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Usefulness of Escherichia coli-expressed Recombinant VP6 Proteins of Group A Rotavirus in Serodiagosis of Rotavirus Infection (대장균에서 발현된 A군 로타바이러스 VP6 단백질을 이용한 로타바이러스 감염의 혈청학적 진단의 유용성)

  • Seo, Ji-Hyun;Kim, So-Young;Park, Ji-Sook;Lim, Jae-Young;Park, Chan-Hoo;Woo, Hyang-Ok;Youn, Hee-Shang;Kim, Won-Yong;Kang, Hyung-Lyun;Baik, Seung-Chul;Lee, Woo-Kon;Cho, Myung-Je;Rhee, Kwang-Ho
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.13 no.2
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    • pp.134-145
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    • 2010
  • Purpose: The serologic diagnosis of rotaviral infections is not commonly used in clinical practice, but is used in seroepidemiologic studies. In this study, the usefulness of Escherichia coli-expressed recombinant VP6 proteins of group A rotavirus in the serodiagnosis of rotavirus infections by ELISA was evaluated. Methods: The recombinant VP6 proteins of group A rotavirus expressed in E. coli Rosetta II strain were purified and identified. One hundred sera from 22 children (4 healthy neonates, 13 healthy children, and 5 immunocompromised children) who had serial sera samples prior to and after rotavirus infections were provided by the Gyeongsang National University Hospital, a member of the National Biobank of Korea. IgG, IgA, and IgM antibodies against rVP6 were analyzed by ELISA in all of the patients and Western blot analysis in 4 neonates. Results: ELISA tests using rVP6 proteins of group A rotavirus as antigen revealed that IgG, IgA, and IgM antibodies increased after rotaviral infections in most neonates and healthy children. IgG antibodies also increased after rotaviral infections in most immunocompromised children without an adequate increase in IgM or IgA antibodies. Western blot analysis in four neonates revealed very early IgM antibody responses, even in the sera with low optical densities in ELISA tests. Conclusion: Our study showed that ELISA using rVP6 as an antigen is a valid diagnostic tool for seroepidemiologic studies of rotavirus infections and Western blot analysis is a sensitive test in detecting IgG, IgA, and and IgM antibodies in patients with rotavirus infections.

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 transferring the effects of brand reputation and level of service satisfaction of an offline channel company when it is expanding to an online distribution channel (온라인 유통채널 확장시 오프라인 채널의 브랜드 명성, 서비스 만족도의 이전 효과에 관한 연구)

  • Hwang, Hee-Joong;Lee, Sun-Mi
    • Journal of Distribution Science
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    • v.9 no.2
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    • pp.31-36
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    • 2011
  • I conducted empirical analyses of what happens when an offline channel expands to an online channel and whether the pre-existing offline channel's competitive assets (e.g. brand reputation and level of service satisfaction) can be linked to online channel preference. I found that an offline channel's brand reputation and level of service satisfaction can have a direct influence on offline channel preference and a second-hand influence on online channel preference. Thus, if the competitiveness of the online channel is strong enough and its customers have a higher preference for the offline channel, they will be committed and loyal to the company. The resultant enhanced competitiveness of the offline channel will present opportunities for both present and future success. The main results are the following. First, the management of the distribution channel service quality is more important than that of the brand reputation. Customers' experiences of service and subjective evaluations are not important only as the leading factors in the long-term brand reputation management but also as influential factors in channel preference. SoThus, given that the service quality of the pre-existing channel is not the customers' main concern, a strategy of improving the level of service satisfaction aimed at present customers is more valuable than a wide brand positioning strategy aimed at general and new customers. Second, when an offline channel company establishes an internet shopping mall on an online channel, it is highly likely that the preference and subjective evaluation of the present customers will influence the online channel. This applies not only to the special case of an expansion from an offline intermediary channel to an online one, but also to an online channel acting as an expansion of the business model of a conventional manufacturing or service company: both cases are vertical integrations of marketing channels in an expansion of the distribution channel. My theory applies to a wide range of contexts. Third and finally, any business strategy can grasp the meaning of 'channel expansion. Fundamentally, it is an expansion of the sales activity channel and marketing activity. However, it is also a way of enhancing marketing and sales competitiveness through an expansion to an online or offline channel. The expansion of an offline company to an online channel could be seen not as improvement but as an innovation of the business process by which two goals are achieved with one technique. The former is expected to increase the sales of the offline company, and the latter is also expected to increase sales while also contributing to cost reduction.

