• Title/Summary/Keyword: viable cells

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Conditions for Stable light Production of Recombinant Escherichia coli Containing Lux Operon and Sensitivity to Toxic Chemicals (Lux operon을 함유한 유전자 재조합 Escherichia coli의 발광 안정화 조건 및 독성물질에 대한 민감성)

  • 배희경;이상민;정윤철;송방호;신평균
    • KSBB Journal
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    • v.17 no.6
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    • pp.571-576
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    • 2002
  • Recombinant E. coli DH5 ${\alpha}$/pSB311 was made by cloning the genes encoding bacterial luciferase and aldehyde substrate proteins from Photohabdus luminescense, to complement defects of Lumistox, which is normally used in bioassays to monitor toxic substances in water environmental systems. The conditions for stable light production by the recombinant strains were investigated with respect to cell growth stage, cell number, and buffer conditions. The optimum growth stage was a middle-exponential stage with an OD$_{660nm}$ value of 0.6-0.7. ADout 10$^{6}$-10$^{7}$ cells per test tube was optimum for stable light emission. The effect of buffer was not significant if an optimum viable cell number was maintained. The bioluminescence of the recombinant E. coli harboring the lux operon of Photohabdus luminescense was not affected by temperature, while the bioluminescence of Lumistox was temperature sensitive. The recombinant E. coli was more sensitive to heavy metals (Cd, Cu, Hg, Zn) than Lumistox, because it does not require high concentrations of NaCl in the buffer.

Quality characteristics of Doenjang prepared with sweet potato (고구마를 이용한 된장의 품질 특성)

  • Cha, Su-Jin;Park, Sao-Ra;Kim, Dong-Han
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.221-229
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    • 2017
  • The effect of sweet potato on the quality of Doenjang was investigated during fermentation. Viable cells of yeast decreased gradually after 4 weeks of fermentation, but those of aerobic bacteria increased in the late stage. Amylase activity of Doenjang was higher in the late stage of fermentation, while neutral protease maintained high activity during fermentation. Hunter L and b values of Doenjang decreased gradually during fermentation, while a value was increased. The pH of Doenjang decreased gradually until 10 weeks of fermentation, and the titratable acidity was low in the sweet potato added groups. The acid value was low in the Shinyulmi sweet potato added Doenjang. Water activity and oxidation-reduction potential of Doenjang decreased during fermentation. Reducing sugar of Doenjang decreased in the middle stage of fermentation, and it was low in sweet potato added groups. The alcohol content of Doenjang decreased after 2 weeks of fermentation. Amino and ammonia-type nitrogen of Doenjang increased during fermentation and reached the maximum after 10 and 12 weeks of fermentation, respectively. After 12 weeks fermentation, 8% of Shinyulmi sweet potato added Doenjang was more favorable taste, flavor and overall acceptability (p<0.05) than the control or the Yeonwhangmi sweet potato added groups.

Characteristics of yeast with low temperature adaptation for Yakju brewed (약주 제조를 위한 저온 적응성 효모의 특성)

  • Seo, Dong-Jun;Yeo, Soo-Hwan;Mun, Ji-Young;Jung, Woo-Jin;Cho, Yong Sik;Baek, Seong Yeol
    • Food Science and Preservation
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    • v.22 no.6
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    • pp.908-914
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    • 2015
  • The objectives of this study were to isolate and characterize low temperature adaptation yeast and to obtain suitable yeasts strains for manufacturing Yakju. In this study, we isolated 482 wild yeasts from fermented foods. Out of these, 5 yeast strains were selected based on increased growth at low temperature ($15^{\circ}C$) and high ${\beta}$-glucosidase activity. To screen the aromatic level of isolates, media containing cerulenin and 5,5,5-trifluor-DL-leucine (TFL) were used. Y297 strain demonstrated tolerance against TFL and produced more than 13% alcohol. Y297 strain was identified a Saccharomyces cerevisiae based on the 26S rDNA gene sequences. Maximum cell growth was observed after 19 hr and 38 hr of incubation at $25^{\circ}C$ and $15^{\circ}C$, respectively. The exponential phase was followed by a lengthy stationary phase, at $15^{\circ}C$, when the cells remained high viable. Y297 strain demonstrated tolerance against alcohol (10%), glucose (60%) and salt(NaCl, 8%). ${\beta}$-glucosidase and esterase activity in Y297 were higher than those of controls at $15^{\circ}C$. Overall, these results indicated that using wild yeast strain, isolated from fermented food, affects the chemical characteristics of the brewed Yakju.

