• Title/Summary/Keyword: culture pH

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Establishment of a Dental Unit Biofilm Model Using Well-Plate (Well-Plate를 사용한 치과용 유니트 수관 바이오필름 모델 확립)

  • Yoon, Hye Young;Lee, Si Young
    • Journal of dental hygiene science
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    • v.17 no.4
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    • pp.283-289
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    • 2017
  • The water discharged from dental unit waterlines (DUWLs) is heavily contaminated with bacteria. The development of efficient disinfectants is required to maintain good quality DUWL water. The purpose of this study was to establish a DUWL biofilm model using well-plates to confirm the effectiveness of disinfectants in the laboratory. Bacteria were obtained from the water discharged from DUWLs and incubated in R2A liquid medium for 10 days. The bacterial solution cultured for 10 days was made into stock and these stocks were incubated in R2A broth and batch mode for 5 days. Batch-cultured bacterial culture solution and polyurethane tubing sections were incubated in 12-well plates for 4 days. Biofilm accumulation was confirmed through plating on R2A solid medium. In addition, the thickness of the biofilm and the shape and distribution of the constituent bacteria were confirmed using confocal laser microscopy and scanning electron microscopy. The average accumulation of the cultured biofilm over 4 days amounted to $1.15{\times}10^7CFU/cm^2$. The biofilm was widely distributed on the inner surface of the polyurethane tubing and consisted of cocci, short-length rods and medium-length rods. The biofilm thickness ranged from $2{\mu}m$ to $7{\mu}m$. The DUWL biofilm model produced in this study can be used to develop disinfectants and study DUWL biofilm-forming bacteria.

Effect of Continuous Cultivation Years on Soil Properties, Weed Occurrence, and Rice Yield in No-tillage Machine Transplanting and Direct Dry-seeding Culture of Rice (벼 무경운 기계이앙 및 건답직파 연속재배년수가 토양특성, 잡초발생 및 벼 생육에 미치는 영향)

  • Park, Hong-Kyu;Kim, Sang-Su;Choi, Won-Yong;Lee, Ki-Sang;Lee, Jae-Kil
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47 no.3
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    • pp.167-173
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    • 2002
  • This experiment was conducted to figure out the change of soil physical properties, rice growth and yield with the years of continuous cultivation in direct dry-seeding and no-tillage machine transplanting. Experiments were conducted at NHAES(National Honam Agricultural Experiment Station, RDA, Iksan, Cheon Buk Province, South Korea) with a rice variety "Dongjinbyeo" from 1995 to 2000. In no-tillage machine transplanting cultivation, organic matter in soil was higher than that on direct dry-seeding and was significantly high in topsoil. Problematic weed species were E. crus-galli B., A. keisak H., and L. japonica M. Plant height and tiller number m-2 were higher in common-tillage during the total growth duration. The highest weedy rice occurrence of 27.5% was observed in live years' continuous direct dry-seeding and followed by 6.2%, in four years', and 3.7%, in three years'. The highest yield reduction of 38% was observed in five years' continuous direct dry-seeding. The reduction may resulted from the competition between weedy rice and cultivated rice.

Development of Functional Food Materials from Acanthopanax senticosus-Fermented Mushroom Mycelia (가시오가피 버섯균사체 발효물의 기능성 식품 소재 개발)

  • Cho, Ju-Hyun;Choi, Goo-Hee;Park, In-Jae;Baik, Soon-Ok;Kim, Hyung-Ha;Kim, Choong-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.411-418
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    • 2014
  • Three mushroom mycelia, Ganoderma lucidum, Hericium erinaceum, and Phellinus linteus, were separately diluted with the natural culture media Acanthopanax senticosus. Solid-state fermentation was used to produce three different A. senticosus-fermented mushroom mycelium groups: G. lucidum mycelia, H. erinaceum mycelia, and P. linteus mycelia. The resulting mycelia were analyzed to assess their efficacies as health functional foods. Optimized fermentation conditions were determined by considering the density and growth speed of mycelia in each A. senticosus-fermented mushroom mycelium group. The cultured mushroom mycelia under the optimized conditions were extracted using water and 70% ethanol. Extraction was followed by filtration, concentration and freeze-drying to produce extract powder of A. senticosus-fermented mushroom mycelia: Water extracts (FM-5111, FM-5121, and FM-5131) and 70% ethanol extracts (FM-5112, FM-5122, and FM-5132). Analysis of extract powder of A. senticosus-fermented mushroom mycelia was performed using the maker compounds eleutheroside B and eleutheroside E. Analysis of ${\beta}$-glucan contents was performed by enzymatic procedures.