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Enzymatic Hydrolysis of Yellowfin Sole Skin Gelatin in a Continuous Hollow Fiber Membrane Reactor (연속식 중공사막 반응기를 이용한 각시가자미피 젤라틴의 가수분해)

  • KIM Se-Kwon;BYUN Hee-Guk;KANG Tae-Jung;SONG Dae-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.26 no.2
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    • pp.120-132
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    • 1993
  • A continuous hollow fiber membrane reactor(CHFMR) was developed and optimized for the production of yellowfin sole(Limanda aspera) skin gelatin hydrolysates using trypsin. The results were summerized as follows: The $K_m$ value of the CHFMR was 2.4 times higher than that of the batch reactor, indicating reduced enzyme affinity for the substrate. The $K_2$ value of the CHFMR was 8.5 times lower than that of the batch process, showing a significant reduction in trypsin activity in the CHFMR. The optimum operating conditions for the CHFMR process were $55^{\circ}C$, pH 9.0, flux 7.79 ml/min, residence time 77min, and trypsin to substrate ratio, 0.01(w/w) After operating for 60min under the above conditions, $79\%$ of the total amount of initial gelatin was hydrolysed. Enzyme leakage was observed through the 10,000 MWCO membrane after the 20min of reactor operation, while none occurred after 5hr. Total enzyme leakage was about $12.95\%$ at $55^{\circ}C$ for 5hrs. However, there was no apparent correlation between enzyme leakage and substrate hydrolysis. The membrane has a significant effect on trypsin activity loss for 60min of the CHFMR operation. The CHFMR operating with the membrane lost $34\%$ of the initial activity versus a $23\%$ loss of activity after 3hr in the continuous reactor lacking the hollow fiber membrane. The measurement of fouling property showed that relative flux reduction was $91\%$ and flux recover rate was $92\%$ at $10\%$ substrate solution. The productivity(378.85mg product/mg enzyme) of the CHFMR was more than 4 times higher than that of the batch reactor at $55^{\circ}C$.

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고무의 가황(加黃) 및 열전도론(熱傳導論) (3(三))

