efficiency of pSJE6c, aga ($\alpha$-galactosidase) and lacZ ($\beta$-galactosidase) genes were expressed in Lactobacillus brevis 2.14. Compared to the pSJE, expression levels of both genes were increased, indicating P6C promoter was better than indigenous promoters. Enzyme activities of L. brevis cells harboring pSJE6caga (pSJE6c with aga) or pSJE6Z (pSJE6c with lacZ) were 1.5-2 fold higher than those with pSJEaga (pSJE with aga) or pSJEZ (pSJE with lacZ). More RNA transcripts were detected in cells harboring pSJE6c based recombinant plasmid. The results indicated that heterologous gene expressions in kimchi LAB could be improved significantly by use of efficient expression vectors.

A Study on Causal Relationship About the Reparations Range (손해배상범위에 관한 인과관계의 연구)

  • Choi Hwan-Seok;Park Jong-Ryeol
    • The Journal of the Korea Contents Association
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    • v.6 no.4
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    • pp.146-157
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    • 2006
  • Causal relationship means what relations the result occurred have with a fact as a reason. In general, a formular that no result exists without reasons is used for the method to confirm existence and inexistence of causal relationship. Problematic causal relationships in Private Law are reparations (Article No. 393 of Private Law) due to debt nonfulfillment and reparation due to tort (Application of Article No. 393 by Article No. 750, and No. 763 of Private Law). The purpose pursued by reparation system in private law is to promote equal burden of damages, and the range of reparation at this time is decided by the range of damage and the range of damage is decided by the principle of causal relationship. That the causal relationship theory fairly causes confusion by treating one problem and the other problem as the same thing, instead of dividing them according to the purpose of protection presented by the law is a reason of the criticism from different views.

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The Distribution of ${\gamma}{\delta}$ T Cells in Tuberculous Lymphadenopathy (결핵성 림프절에서 ${\gamma}{\delta}$ T 림프구의 분포에 관한 연구)

  • Shim, Tae-Sun;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.5
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    • pp.484-488
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    • 1994
  • Background : The antigen-specific receptor on the surface of most peripheral T lymphocytes is a disulfide-linked heterodimer composed of $\alpha$ and $\gamma$ subunits, noncovalently associated with CD3 polypeptides. Recently, a novel type of CD3-associated heterodimer was described on a T cell subset that does not express CD4 or CD8 molecules. This second type of TCR dimer is composed of chains encoded for by the $\gamma$- and $\delta$-TCR genes. These cells may exert both cytotoxic and lymphokine producing functions. Although it was reported that some ${\gamma}{\delta}$-TCR might recognize an MHC-linked determinant, the funεtion or physiologic ligand for this new receptor is not yet clear. It was found that ${\gamma}{\delta}$-TCR can react with 65 kD heat shock protein of M. tuberculosis, which suggests the possible protective role of ${\gamma}{\delta}$ T lymphocytes against tuberculosis. In our previous study, there was neither the increase in number nor the functional activation of ${\gamma}{\delta}$ T cells in the peripheral blood from patients with pulmonary tuberculosis. Now we report the distribution of ${\gamma}{\delta}$ T cells in the regional sites of M. tuberculosis infection, especial1y tuberculous lymphadenitis. Methods : Lymph nodes from patients with pathologically-proven tuberculous lymphadenopathy (n=5) and reactive hyperplasia (n=3) were used. Tissues were frozen in liquid nitrogen immediately after removal and stored below $-70^{\circ}C$. The cryostat sections of these frozen specimens were stained with anti-Leu-4 Ab, Identi-T TCR ${\delta}1$, and Identi-T ${\beta}F1$. The number of positively stained cells were counted at high power field. Results : The infiltration of ${\gamma}{\delta}$ T cells was significantly higher in the lymph nodes from patients with tuberculous lymphadenopathy than that with reactive hyperplasia ($16.3{\pm}10.3%$ vs. $1.7{\pm}1.5%$). Conclusion : These results suggest that ${\gamma}{\delta}$) T cells may play a role in the defense against M. tuberculosis infection, especially in the regional sites of infection.

