• Title/Summary/Keyword: storage of bacterial cells

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Effect of Silk Fibroin Biomaterial Coating on Cell Viability and Intestinal Adhesion of Probiotic Bacteria

  • Kwon, Gicheol;Heo, Bohye;Kwon, Mi Jin;Kim, Insu;Chu, Jaeryang;Kim, Byung-Yong;Kim, Byoung-Kook;Park, Sung Sun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.4
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    • pp.592-600
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    • 2021
  • Probiotics can be processed into a powder, tablet, or capsule form for easy intake. They are exposed to frequent stresses not only during complex processing steps, but also in the human body after intake. For this reason, various coating agents that promote probiotic bacterial stability in the intestinal environment have been developed. Silk fibroin (SF) is a material used in a variety of fields from drug delivery systems to enzyme immobilization and has potential as a coating agent for probiotics. In this study, we investigated this potential by coating probiotic strains with 0.1% or 1% water-soluble calcium (WSC), 1% SF, and 10% trehalose. Under simulated gastrointestinal conditions, cell viability, cell surface hydrophobicity, and cell adhesion to intestinal epithelial cells were then measured. The survival ratio after freeze-drying was highest upon addition of 0.1% WSC. The probiotic bacteria coated with SF showed improved survival by more than 10.0% under simulated gastric conditions and 4.8% under simulated intestinal conditions. Moreover, the cell adhesion to intestinal epithelial cells was elevated by 1.0-36.0%. Our results indicate that SF has positive effects on enhancing the survival and adhesion capacity of bacterial strains under environmental stresses, thus demonstrating its potential as a suitable coating agent to stabilize probiotics throughout processing, packaging, storage and consumption.

Expression and purification of Soybean $\beta$-Conglycinin from ($\beta$-Conglycinin의 대장균 발현과 정제)

  • 노영희
    • The Korean Journal of Food And Nutrition
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    • v.12 no.2
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    • pp.184-190
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    • 1999
  • Soybean protein consists of two major components $\beta$-conglycinin and glycinin which together consti-tute 70% of the total seed storage protein at maturity. $\beta$-Conglycinin is trimeric glycoprotein and for-med by the assembly of various combinations of three subunits $\alpha$,$\alpha$' and $\beta$ which have molecular weig-hts of 69,000, 72,000 and 42,000, respectively. Recently $\beta$-conglycinin was identified as powerful LDL lip-oprotein receptor activation hypercholesterolemia and major allergenic proteins. To investigate these reasons we constructed an expression system of cDNA encoding $\alpha$-subunit of $\beta$-conglycinin in Escherichia coli and purified the expressed protein. The pro-$\beta$-conglycinin synthesized in Escherichia coli BL 21 (DE3)comprised approximately 15% of the total bacterial proteins and the expressed protein are formed sol-uble and trimer such as native protein in Escherichia coli cells. The highly expressed protein was purified to homogeneity by salt precipitation with 20~40 % ammonium sulfate ion-exchange chromatography with Q-sepharose and hydrophobic column chromatography with Butyltoyopearl.

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Hygienic Quality and Genotoxicological Safety of Gamma Irradiated Pork (감마선조사에 의한 돈육의 위생화 및 유전독성학적 안전성 평가)

  • 강일준;윤정한;강영희;이효구;변명우
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.5
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    • pp.1092-1098
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    • 1999
  • Gamma irradiation was applied to pork for improving its hygienic quality and evaluating its possible genotoxicity. The effective dose of irradiation was 3 kGy in pork for the sterilization of all contaminated microorganisms tested. After 8 weeks of storage at 5oC, no growth of microorganisms except for psychrophile and total aerobic bacteria was observed in the more than 3 kGy irradiated pork. The genotoxicity of high dose irradiated pork(30 kGy) was evaluated by Salmonella typhimurium reversion assay, chromosomal aberration test and in vivo micronucleus assay. The results were negative in the bacterial reversion assay with S. typhimurium TA98, TA100, TA1535, TA1537. In chromosomal aberration tests with CHL cells and in vivo mouse micronucleus assay, no significant difference in the incidences of chromosomal aberration and micronuclei was seen between nonirradiated and 30 kGy irradiated porks. These results indicate that 30 kGy irradiated pork did not show any genotoxic effects under these experimental conditions.