  • Heo, Dong-Seop;Gwon, Dong-Yong
    • Elastomers and Composites
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    • v.10 no.2
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    • pp.136-156
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    • 1975
  • 고무는 불량열전도체(不良熱傳導體)이며 두께가 두꺼우면 내부(內部)가 적정온도수준(適正溫度水準)에 이르기 전까지 가황시간(加黃時間)이 길어진다. 가황온도(加黃溫度)가 상승(上昇)할수록 가황물(加黃物)의 물성(物性)은 열화(劣化)되는 경향(傾向이) 있다. 천연(天然)고무든지 합성(合成)고무든지 간(間)에 과가황(過加黃)에 대(對)한 저항성(抵抗性)이 나쁘므로 특(特)히 고온가황(高溫加黃)에 대(對)해 민감(敏感)하다. 이것은 고온(高溫)에서 단시간(短時間) 가황(加黃)일수록 가속(加速)된다. 평탄가황배합물(平坦加黃配合物)의 경우에서 보더라도 내부(內部)가 적절(適切)히 가황(加黃)되기도 전(前)에 외부(外部)는 과가황(過加黃)이 되는 수가 있다. 근래(近來) 발간(發刊)된 문헌(文獻)에서도 이러한 내용(內容)이 잘 설명(說明)이 되어 있는데 다른 각도(角度)에서 고찰(考察)해 볼것 같으면 정체시간(停滯時間)이 비교적(比較的) 길지 않는 한(限) 가황시간(加黃時間)은 정체시간(停滯時間)과 sheet 가황시간(加黃時間)과의 합(合)이라고 말할 수 있겠다. 예(例)를 들어 설명(說明)하자면 $130^{\circ}C(266^{\circ}F)$에서 정체시간(停滯時間)이 10분(分)이고 sheet 가황시간(加黃時間)이 20분(分)인 제품(製品)은 이 온도(溫度)에서 30분간(分間) 가황(加黃)해야 된다는 것이다. 온도계수(溫度係數)를 2라고 가정(假定)할 경우 $140^{\circ}C(284^{\circ}F)$에서의 가황시간(加黃時間)은 $30\times\frac{1}{2}=15$분(分)이 아니라 $20\times\frac{1}{2}+10=20$분(分)이 된다. 크기가 큰 제품(製品)은 보통(普通) 다음에 있는 여러 방법(方法)들 가운데 한 가지 또는 여러가지를 조합(組合)하여 가황(加黃)시킨다. a) 크기가 작은 것에 대한 것 보다 낮은 온도(溫度)에서 가황(加黃)한다. b) 침투가황-제품(浸透加黃-製品)을 가압하(加壓下)에 두고서 외부가황(外部加黃)은 단속(斷續)시키고 열(熱)이 중심(中心)으로 침투(浸透)하게 한다. c) 단계가황(段階加黃)-처음에는 저온(低溫)에서 시작(始作)하여 일정간격(一定間隔)을 두고 점차(漸次) 온도(溫度)를 상승(上昇)시켜 최종적(最終的)으로 가황온도(加黃溫度)까지 올린다. d) 가능(可能)하다면 metal base나 금형(金型)에서 고무를 증기가황(蒸氣加黃)시킬 경우에 있어서 속이 빈 축(軸)을 사용하여 내부(內部)로 부터 가열(加熱)하면 가황시간(加黃時間)이 단축(短縮)된다. e) 냉각중(冷却中)의 후가황(後加黃)-이것은 가열장치(加熱裝置)에서 끄집어낸 후 제품(製品)의 외부(外部)를 냉각(冷却)시키는 방법(方法)이다. 가열(加熱)된 제품(製品)이 쌓여 있거나 적절(適切)하게 냉각(冷却)되지 않을 때 가황(加黃)이 추가적(追加的)으로 되거나 과가황(過加黃)이 될 우려가 있는 제조공정(製造工程)에서는 흔히들 이 방법(方法)을 무시(無視)하고 있다. 여기서 강조(强調)해 두어야 할 것은 항상 제품(製品)의 외부(外部)를 완전(完全)히 가황(加黃)시킬 필요(必要)는 없다는 것이다. 다공성(多孔性)이나 기포생성(氣泡生成)을 조장(助長)하는 불량가황상태(不良加黃狀態)와 표면(表面)에서의 과가황상태간(過加黃狀態間)의 균형(均衡)을 취(取)해 줘야 하는데 물론(勿論) 이때는 가황시간(加黃時間)을 단축(短縮)시켜야 한다는 경제적(經濟的)인 측면(側面)도 아울러 고려(考慮)해야 한다. 이것은 고무기술자(技術者)가 당면(當面)해야할 과제(課題)에 속(屬)하며 바람직 한것은 본장(本章)의 내용(內容)이 여러 상황하(狀況下)에서 당면(當面)한 문제(問題)에 대(對)해 어떻게 대처(對處)해 야 할지를 모르는 여러 기술자(技術者)들에게 도움이 되었으면 하는 것이다.

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Antioxidative and Antiaging Effects of Jeju Native Plant Extracts(II) (제주 자생 식물 추출물의 항산화 및 항노화에 관한 연구(II))

  • Park, Soo-Nam;Kim, Jin-Young;Yang, Hee-Jung;Lee, Keun-Ha;Jeon, So-Mi;Ahn, You-Jin;Won, Bo-Ryoung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.3
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    • pp.165-173
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    • 2007
  • In the previous study, we reported the antioxidative and cellular protective effects of Jeju native plant extracts. In this study, we investigated the anti-oxidative, anti-wrinkle and whitening effects of new 37 plant extracts collected from self-growing plants in Jeju island. Their anti-oxidant activities were measured by the 1,1-diphenyl-2-picrylhydrazyl(DPPH) radical scavenging assay and reactive oxygen species(ROS) scavenging assay in $Fe^{3+}-EDTA/H_2O_2$ system. The cytoprotective properties of 37 plant extracts were assessed in the rose-bengal sensitized photohemolysis of human erythrocytes. The inhibitory effect of 37 plant extracts on tyrosinase were investigated to assess their whitening efficacy. Finally, their anti-elastase activities were measured to predict the anti-wrinkle efficacy in the human skin. The results showed that the extracts of Myrica rubra stem bark and Securinega suffruticosa have the free radical scavenging activity($FSC_{50}:\;5,\;8{\mu}g/mL$, respectively), and the extracts of Quercus acutissima leaf and Securinega suffruticosa stem bark have the prominent ROS scavenging activity($OSC_{50}:\;0.009{\mu}g/mL$). Photohemolysis of erythrocytes in the presence of rose-bengal as a sensitizer was inhibited by the extracts of Securinega suffruticosa stem bark and Salix koreensis stem(${\tau}_{50}$, 895 min, 640 min at 50 ${\mu}g/mL$, respectively. Myrica rubra stem bark extract(77.8% at 200 ${\mu}g/mL$) and Salix koreensis stem extract(76.2% at 200 ${\mu}g/mL$) also have the inhibitory effect on tyrosinase and elastase activities, respectively. These results indicated that the stem park of Myrica rubra, Securinega suffruticosa, and Camellia japonica, the stem of Salix koreensis, and the leaf of Quercus aqutissima and Camellia japonica could have e benefitial effects when they are added as ingredients in cosmetics.