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A Case of Kabuki Syndrome Confirmed by Genetic Analysis: A Novel Frameshift Mutation in the KMT2D Gene (분자유전학적으로 진단된 가부키 증후군 1례)

  • Park, Su Jin;Ahn, Moon Bae;Jang, Woori;Cho, Won Kyung;Chae, Hyo Jin;Kim, Myung Shin;Suh, Byung Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.17 no.3
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    • pp.103-108
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    • 2017
  • Kabuki syndrome is a rare congenital disorder that causes multiple birth defects and mental retardation. Mutation of the lysine methyltransferase 2D (KMT2D) gene is the primary cause of Kabuki syndrome. We report a 4-year-old Korean girl diagnosed with Kabuki syndrome based on distinctive facial features (eversion of the lower lateral eyelid, arched eyebrows, depressed nasal tip, prominent ears), skeletal anomalies, short stature, and molecular analysis, which revealed a novel frameshift mutation in the KMT2D gene. A 4-year-old patient had a past history of congenital cardiac malformations (coarctation of the aorta, ventricular septal defect, atrial septal defect, patent ductus arteriosus), subclinical hypothyroidism and dysmorphic features at birth including webbed neck, short fingers, high arched palate, micrognathia and horseshoe kidney. She showed unique facial features such as a long palpebral fissure, long eyelashes, arched eyebrows with sparseness of the lateral third, broad nasal root, anteverted ears, and small mouth. Her facial features suggested Kabuki syndrome, and genetic analysis discovered a novel heterozygous frameshift mutation (c.4379dup, p.Leu1461Thrfs*30) in exon 15 of the KMT2D gene. The diagnosis of our 4-year-old patient was made through thorough physical examination and history taking, and genetic testing. It is challenging to diagnose patients with Kabuki syndrome at birth, since the characteristic facial features are expressed gradually during growth. Clinical suspicion aroused by regular follow-ups may lead to earlier diagnosis and interventions.

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Development of animal protein(feed for fry) utilizing the rumen ciliates (제1위 섬모충(rumen ciliates)을 이용한 동물성 단백질(치어용 사료) 개발)

  • Jee, Cha-ho;Hyun, Gong-yool
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.327-336
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    • 1995
  • This study was carried out to develop the animal protein(feed for fry) that was isolated, purified and lyophilized the rumen ciliates from the healthy rumen contents which have $10^5-10^6/g$ ciliates and were discarded in abattoirs. The rumen ciliates are non-pathogenic, anaerobic and the weight of this protozoa is 2% of rumen content. The rumen protozoan and bacterial proteins both have a biological value for rats of 80-81, which is higher than the 72 of brewer's yeasts. Furthermore, the true digestibility and net protein utility of the protozoan protein are 91 and 73, much higher than those of bacterial(74 and 60) or yeast(84 and 60) proteins. The amino acids of rumen protozoa is nutritionally superior than the others. The size of rumen ciliates is $30-200{\times}20-110{\mu}m$ and so we had isolated and purified the rumen ciliates from the rumen contents by the physical methods. The purified rumen protozoa was lyophilized with freezing dryer. The results of this experiment were as follows : 1. Population dynamics of protozoan ciliates in slaughtered rumens; % of samples which small ciliates were predominated was 82.5%(52/63) and that of large ciliates was 17.5%(11/63). 1) predominant species of small ciliates were Entodinium ovinum and E nanellum. 2) predominant species of large ciliates were Epidinium ecaudatum and Diploplastron affine. 2. The lyophilized rumen ciliates which were isolated and purified from 1 kg of rumen content at the pH 6.2-6.8 was about 7.0 gram. 3. The nutrient analysis of lyophilized rqmen ciliates(LRC) was as follows: 1) Proximate analysis of the LRC and the composition of fry feed; moisture 8.05%(below 10.0), protein 35.37%(45), fat 5.39%(4.5), fiber 1.23%(below 2.5), ash 2.25%(below 15.0), Ca 0.26%(below 2.0), P 0.14%(below 1.1), energy 4,608.11(fish meal 5000 cal/g) 2) Amino acids (% in crude protein) of the LRC and the rotifer(Brachionus plicatilis); Arg 5.19%(4.50), His 2.50%(1.55), Ile 5.29%(3.45), Leu 8.11%(5.85), Lys 10.34%(6.15), Met 2.25% (0.85), Phe 5.66%(3.80), Thr 5.14% (3.45), Val 4.18%(3.90), Ala 4.13%(3.35), Asp 13.26%(8.25), Glu 16.62%(9.20), Gly 4.23%(3.10), Pro 3.25%(5.05), Ser 4.85%(3.85), Tyr 5.04%(3.05) 3) Fatty acids(% in fat) of the LRC and the rotifer(biological feed ; Brachionus plicatilis); myristic acid(C14:0) 3.27%(0.3), myristoleic acid(C14:1) 0.83%(-), palmitic acid(C16:0) 39.11% (23.5), palmitoleic acid(C16:1) 2.81%(2.0), stearic acid(C18:0) 9.36%(5.6), oleic acid(C18:1) 25.54%(3.5), linoleic acid(C18:2) 15.05%(32.9), linolenic acid(C18:3) 1.74%(9.8). Judging from the above investigated results, the analytical data of proximate analysis, amino acids, fatty acids of the purified and lyophilized rumen protozoa are reasonable for the feed of freshwater fishes(fry and fingerling). But it was disappointed of our expectation that the crude protein of lyophilized rumen ciliates contains low percentage, it was thought that because of the small ciliates(starch digester) in beef cattle rumens which were administered the concentrated feed, is much difficult to isolate and purify than the large ciliates(fiber digester).