A New Synthetic Medium for Lactic Lactococci: Application to Marine lactic Acid Bacteria

  • KIM Joong K.;BAJPAI Rakesh K.
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.6
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    • pp.812-813
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    • 1995
  • Lactococcal cells are nutritionally fastidious and thus, generally cultured either in milk or M17 medium (Terzaghi and Sandine, 1975). In this study, Lactococcus cremoris wild-type (KH) and its less­proteolytic mutant (KHA1) cells were grown on the M17 medium or with modified M17 medium by replicated parallel experiments. The modified M17 medium had the same composition as M17 medium, except that lactose was replaced by glucose. Analyses of culture-broth samples, in which the M17 and the modified M17 media were used, were conducted by high-performance liquid chromatography (HPLC). But, working with these media created noisy problems in analyses of samples. Therefore, a new semi-synthetic medium was developed on the basis of nutritional requirements (Morishita et al., 1981). The composition of the semi-synthetic medium determined on the basis of the nutritional requirements and the composition of milk, is presented in Table 1. The composition of M17 medium is also presented and compared in the table. L. cremoris KH and KHA1 cells were grown again on the new synthetic medium containing glucose or lactose. The broth samples were then drawn and analyzed by HPLC. Clearer separations of fermented products were achieved from the new medium than those with the M17 and the modified M17 media. In comparison with the M17 or the modified M17 media, growth on the new medium was good (Kim et al, 1993). Additional fermentations were also carried out at a controlled pH of 7.0, where enhanced growth of lactococcal cells was obtained. In the fermentations, samples were also analyzed for the concentrations of sugar and lactic acid. The results showed that the new synthetic medium was as good as or better than the M 17 and the modified M 17 media. This is because casein hydrolysate in the synthetic medium provided a ready supply of amino acids and peptides for L. cremoris KH and KHA1 cells. Lactic acid bacteria (LAB) including Lactococcal cells have been known to be an effective means of preserving foods, at the same time as giving particular tastes in fields of dairy products. LAB also have always occupied an important place in the technology of sea products, and marine LAB have known to be present in traditional fermented products (Ohhira et al, 1988). To apply the new synthetic medium to marine LAB, two different LAB were isolated from pickled anchovy and pollacks caviar and were grown on the new media in which various concentrations of NaCl $(3, 5, 7 and 10\%)$ added. They were also grown on the medium solution in natural seawater $(35\%o\;salinity)$ and on the solution of natural seawater itself, too. As seen in Fig. 1, Marine LAB were grown best on the synthetic medium solution in natural seawater and the higher concentrations of NaCl were added to the medium, the longer lag-phase of growth profile appeared. Marine LAB in natural seawater were not grown well. From these results, the synthetic medium seems good to cultivate cells which are essential to get salted fish aged. In this study, it showed that the new synthetic medium provided adequate nutrition for L. cremoris KH and KHA1 cells, which have been used as cheese starters (Stadhouders et al, 1988). Using this new medium, the acid production capability of starter cultures could be also measured quantitatively. Thus, this new medium was inferior to the M17 or the modified M17 medium in culturing the cheese starters and in measuring fermentation characteristics of the starter cells. Moreover, this new medium found to be good for selected and well-identified marine LAB which are used in rapid fermentations of low-salted fish.