Formation of the $CoSi_{2}$ using Co/Zr Bilayer on the Amorphous and the Single Crystalline Si Substrates (단결정과 비정질 Si 기판에서 Co/Zr 이중층을 이용한 $CoSi_{2}$ 형성)

  • Kim, Dong-Wook;Jeon, Hyeong-Tag
    • Korean Journal of Materials Research
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    • v.8 no.7
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    • pp.621-627
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    • 1998
  • The formation of Co-silicide between Co/Zr bilayer on the amorphous and crystalline Si substrates has been investigated. The films of Zr(50$\AA$) and Co(l50$\AA$) were deposited with e-beam evaporation system and were heattreated with the rapid thermal annealing system at the temperatures between 50$0^{\circ}C$ and 80$0^{\circ}C$ with 10$0^{\circ}C$ increments for 30 seconds. The phase identification of Co-silicide was carried out by XRD and the chemical analysis was examined by AES and RBS. The interface morphologies of Co/Zr bilayer films were investigated by cross sectional TEM and HRTEM. $CoSi_2$ was formed epitaxially on the crystalline Si substrate above $700^{\circ}C$ while polycrystalline $CoSi_2$ was grown on the amorphous Si substrate. The formation temperature of Co-silicide on the amorphous Si substrate was about 100 C lower than that on the crystalline Si. The COzSi phase was not identified on the both Si substrates. The formation temperature of first phase of Co-silicide on ColZr bilayer was higher than that on Co mono layer. CoSizlayer formed on the amorphous Si substrate exhibits better uniformity compared to the CoSiz formed on the crystalline substrate. The sheet resistance of CoSiz layer on crystalline Si was lower than that on the amorphous Si at high temperatures.tures.

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Crystallization and Magnetic Properties of Iron Doped La-Ba-Mn-O (Fe이 치환된 LaBaMnO계 산화물의 중성자 회절 및 Messbauer분광학연구)

  • Choi, Kang-Ryong;Kim, Sam-Jin;Shim, In-Bo;Kim, Chul-Sung
    • Journal of the Korean Magnetics Society
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    • v.14 no.1
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    • pp.38-44
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    • 2004
  • The iron doped colossal magnetoresistance materials with La-Ba-Mn-O perovskites structure have been synthesized by chemical reaction of sol-gel methods. Their crystallographic and magnetic properties have been studied with x-ray diffraction, VSM, RBS, Mossbauer spectroscopy, and magnetoresistance measurements. The crystal structure of the La$\_$0.67/Ba$\_$0.33/Mn$\_$0.99/Fe$\_$0.01/ $O_3$ at room temperature was determined to be orthorhombic of Pnma. The lattice parameters a$\_$0/ and c$\_$0/ increased gradually, but b$\_$0/ deceased with increase of iron substitution. The magnetization and coercivity deceased, also the Curie temperature decreased from 360 K as x increased from 0.00 to 0.05. Magnetoresistence measurements were carried out, and the maximum MR ($\Delta$$\rho$/$\rho$(0)) was observed at 281 K, about 9.5 % in 10 kOe. The temperature of maximum resistance (R$\_$MAX/) decreased with increasing substitution of Fe ions and a semiconductor-metal transition temperature (T$\_$SC-M/) decreased too. This phenomena show that ferromagnetic transition temperature decreased by substituting Fe for Mn ions, it decreases double exchange interaction. This result accords with magnetic structure of neutron diffraction. Mossbauer spectra of La$\_$0.67/Ba$\_$0.33/Mn$\_$0.99/Fe$\_$0.01/ $O_3$were taken at various temperatures ranging from 15 to 350 K. With lowering temperature of the sample, two magnetic phases were increased and finally it showed the two sharp sextets of spectra at 15 K. The isomer shift at all temperature range is about 0.3 mm/s relative to Fe metal, which means that both Fe ions are Fe$\^$3+/ states.Fe$\^$3+/ states.

