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Production of Intracellular Invertase from Alkalophilic and Thermophilic Bacillus sp. TA-11 in the Recombinant E. coli (재조합 대장균에서 호알칼리성,고온성 Bacillus sp. TA-11의 세포내 Invertase의 생산)

  • Yi, Sung-Hun;Lee, Dae-Hyung;No, Jae-Duck;Lee, Jae-Won;Lee, Jong-Soo
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.318-322
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    • 2006
  • The intracellular invertase gene of alkalophilic and thermophilic Bacillus sp. TA-11 which was isolated from compost was cloned and expressed in E. coil HB101 using pUC19 as a vector. The invertase of the recombinant E. coli (pYC 17) was maximally produced when it was incubated at 37$^{\circ}C$ for 9 h in a SY medium containing 0.25% sucrose, 0.5% yeast extract, 0.1% each of $K_2HPO_4$ and $KH_2PO_4$, with an initial pH of 8.0. The final enzyme activity under the above condition was 47.7 U per ml of cell-free extract.

Molecular Cloning of a Gene Cluster for Phenanthrene Degradation from Pseudomonas sp. Strain DJ77 and Its Expression in Escherichia coli (Pseudomonas sp. strain DJ77로 부터 phenanthrene 분해 유전자군의 클로닝과 대장균에서의 발현)

  • 김영창;윤길상;신명수;김흥식;박미선;박희진
    • Korean Journal of Microbiology
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    • v.30 no.1
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    • pp.1-7
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    • 1992
  • We cloned a gene cluster encoding phenanthrene-degrading enzymes on a 6.8-kb Xhol fragment from the Pseudomonas sp. DJ77 chromosomal DNA into the vector pBLUESCRIPT SIC(+). The resultant clone, containing the recombinant plilsmid pHENX7, was able to convert 3-methylcatechol to a yellow mela-cleavage compound. Since the pHENX7R in which the DNA insert was cloned in the opposite orientation lacked extradiol dioxygenase activity. the direction of transcription was established. Four polypeptides, PhnC (24 kDa). PhnD (31 kDa), PhnE (34 kDa). and PhnF (15 kDa), were identified in E coli JM101 transformed with several pHENX7-derived plasmids. The locations and extents of ~ndividual genes were determined by subcloning. The gene order was phnC-phnD-phnE-phnF-phnG, and phnC, phnD, phnE, and phnG genes encoded glutathione S-transferase, mrta-cleavage compound hydrolase, extradiol dioxygenase, mera-cleavage compound dehydrogenase, respectively.

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Increment of Germanium Contents in Angelica keiskei Koidz. and Panax ginseng G.A. Meyer by In Vitro Propagation (명일엽(明日葉)(신선초(神仙草)) 및 인삼(人蔘)의 기내배양(器內培養)을 통한 Germanium 함량(含量) 증대(增大))

  • Lee, Man-Sang;Lee, Joong-Ho;Kwon, Tae-Oh;Namkoong, Seung-Bak
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.3
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    • pp.251-258
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    • 1995
  • This study was carried out to find optimum concentration of germanium compounds and pH of medium on the induction and growth of callus from A. keiskei and P. ginseng and to intend to increase Ge. absorption by calli while those calli were subculturing on MS medium. Callus from a. keiskei was rarely induced under light condition. Under dark condition, callus in­duction from A. keiskei was good up to 5ppm, retarded at 50ppm of $GeO_2$, or C. E. Ge. O., and rarely done at 100 ppm of $GeO_2$ but was somewhat well at 100 ppm of C. E. Ge. O. The induction and growth of callus was good in order of pH 5. 7 > pH 5. 4 > pH 6. 0 Under light condition, the growth of callus induced from P. ginseng was poor at $1{\sim}10\;ppm$ of $GeO_2$, or C. E. Ge. O., but shooting from callus occurred frequently. Under dark condtion, the growth of callus from A. keiskei was good up to 5 ppm of $GeO_2$, or C. E. Ge. O. and was rarely done at 50 ppm of $GeO_2$, but was somewhat well even at 100 ppm of C. E. Ge. O. Shooting from callus occurred frequently in a. keiskei, especially at pH 5.7. The growth of callus from P. ginseng was poor at 10 ppm of $GeO_2$, or 50 ppm of C. E. Ge. O. Under dark condition, the amount of Ge absorption by callus induced from A. keiskei was much high­er than that from P. ginseng. The amount of Ge. absorption by callus treated with $GeO_2$, was higher than that treated with C. E. Ge. O.