Microbiological Quality of Myungran Jeotkal and Its Ingredients and Improvement of Shelf-stability by Gamma Irradiation (명란젓갈 및 부재료의 미생물 오염도 및 감마선 조사를 이용한 유통안정성 향상)

  • Kim, Bin-Na;Jang, Ae-Ra;Song, Hyun-Pa;Kim, Yun-Ji;Ko, Byung-Ho;Jo, Cheorun
    • Food Science and Preservation
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    • v.15 no.4
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    • pp.606-611
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    • 2008
  • Myungran Jeotkal, Korean fermented seafood, and its ingredients(hot red pepper powder, ginger, garlic, and seasoning mix) were irradiated with 0, 0.5, 1.0, 2.0, and 5.0 kGy of gamma rays and stored at 4C for 4 weeks to determine changes in microbiological and sensory characteristics. Water activities of Myungran Jeotkal, hot red pepper powder, ginger, garlic, and seasoning mix were 0.89 0.56, 0.98, 0.99, and 0.07, respectively. Myungran Jeotkal was observed to be initially contaminated. Total aerobic bacteria, yeast and mold, and coliform levels were 6.7, 4.3, and 3.6 log CFU/g, respectively. Irradiation at 2 kGy afforded approximately a 4 log reduction in total aerobic bacteria, and a 3 log drop in both yeast and mold levels and coliform bacteria(P<0.05). No viable microbial cells were detected in Myungran Jeotkal after 5 kGy of irradiation(at a detection limit of 101 CFU/g). The total aerobic bacterial level in red pepper powder was 6.3 log CFU/g and this component, of the tested ingredients, contributed most to the microbial contamination of Myungran Jeotkal. The initial count of total aerobic bacteria, 6.3 log CFU/g, was significantly reduced to 4.5 log CFU/g after irradiation(P<0.05). Sensory evaluation showed that gamma irradiation of up to 5.0 kGy did not adversely affect overall acceptability of Myungran Jeotkal or its ingredients during cold storage. Therefore, gamma irradiationwas effective to extend the shelf-life of Myungran Jeotkal.

Prevalence of virulence-associated genes and antimicrobial resistance of Campylobacter jejuni from ducks in Gyeongnam Province, Korea

  • Yang, Jung-Wong;Kim, Sang-Hyun;Lee, Woo-Won;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.37 no.2
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    • pp.85-96
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    • 2014
  • Total 99 strains of Campylobacter spp. were isolated from 117 cases of duck's fecal samples. Among 99 strains of Campylobacter spp. isolates, 93 strains (93.9%) were C. jejuni and 6 strains (6.1%) were C. coli. Prevalence of virulence and GBS associated genes of 72 C. jejuni isolates was determined by m-PCR. Among the 10 kinds of virulence associated genes, cadF, dnaJ, flaA and ceuE genes were detected in all of C. jejuni isolates from ducks, racR, pldA, iamA, ciaB, virB11 and docC genes were 87.5%, 84.7%, 77.8%, 48.6%, 13.9% and 11.1%, respectively. Antimicrobial susceptibility test was performed on 72 C. jejuni isolates. The rate of resistance were 62.5% for oxytetracycline, 55.6% for kanamycin, 54.2% for enrofloxacin, 50% for ciprofloxacin, 37.5% for tetracycline and nalidixic acid, 18.1% for ampicillin, 15.3% for streptomycin, and 6.9% for ofloxacin. All isolates were susceptible to erythromycin. The adherence (intracellular and extracellular bacteria) abilities of the 20 isolates to INT-407 cells were between $4.21{\pm}1.27{\times}10^4$ CFU/well and $1.053{\pm}0.451{\times}10^6$ CFU/well from the isolates of cj-55 and cj-52, respectively, and that can be expressed as 0.1033% to 5.2655% to the infecting inoculum. The invasion (intracellular bacteria) abilities of the 20 isolates to INT-407 were between $1.00{\pm}1.73{\times}10^3$ CFU/well and $8.47{\pm}5.16{\times}10^4$ CFU/well from the isolates of cj-13 and cj-47, respectively, and that can be expressed as 0.0050% to 0.4235% to the infecting inoculums. The average CFU/well of 20 campylobacters isolated from ducks for adherence to and invasion were $2.646{\pm}2.886{\times}10^5$ and $3.03{\pm}2.7{\times}10^4$ respectively, and that was $1.3230{\pm}1.2139%$ and $0.1516{\pm}0.1343%$ of the starting viable inoculum. There was considerable correlation ($R^2$=0.627) between the adherence and invasion ability of C. jejuni isolates for INT-407 cell.