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Introduction and Expression of PAP gene using Agrobacterium in Scrophularia buergeriana Miquel (Agrobacterium을 이용한 PAP 유전자의 현삼으로 도입 및 형질발현)

  • Yu, Chang-Yeon;Seong, Eun-Soo;Lim, Jung-Dae;Huang, Shan-Ai;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.2
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    • pp.156-165
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    • 2001
  • Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

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Oocyte-sperm Binding Assay (OSBA) Technique for Rapid Q/C of IVF Culture Condition (체외수정용 배양조건의 신속한 Q/C를 위한 정자-난자 결합분석법(OSBA) 개발)

  • 정구민;신영수
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.163-169
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    • 2001
  • OSBA(oocytes-sperm binding assay) is a tool developed for rapid test of optimal condition of IVF medium and protein source by binding ability of mouse sperm and egg. Mouse oocyte-cumulus complexes were prepared by removing of the cumulus cells with 0.1% hyaluronidase. 10$\pm$2 oocytes per 30 ${mu}ell$ medium drop were inseminated with 3 ${mu}ell$ sperm suspension and were cultured f3r 3 hours and 24 hours, respectively. And the oocytes were recovered gently and the No. of sperm bound on oocytes were counted. In the Exp. 1, the ratio of oocytes bound with one sperm at least were 60.2%(50/83), 2%(2/77) and 100%(79/79) in the medium with no protein, FBS(15%, v/v) and BSA(0.4%. w/v), respectively, Fetal bovine serum(FBS) seriously inhibited sperm binding on oocyte, although bovine serum albumin(BSA) promoted the binding ability. The inhibiting effect of FBS was dependent on the concentration of FBS. The sperm binding ability according to oocyte maturity was tested in the Exp. 2. There was no significant difference between Met. II (mature) and Met. I (intermediate mature) oocytes in the number of oocytes bound with sperm and the number of sperm bound on oocytes. Finally, in Exp. 3, two batches of Ham's F10 medium with good and poor quality by OSBA were tested (The ratios of embryos developed from PN 1-cell stage to hatched blastocyst; 25% vs. 70%). In the medium with good quality, sperm binding ability was significantly increased (P < 0.05). The ratio of oocytes bound with one sperm at least was 66% and 90% in the medium with poor and good quality, respectively. Conclusively, It was possible to test IVF medium condition rapidly and easily by OSBA.

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Uptake of Heavy Metal Ions by Water Dropwort (Oenanthe stolonifera DC.) and Identification of Its Heavy Metal-Binding Protein (미나리의 중금속 흡수량 측정 및 중금속 결합단백질의 동정)

  • Park, Young-Il;Kim, Hee-Guen;Kim, Yoo-Young;Kim, In-Soo
    • Applied Biological Chemistry
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    • v.39 no.6
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    • pp.494-500
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    • 1996
  • Uptake of hen metal ions by water dropwort (Oenanthe stolonifera DC.) and its cadmium-binding protein were studied to probe for good method to remove heavy metal contaminants from environments. The plant was cultured in the culture medium (pH 7.0) containing the various concentrations of $Cd^{2+}$, $Cr^{3+}$ or $Pb^{2+}$, for 3 and 7 days. The residual heavy metals deposited in roots linearly increased as the metal ions concentration increased up to 17 ppm for $Cd^{2+}$, 20 ppm for $Cr^{3+}$ and 50 ppm for $Pb^{2+}$. Above these concentrations, the plant growth was inhibited and the uptake rates of the metal ions decreased. The heavy metals absorbed by the plant were mostly deposited in roots. In particular, the residual concentration of lead in roots was about four times higher than those of cadmium and chromium. When cultured in the medium containing 20 ppm of each metal ion, 80% of cadmium, 90% of cromium and 96% of lead were deposited in roots out of the total residual metal ions in the plant. These values correspond to 6.1 mg of cadmium, 5.2 mg of chromium and 23.6 mg of lead per one gram of roots tissue on a dry weight basis. A cadmium-binding protein was partially purified by extraction, gel filtration and DEAE-Cellulose chromatography from water dropworts that was grown in the medium containing 20 ppm $Cd^{2+}$. The purified protein was a single band on SDS- and non-denaturing- polyacrylamide gel electrophoresis. Its molecular mass was estimated to be ca. 5,000 dalton by gel filteration. Analysis of amino acid composition of the protein indicated that it had a typical amino acid composition of heavy metal-binding protein in that it contained 27% of acidic amino acids and 9.9% of cysteine. However, it is likely that the protein is a new plant metal-binding protein, since its amino acid composition is somewhat different from those of phytochelatins that have been known so far.