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Effects of Ensiling Period and Bacterial Inoculants on Chemical Compositions and Fermentation Characteristics of Rye Silage

  • Lee, Seong Shin;Joo, Young Ho;Choi, Jeong Seok;Jeong, Seung Min;Paradhipta, Dimas Hand Vidya;Noh, Hyeon Tak;Han, Ouk Kyu;Kim, Sam Churl
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.41 no.4
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    • pp.259-266
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    • 2021
  • The present study was aimed to estimate the effect of ensiling period and bacterial inoculants on chemical compositions and fermentation characteristics on rye silage harvested at delayed stage. Rye (Secale cereale L.) was harvested after 20 days of heading stage (29.4% dry matter, DM). The harvested rye forage was applied with different inoculants following: applications of distilled water (CON), Lactobacillus brevis (LBB), Leuconostoc holzapfelii (LCH), or mixture of LBB and LCH at 1:1 ratio (MIX). Each forage was ensiled into 20 L mini bucket silo (5 kg) for 50 (E50D) and 100 (E100D) days in triplicates. The E50D silages had higher in vitro digestibilities of DM (IVDMD, p<0.001) and neutral detergent fiber (IVNDFD, p=0.013), and lactate (p=0.009), and acetate (p=0.011) than those of E100D, but lower pH, lactic acid bacteria (LAB), and yeast. By inoculant application, LCH had highest IVDMD and IVNDFD (p<0.05), while MIX had highest lactate and lowest pH (p<0.05). The CON and LCH in E50D had highest LAB and yeast (p<0.05), whereas LBB in E100D had lowest (p<0.05). Therefore, this study concluded that LCH application improved the nutrient digesbility (IVDMD and IVNDFD) of lignified rye silage, and longer ensiling period for 100 days enhanced the fermentation characteristics of silage compared to ensiling for 50 days.

Immunohistochemical Assessment of E-cadherin and β-catenin in the Histological Differentiations of Oral Squamous Cell Carcinoma

  • Zaid, Khaled Waleed
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8847-8853
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    • 2014
  • The aim of this study was to establish the expression and localization of E-cadherin and ${\beta}$-catenin in oral squamous cell carcinomas (OSCC) so that we could correlate the findings with prognostic-relevant histopathological variables. E-cadherin and ${\beta}$-catenin expression in normal oral epithelia and in oral squamous cell carcinomas was examined immunohistochemically, and associations with histopathological differentiation and prognosis were then analyzed in 33 patients who had been operated on for OSCC. E-cadherin expression was found in (82%) of the squamous cells of well differentiated OSCC, (61%) of moderately differentiated and (39%) of poorly differentiated. E-cadherin expression was significantly associated with histological grade (p=0.000). No nuclear staining was detected. In (19.5%) of the cells E-cadherin localized in the cytoplasm, with no correlation to the histological grade (p=0.106). ${\beta}$-Catenin expression was found in 87% of the squamous cells of well differentiated OSCC, 67% of moderately differentiated and 43% of poorly differentiated, the expression was significantly associated with histological grade (p=0.000). the nuclear ${\beta}$-Catenin expression appeared in 3.3% of the cells and it was correlated to the histological grade (p=0.000). In (23.5%) of the cells ${\beta}$-Catenin localized in the cytoplasm, with correlation to the histological grade (p=0.002). According to this study the expression of ${\beta}$-catenin and E-cadherin were independent prognostic factors for histological grade. E-cadherin was closely linked to ${\beta}$-catenin expression in OSCC (p=0.000) and to tumor differentiation. That reflects a structural association and the role of both in tumor progression.