Construction and characterization of heterozygous diploid Escherichia coli (2배체 대장균의 제조와 그 특성)

  • Jung, Hyeim;Lim, Dongbin
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.406-414
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    • 2016
  • Among 6 leu codons, CUG is the most frequently used codon in E. coli. It is recognized by leu-tRNA(CAG) encoded by four genes scattered on two chromosomal loci (leuT and leuPQV ). In the process of constructing a strain with no functional leu-tRNA (CAG) gene on chromosome, we made two mutant strains separately, one on leuPQV locus (${\Delta}leuPQV$), and the other on leuT locus [$leuT^*$(GAG)], where the anticodon of leuT was changed from CAG to GAG, thereby altering its recognition codon from CUG to CUC. We attempted to combine these two mutations by transduction using $leuT^*$(GAG) strain as a donor and ${\Delta}leuPQV$ strain as a recipient. Large and small colonies appeared from this transduction. From PCR and DNA sequencing, large colony was confirmed to be the reciprocal recombinant as expected, but the small colonies contained both mutant $leuT^*$(GAG) and wild type leuT (CAG) genes in the cell. This heterozygous diploid strain did not show any unusual morphology under microscopic observation, but, interestingly, it showed a linear growth curve in rich medium with much slower growth rate than wild type cell. It always formed homogenous small colonies in the selection medium, but, when there was no selection, it readily segregated into $leuT^*$(GAG) and leuT (CAG). From these observations, we suggested that the strain with both $leuT^*$(GAG) and leuT (CAG) genes was not a partial diploid (merodiploid), but a full diploid cell having two different chromosomes. We proposed a model explaining how such a heterozygous diploid cell was formed and how and why its growth showed a linear growth curve.

Quality and Functional Components of Commercial Chungkukjang Powders (시판 청국장 분말제품의 품질 및 기능성분)

  • Lee, Hyo-Jin;Cho, Sang-A;Shin, Jin-Gi;Kim, Jeong-Sang;Jeong, Yong-Jin;Moon, Kwong-Duck;Kwon, Joong-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.1
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    • pp.65-71
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    • 2007
  • Five different commercial chungkukjang powders (A$\sim$E) were compared based on their physicochemical, organoleptic and functional properties. The proximate composition of the five samples ranged from 6.07 to 8.54% in moisture, 15.31 to 27.07% in crude protein, 20.19 to 24.75% in crude lipid, 34.84 to 52.41% in carbohydrate, and 3.69 to 5.26% in ash. The pH of the samples ranged from 5.58 to 6.11, and Hunter's colors showed 70.01$\sim$77.22 for L value, 0.91$\sim$4.64 for a value and 23.72$\sim$31.00 for b value depending on the product. The microbial counts were 8.16$\sim$g.60 log CFU/g for aerobic bacteria, $\sim$4.16 log CFU/g for yeasts & molds, and 1.07$\sim$3.88 log CFU/g for coliforms. The contents of reducing sugars and amino-N were 1.89$\sim$2.41% and 2.83$\sim$7.31%, respectively. Free amino acids were mainly composed of glutamic acid, valine, leucine, phenylalanine, and lysine. The amounts of total phenolics and total flavonoids were 108$\sim$302 mg% and 2.73$\sim$9.41, respectively, with some variations in the products. However, the isoflavone contents, which were composed of genistein (63.26$\sim$217.16${\mu}g/g$), daidzein (58.24$\sim$166.65${\mu}g/g$), genistin (2.66$\sim$55.68${\mu}g/g$), and glycitein (12.26$\sim$17.82${\mu}g/g$), were apparently different per product. The sensory scores for color, smell, taste and overall preference for the five chungkukjang products, which were evaluated by panels in their 20's and 30's using 7-point scoring test, ranged from 3.20 to 4.05.