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Hygienic Quality and Safety of Gamma Irradiated Angelicae Gigantis Radix and Aloe (감마선조사에 의한 당귀와 알로에의 위생화 및 안전성 평가)

  • 강일준;이수용;이상준;김광훈;이병훈
    • Toxicological Research
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    • v.13 no.1_2
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    • pp.55-60
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    • 1997
  • Gamma irradiation was applied to Angelicae gigantis radix and Aloe to improve their hygienic quality. The effective dose of irradiation was 7 kGy in Angelicae gigantis radlx and 5 kGy in Aloe for the sterilization of all contaminated microorganisms tested. After 8 months of storage at room temperature, no growth of microorganisms was observed in the irradiated products. The safety of these products were evaluated by Salmonella typhimurium reversion assay and in vivo micronucleus assay using mouse bone marrow cells. They were negative in the bacterial reversion assay with S. typhimurium TA 98, TA100, TA1535 and TA1537. In the in vivo mouse micronucleus assay, they did not show any clastogenic effect at all doses tested. These results indicate that the gamma irradiation of Angelicae gigantis radix at 12 kGy and of Aloe at 10 kGy have no genotoxic effects under these experimental conditions.

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Effect of Combined Use of Anti-microbial Materials on Storage of Low Salted Kochujang (저식염 고추장 저장시 항균물질 혼합첨가의 영향)

  • Han, Sun-Mi;Kim, Dong-Han
    • Applied Biological Chemistry
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    • v.51 no.4
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    • pp.281-287
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    • 2008
  • Effect of combined use of anti-microbial materials, such as alcohol, mustard and chitosan, or pasteurization on the quality of low salted kochujang was investigated during storage at $30^{\circ}C$ for 12 weeks. Activity of amylase decreased during storage, with lower activity in pasteurized kochujang than the other groups. Acidic protease activity increased during storage, but neutral protease activity decreased after 4 weeks. Viable cells of yeast increased during storage, but bacterial counts decreased gradually and did not show any remarkable difference among the test groups. Hunter a-values decreased as storage time increased, whereas L- and b-values decreased after 4 weeks and the degree of increase in total color difference (${\Delta}E$) was low in the supplementary ingredients added kochujang. The moisture contents and water activities decreased during storage with being lower in supplementary ingredients added groups. Titratable acidity of kochujang was decreased after 4 weeks of storage with the highest in combination of the supplementary ingredients added group. Oxidation-reduction potential was low in the supplementary ingredients added kochujang. Total sugar and reducing sugar contents of kochujang decreased during storage, with the highest contents in the supplementary ingredients added group. Ethanol content of kochujang increased during storage, whereas ethanol production was reduced in ethanol added one. Amino-nitrogen and ammonia-nitrogen contents decreased during storage with being lower in kochujang prepared with supplementary ingredients. Therefore, supplementary ingredients added kochujang would be effective for extending shelf-life of kochujang.

Development of biological agent seeded on fine sand for control of brown patch and Pythium blight disease on golf course grasses.

  • Hur, Jae-Seoun;Lim, Kwang-Mi;Oh, Soon-Ok;Yum, Kyu-Jin;Koh, Young-Jin
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.88.1-88
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    • 2003
  • Antogonistic bacteria against Rhizoctonia solani and Pythium spp., causing serious damage to golf course grasses, were isolated from the top soil of several golf courses in Korea. The isolate of Limk0102 was selected as the biological agent by characterization of antifungal activity, large scale preparation, fungicides tolerance and ecological fitness to the targe environment. The isolate was identified as Bacillus subtilis by biochemical and physiological characterization, and 165 rDNA sequence analysis. The bacterial agent was formulated as a granule type by seeding it on fine sand. The formulated agent showed high recovery rate (more than 10$\^$8/ cells/g sand) even after 6 month-storage at room temperature with similar antifungal activity with that of original cells. In vitro, the biological agent successfully exhibited antagonistic performance on bentgrass inoculated with R. solani or Pythium spp. isolated from the diseased grasses on golf courses. Field evaluation on disease control activity and ecological fitness of the agent is now under going on several golf courses.

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A Study on Stability of Nitrile Hydratase of Brevibacterium sp. CHI Under the Various Conditions (여러가지 조건하에서 Brevibacterium sp. CH1의 Nitrile Hydratase의 안정성)

  • 황준식;장호남
    • Microbiology and Biotechnology Letters
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    • v.18 no.1
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    • pp.56-60
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    • 1990
  • A bacterial strain of Brevibacterium sp. CHI was isolated from soil and used to produce an enzyme (nitrile hydratase) necessary for carrying out the bioconversion of acrylonitrile to acrylamide. Various immobilization methods and enzyme stability were investigated. The nitrile hydratase showed the maximum stability at pH 7 for the free cells. EDTA and phenyl methyl sulfonyl fluoride were selected as the protease inhibitor and the enzyme stability was evaluated by changing inhibitor concentration. Acrylamide beads were the best carriers among four carriers we tested in terms of stability and physicoehemical strength. The storage stability of the immobilized cells decreased with increasing acrylamide concentration of the gel phase at 4$^{\circ}C$, and was very low at acrylarnide concentration above 25%.