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Expression and Purification of Recombinant Human Interferon-gamma Produced by Escherichia coli (대장균이 생산한 재조합 인체 감마인터페론의 발현과 정제)

  • Park, Jung-Ryeol;Kim, Sung-Woo;Kim, Jae-Bum;Jung, Woo-Hyuk;Han, Myung-Wan;Jo, Young-Bae;Jung, Joon-Ki
    • KSBB Journal
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    • v.21 no.3
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    • pp.204-211
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    • 2006
  • For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

The Role of Factors Controlling the Accumulation of Biogenic Amines in Various Cheeses as Milk-Based Products: A Review (낙농유제품인 치즈에 축적된 생체 아민의 다양한 영향 인자에 관한 연구: 총설)

  • Chon, Jung-Whan;Kim, Dong-Hyeon;Kim, Hyun-Sook;Song, Kwang-Young;Lim, Jong-Soo;Choi, Dasom;Kim, Young-Ji;Lee, Soo-Kyung;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.32 no.2
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    • pp.77-92
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    • 2014
  • Fermented foods have often been implicated as causative agents in poisoning due to toxic levels of biogenic amines. Cheese, a milk-based fermented food, is the product most likely to contain potentially harmful levels of biogenic amines, such as tyramine, histamine, putrescine, and so on. Recently, the risk awareness of a dietary uptake of high loads of biogenic amines has increased. Hence, we here review the published literature on several factors known to affect the biosynthesis of biogenic amines and their accumulation in milk-based foods. Furthermore, with regard to risk analysis, we discuss the control of factors related to the synthesis and accumulation of biogenic amines as a means to reduce their incidence in milk-based products, and thus to increase food safety.

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Economic Analysis, Growth and Pests of Wheat (Triticum aestivum L.) in Gelatin·Chitin Microorganisms-treated Organic Culture (젤라틴·키틴분해미생물을 이용한 밀 유기재배와 관행재배의 생육, 병해충 발생조사 및 경제성 분석)

  • Ahn, Philip;Lee, Jiho;Cha, Kwang-Hong;Seo, Dong-Jun;An, Kyu-Nam;Yoon, Chang-Yong;Kim, Kil-Yong;Jung, Woo-Jin
    • Korean Journal of Organic Agriculture
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    • v.29 no.2
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    • pp.223-240
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    • 2021
  • This study was carried out to investigate the economic value of organic wheat production using gelatin·chitin microorganisms in Gwangsan-gu, Gwangju city. The soil condition of experiment field was clay loam Jisan series. The organically cultivated fields were sprayed gelatin and chitin degrading bacteria. The test was performed at conventionally cultivated field and organically cultivated field. Emergence of weed on organically cultivated field was significantly higher than conventionally cultivated field which sprayed herbicide before seeding. Weed emergence have a critical impact on grain yield. Occurrence of diseases and insect pests were higher than conventionally cultivated fields. In 2019, the amount of lodging in conventionally cultivated field were higher than conventionally cultivated field. In 2020, lodging and wet injury were occur in both field. Comparing yield element between organically and conventionally cultivated experimental area, grain yield in organically cultivated field was shown slightly higher amount than conventionally cultivated field. However in the actual yield of 2019, organically cultivated field shows 20% deceased yield because of overgrown weed. In 2020, weed emergence and yellow mosaic virus by wet injury cause 30% decease in the grain yield in organically cultivated field. Content of protein, carbohydrates, ash, water and fat in the grain were not different significance. In 2019, net incomes of conventionally cultivated wheat was 461,031 won/0.1 ha while organically cultivated wheat was 443,437 won/0.1 ha. In the rate of income, conventionally cultivated field was 83.0% as against organically cultivated field (73.3%). In 2020, net incomes of organically cultivated wheat was 437,812 won/0.1 ha while conventionally cultivated wheat was 418,281 won/0.1 ha. In the rate of income, conventionally cultivated field was 81.6% as against organically cultivated field (73.0%).