Characteristics of White Soybean Chungkookjang Fermented by Bacillus subtilis D7 (Bacillus subtilis D7에 의하여 발효된 백태 청국장의 특성)

  • Lee, Na-Ri;Park, Sung-Bo;Lee, Sang-Mee;Go, Tae-Hun;Hwang, Dae-Youn;Kim, Dong-Seob;Jeong, Seong-Yun;Son, Hong-Joo
    • Journal of Life Science
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    • v.23 no.4
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    • pp.529-536
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    • 2013
  • This study was carried out to investigate the characteristics of white soybean Chungkookjang fermented by Bacillus subtilis D7. The highest germination rate was obtained under $25^{\circ}C$ when water was supplied for 4 days at intervals of 2 hr. The total isoflavone content was 971.3 ${\mu}g/g$ before germination and 1023.8 ${\mu}g/g$ after germination. The amino type- and ammonia type-nitrogen contents of Chungkookjang were proportional to the fermentation time. The pH values of all Chungkookjang soybeans increased up to pH 7.8-8.0 during fermentation. The number of viable cells in all Chungkookjang soybeans increased significantly up to 24 hr. The protease activity of all Chungkookjang soybeans increased up to 30-36 hr. The ${\alpha}$- and ${\beta}$-amylase activities of Chungkookjang fermented by B. subtilis D7 were enhanced with increasing fermentation time. Analysis of the effect of Chungkookjang intake on the liver function of mouse showed that the alkaline phosphatase (ALP) activity and the superoxide dismutase (SOD) activity in the Chungkookjang diet group were markedly higher than those in the control group. The asparatate aminotransferase (AST) activity in the germinated soybean Chungkookjang diet group was higher than that in the nongerminated soybean Chungkookjang diet group. Therefore, Chungkookjang soybeans fermented with B. subtilis D7 can be expected to have an increased content of functional components and improved quality characteristics.

Real-Time Monitoring of Mitochondrial ATP Synthesis and Hydrolysis by Surface Infrared Spectroscopy

  • Yamaguchi, Ryo-Taro;Hirano-Iwata, Ayumi;Aonuma, Yuki;Yoshimura, Yuya;Shinohara, Yasuo;Kimura, Yasuo;Niwano, Michio
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.108-109
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    • 2013
  • Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

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Microbiological and Mutagenical Safety Evaluation of Gamma Irradiated Ready-to-Eat Foods of Animal Origin (즉석식품 제조를 위한 육가공제품의 감마선 조사에 따른 미생물 및 유전독성학적 안전성 평가)