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Processings of Flavoring Substances from tow-Utilized Shellfishes (연안산 저활용 패류를 이용한 풍미소재의 개발)

  • OH Kwang-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.6
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    • pp.791-798
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    • 1998
  • To develop natural flavoring substances. optimal hydrolysis conditions for two stage enzyme hydrolysates (TSEH) using low-utilized shellfishes such as purplish clam and frozen oyster stored at $-20^{\circ}C$ for 60 days. The optimal conditions for TSEH method were revealed in temperature at $50^{\circ}C$ 3 hours digestion with alcalase (Aroase AP-10, $0.3%$ w/v, pH 8.0) at the 1st stage and $45^{\circ}C$ 2 hours digestion with neutrase (Pandidase NP-2, $0.3\%$ w/v, pH 6.0) at the 2nd stage. Among water extracts, autolytic extracts and 4 kinds of enzyme hydrolysates tests, TSEH method was superior to other methods on the aspect of yields, nitrogen contents, taste such as umami and control of off-flayer formation, and transparency of extracts. From the results of chemical experiments and sensory evaluation, we may conclude that TSEH from low-utilized marine products is more flavorable compared the conventional enzyme hydrolysates, it could be commercialized as the seasoning substances.

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A Study on Wine-Making with Dried Persimmon Produced in Korea (곶감주 개발에 관한 연구)

  • Woo, Kang-Lyung;Lee, Su-Hak
    • Korean Journal of Food Science and Technology
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    • v.26 no.3
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    • pp.204-212
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    • 1994
  • To estimate the possibility of wine-making with Korean dried persimmon, its homogenized and filtered solution was fermented at $15^{\circ}C$ and $25^{\circ}C$ for 12 weeks with Saccharomyces cerevisiae (Japan Alcoholic Beverage Association N0.7). Sugars of dried persimmon were mainly composed of 27.02% of glucose, 19.81% of fructose and 5.12% of mannose. In the fermentation at $25^{\circ}C$, glucose was almost completely consumed in 8 days, but fructose and mannose were consumed up to 64% and 74%, respectively, in the same period and were not utilized any more afterwards. In the fermentation at $15^{\circ}C$, 75% of glucose, 20% of fructose and 49% of mannose were consumed in 8 days and these sugars were continuously utilized for 12 weeks. Organic acids in the homogenized and filtered solution were levulinic acid (148.6 mg%), 4-methylvaleric acid (73.5 mg%), oxalic acid (28.7 mg%), acetic acid (8.5 mg%), N-butyric acid (8.4 mg%) and succinic acid (6.7 mg%). Irrespective of fermentation temperature, levulinic acid rapidly reduced according to progression of fermentation. Oxalic acid, N-butyric acid and succinic acid decreased at 2nd day of fermentation, and then increased at 4th and 6th days and subsequently decreased again under the levels of the solution. Acetic acid and 4-methylvaleric acid increased with the proceeding of fermentation and at 12th week of fermentation these contents were more than those of the solution. The contents of total free amino acid significantly reduced at 2th day of fermentation and then increased to the level of the solution at 12th week irrespective of fermentation temperature. Ethanol content rapidly increased to the levels of 5.3(v/v) at $15^{\circ}C$ and 9.4%(v/v) at $25^{\circ}C$ to 8th day after fermentation, but at 12th week its content was 14.5%(v/v) at $15^{\circ}C$ and 9.4%(v/v) at $25^{\circ}C$. The higher alcohots identified were 2-methyl-l-propanol, 3-methyl-ibutanol, 2-methyl-l-butanol and 2-methyl-2-propanol and the range of those contents was from 0.001% (v/v) to 0.06%(v/v). The color of the wine fermented at $15^{\circ}C$ was slightly superior but flavor and taste were slightly superior in the wine fermented at $25^{\circ}C$.