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Viability and Luciferase Activity of Freeze-Dried Recombinant Biosensor Cells for Detecting Aromatic Hydrocarbons

  • Kim, Mi-Na;Park, Hoo-Hwi;Lim, Woon-Ki;Shin, Hae-Ja
    • Biomedical Science Letters
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    • v.9 no.4
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    • pp.195-201
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    • 2003
  • Aromatic hydrocarbons are of major concern among genotoxic chemicals due to their toxicity and persistence. Some microorganisms can utilize aromatic hydrocarbons as carbon and energy sources by inducing expression of catabolic operon(s). The XylR regulatory protein activates transcription of the catabolic enzymes to degrade BTEX (benzene, toluene, ethylbenzene, and xylene) from its cognate promoters, Pu and Ps upon exposure of the cells to the aromatic hydrocarbons. The activity of XylR on the promoters was previously monitored using luciferase luc reporter system. The xylR, its promoter Pr and the promoter Po for the phenolic compound catabolic operon were introduced upstream of firefly luciferase luc in the pGL3b vector to generate about 7.1 kb of pXRBTEX. Here E. coli harboring the plasmid was freeze-dried under various conditions to fin,d optimal conditions for storage and transport. The cell viability and luciferase activity were maintained better, when the cells were freeze-dried at -7$0^{\circ}C$ in the addition of the 10% skim milk or 12% sucrose. However, coaddition of protectants such as 10% skim milk plus 10% glucose or 12% sucrose plus 10% glucose, resulted in much better viability and bioluminescence activity compared with the effect of single addition of each protectant. In addition, it was shown that the freeze-dried cells maintained almost intact bioluminescent activities and cell viability for at least 1 week after freeze-drying. This work demonstrated that the properly freeze-dried recombinant bacterial cells could be utilized as a whole-cell biosensor for simple and rapid monitoring of BTEX in the environment.

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Shelf-life Estimation of Frankfurter Sausage Containing Dietary Fiber from Rice Bran Using Predictive Modeling (예측미생물을 이용한 미강식이섬유 함유 프랑크푸르터 소시지의 유통기한 설정)

  • Heo, Chan;Kim, Hyoun-Wook;Choi, Yun-Sang;Kim, Cheon-Jei;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.29 no.1
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    • pp.47-54
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    • 2009
  • Predictive modeling was applied to study the growth of microorganisms related to spoilage in frankfurter sausage containing various levels of dietary fiber (0, 1, 2, and 3%) from rice bran and to estimate its shelf-life. Using the Baranyi model, total viable cells, anaerobic and psychrotrophic bacteria were measured during 35 days of cold storage ($<4{\pm}1^{\circ}C$). The lag times (LT) demonstrated by control and treatment groups were 6.28, 623, 6.24, and 6.25 days, respectively. The growth rate of total viable cells in each group were 0.95, 0.91, 0.92, and 0.91 (Log CFU/g/day), respectively. The anaerobic and psychrotrophic bacteria had lower initial ($y_0$) and maximal bacterial counts ($y_{max}$) than total viable cells. Also, the anaerobic and psychrotrophic bacteria possessed lower growth rate and longer lag time than total viable cells. The estimated shelf-life of frankfurter containing rice bran fiber by the growth rate of total viable cells was 7.8, 7.9, 7.9, and 7.7 days, respectively. There were no significant differences in shelf-life as a function of fiber content. In other words, the addition of dietary fiber in sausage did not show the critically hazardous results in growth of microorganism. The 12 predictive models were then characterized by high $R^2$, and small RMSE. Furthermore, $B_f$ and $A_f$ values showed a very close relationship between the predictive and observed data.

Inorganic Phosphate Solubilization by Immobilized Pantoea agglomerans under in vitro Conditions (고정화된 Pantoea agglomerans에 의한 난용성 인산의 가용화)

  • Kim, Eun-Hee;Park, Sung-Ae;Park, Myoung-Su;Yang, Jin-chul;Madhaiyan, Munusamy;Seshadri, Sundaram;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.37 no.1
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    • pp.36-40
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    • 2004
  • It is now widely accepted that immobilized microbial cells can overcome some of the problems associated with microbial survival stability, efficacy, storage, transportation and ease of application in agricultural environments. Pantoea agglomerans, a phosphate solubilizing bacterium, was immobilized in alginate, agar and gelatin carriers. All the three immobilfized carriers with bacterial cells of P. agglomerans were compared for solubilization of tricalcium phosphate in pure liquid cultures. While alginate beads were tested for phosphate solubilization on alternate days up to five days, agar beads and gelatin cubes were subjected for one time phosphate solubilization analysis after seven days. Both alginate and agar immobilized cells of P. agglomerans exhibited higher efficiency in increasing the solubilizaliun of tricalcium phosphate than gelatin immobilized cells. The culture filtrate of alginate bead inoculation treatment registered a rapid increase in soluble phosphate concentration upon incubation. A corresponding decrease in the pH of the medium was also observed in all the treatments.