  • Lee Na-Young;Jo Cheorun;Kang Ho-Jin;Hong Sang-Pill;Kim Young-Ho;Lee Kyong-Haeng;Byun Myung-Woo
    • Food Science of Animal Resources
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    • v.25 no.1
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    • pp.13-19
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    • 2005
  • The radio-sensitivity of pathogens and the effect of irradiation on microbiologican safety and mutagenicity of meat products such as seasoned and cooked beef and ham were investigated. Samples were radiation-sterilized and inoculated at 10/sup 7/ cfu/g with each of the four pathogens including Salmonella typhimurium, Escherichia coli, Staphylcoccus aureus, and Listeria ivanovii. No viable cells of pathogens were observed in the sample irradiated with 3 kGy. The D/sub 10/ value of inoculated pathogens in seasoned and cooked beef and ham were 0.24∼0.48 and 0.39∼0.45, respectively. Results of Ames test performed with non-irradiated and irradiated seasoned and cooked beef and ham were both negative at the level of 625, 1,250, 2,500, 50,000, and! 10,000 ㎍ sample/plate, respectively. Results indicate that low dose (2∼3 kGy) irradiation is effective to ensure safety for seasoned and cooked beef and ham with toxicological wholesomeness.

Desalination of Traditional Soy Sauce Using Electrodialysis (전기투석에 의한 재래식 간장의 저염화)

  • Chung, Jae-Ho;Mok, Chul-Kyoon;Lim, Sang-Bin;Woo, Gun-Jo;Baek, Hyung-Hee;Park, Young-Seo
    • Korean Journal of Food Science and Technology
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    • v.34 no.5
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    • pp.811-817
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    • 2002
  • Korean traditional soy sauce was desalted using electrodialyzer, and their physicochemical properties were analyzed. The salt content of soy sauce significantly decreased from initial 18.9 (w/v) to 0.47% (w/v) and the volume also decreased to 330 mL when 1,000 mL of soy sauce was desalted for 450 min at a current of 2 A. During the desalting process, the numbers of viable cells of total bacteria, yeasts, and molds increased due to the concentration effect of the electrodialysis. Turbidity increased from 3.1 to 8.5, and total nitrogen, total sugar, and reducing sugar contents also increased. The pH, and amino nitrogen and volatile base nitrogen contents slightly decreased, with ammonia content significantly decreasing from 19.4 to $1.3\;{\mu}mol/mL$. Free amino acids content increased twofold after electrodialysis. Gas chromatograms of soy sauce showed that profiles of the flavoring compounds did not change when electrodialyzed for 360 min, but most were removed after 540 min.

Effect of Condiments on the Microflora, Enzyme Activities and Taste Components of Traditional Kochujang during Fermentation (양념류를 첨가한 전통고추장의 숙성중 미생물, 효소활성 및 맛 성분의 변화)

  • Kim, Dong-Han
    • Korean Journal of Food Science and Technology
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    • v.33 no.2
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    • pp.264-270
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    • 2001
  • In order to improve the quality of traditional kochujang, condiments like garlic and onion were added to kochujang and their effect on microbial characteristics, enzyme activities and taste components were investigated during fermentation. Viable cells of yeasts in the kochujang decreased with the increasing ratio of garlic. However, aerobic and anaerobic bacterial counts did not show any remarkable differences in the garlic or onion added kochujang, they decreased slowly after $6{\sim}10$ weeks of fermentation. The activity of liquefying amylase in kochujang decreased slowly during $2{\sim}10$ weeks of fermentation, but that of saccharifying amylase increased remarkably at $14{\sim}18$ weeks. Amylase activities increased at the late period of fermentation in garlic or onion added kochujang. Protease activities of kochujang were strong in the middle of fermentation, but they decreased by addition of garlic or onion. The major free sugars in kochujang were glucose and fructose, and their contents increased as the ratio of garlic increased. The major organic acids in kochujang were succinic, malic and oxalic acid, and they also decreased by addition of garlic and onion. The contents of total free amino acids and amino nitrogen were the highest in 2% garlic added kochujang. The major free amino acid were glutamic acid, aspartic acid, proline, alanine, leucine, isoleucine and arginine. Serine, glutamic acid, lysine and arginine contents in increased by adding garlic or onion.

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