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Microwave Vacuum Drying of Germinated Colored Rice as an Enzymic Health Food (효소식품으로서 발아유색미의 마이크로파 진공건조)

  • Kim, Suk-Shin;Kim, Sang-Yong;Noh, Bong-Soo;Chang, Kyu-Seob
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.619-624
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    • 1999
  • This work was to study the potential health food use of germinated colored rice after germinating and drying using microwave under vacuum. Colored rice was soaked in water at $15^{\circ}C$ for 2 days and then germinated at $25^{\circ}C$ for $3{\sim}4\;days$. The germinated colored rice was dried by different drying methods: microwave vacuum drying 1, microwave vacuum drying $2\;(drying{\rightarrow}crushing{\rightarrow}drying)$, hot air drying, vacuum drying and freeze drying. Each drier except freeze drier was set to maintain the sample temperature at $60^{\circ}C$. During microwave vacuum drying 1 or 2, the sample reached $60^{\circ}C$ much faster (within 5 min) and was dried much faster ($2{\sim}3\;hrs$ than the other drying methods. The initial drying rate of microwave vacuum drying was ten times faster than that of hot air drying. The microwave vacuum drying 2 retained the highest ${\alpha}-amylase$ activity, followed by microwave vacuum drying 1, freeze drying, vacuum drying, and hot air drying.

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Fermented Extracts of Korean Mistletoe with Lactobacillus (FKM-110) Stimulate Macrophage and Inhibit Tumor Metastasis (유산균으로 발효된 한국산 겨우살이 추출물의 Macrophage 자극에 의한 면역학적 활성화와 종양전이 억제효과)

  • Yoon, Taek-Joon;Yoo, Yung-Choon;Kang, Tae-Bong;Lee, Kwan-Hee;Kwak, Jin-Hwan;Baek, Young-Jin;Huh, Chul-Sung;Kim, Jong-Bae
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.838-847
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    • 1999
  • Based on the results that the extract of Korean mistletoe (KM-110) has immunological and anti-tumor activities and its main component is lectin called KML-U, this study was carried out to investigate the immunostimulatory and anti-tumor activities of FKM-110, fermented KM-110 with lactobacillus, as a basic study for the development of functional food with anti-tumor activity. The amount of lectin after fermentation determined by ELISA was varied with the fermentation time and kinds of lactobacillus. Cytotoxic effects of FKM-110 on the various tumor cells was significant and dependent on the concentration of KML-U and the kinds of lactobacillus. FKM-110 stimulated macrophage and resulted in the secretion of some cytokines such as IL-1 and $IFN-{\gamma}$, but this effect was not correlated with the concentration of lectin. FKM-110 fermented with Marshall Lactobacillus casei showed the most potent antitumor activity in experimental and spontaneous metastasis models. When yoghurt produced with KM-110, Marshall Lactobacillus casei and skim milk was administered orally to mouse, the metastasis of tumor cells was significantly inhibited.

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