The relationship between job satisfaction and intention to quit of employees in F&B department of hotels by job type (직무 유형에 따른 특급호텔 조리ㆍ식음료 종사원의 직무만족도가 이직의사 견해에 미치는 영향 분석)

  • 양일선;명미선;이소정;이해영
    • Korean journal of food and cookery science
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    • v.20 no.2
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    • pp.151-157
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    • 2004
  • This study was designed firstly to measure the job satisfaction levels of F&B (Food & Beverage) personnel in hotels by using JDI (Job Descriptive Index) and secondly to identify the effects of job satisfaction on intention to quit by job type. Of 498 respondents, there were 245 employees in the back of the house (eBOH) and 253 in the front of the house (eFOH). From an analysis on job satisfaction, total JDI, score was 136.57${\pm}$11.48 out of a maximum 210 and the payroll (14.79 out of 21, 70.4%) facet of JDI was the highest job satisfaction followed by co-workers (36.07 out of 54, 66.8%), supervision (35.37 out of 54, 65.5%), promotion (17.45 out of 27, 64.6%) and work itself (32.86 out of 54, 60.8%). In comparison between the job satisfaction of eBOH and eFOH, eFOH perceived significantly higher than eBOH for job itself (p<.001) and total job satisfaction (p<.01). No facets of JDI were significant by job level, but work itself (p<.05), payroll (p<.05), supervision (p<.01) and promotion (p<.05) were significant by work section in eFOH,. In addition, no facets except work itself were significant by either job level or work section in eFOH. Finally, among job satisfaction facets, payroll had a negative effect on intention to quit whereas supervision had a positive effect on it in eBOH. On the other, work itself affected intention to quit positively but promotion affected it negatively. The findings confirmed the necessity for internal marketing focusing on the lower level of job satisfaction facets and indicated that such a focus would improve organizational effectiveness.

Characterization of the Lytic Bacteriophage phiEaP-8 Effective against Both Erwinia amylovora and Erwinia pyrifoliae Causing Severe Diseases in Apple and Pear

  • Park, Jungkum;Lee, Gyu Min;Kim, Donghyuk;Park, Duck Hwan;Oh, Chang-Sik
    • The Plant Pathology Journal
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    • v.34 no.5
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    • pp.445-450
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    • 2018
  • Bacteriophages, bacteria-infecting viruses, have been recently reconsidered as a biological control tool for preventing bacterial pathogens. Erwinia amylovora and E. pyrifoliae cause fire blight and black shoot blight disease in apple and pear, respectively. In this study, the bacteriophage phiEaP-8 was isolated from apple orchard soil and could efficiently and specifically kill both E. amylovora and E. pyrifoliae. This bacteriophage belongs to the Podoviridae family. Whole genome analysis revealed that phiEaP-8 carries a 75,929 bp genomic DNA with 78 coding sequences and 5 tRNA genes. Genome comparison showed that phiEaP-8 has only 85% identity to known bacteriophages at the DNA level. PhiEaP-8 retained lytic activity up to $50^{\circ}C$, within a pH range from 5 to 10, and under 365 nm UV light. Based on these characteristics, the bacteriophage phiEaP-8 is novel and carries potential to control both E. amylovora and E. pyrifoliae in apple and pear.

Modigication of host cells and Expression of Recombinant E. coli trp plasmids for the increased Production of Tryptophan in Klebsiella pneumoniae (Klebsiella pneumoniae에서 트립토판 생산증대를 위한 숙주개발 및 재조합 trp plasmid의 발현)

  • 지연태;홍광원;박장현;이세영
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.46-51
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    • 1987
  • In order to increase the production of tryptophan by maximizing expression of recombinant trp plasmid, Klebsiella pneumoniae KC 105(pheA tyrA trpE trpR tyrR) was genetically modified. KC 107, inosine monophospate(IMP) auxotroph from KC 105 and KC 108, histidine(His) auxotroph from KC 107 were also derived respectively to increase phosphoribosylpyrophosphate(PRPP) production which is required for tryptophan biosynthesis. From KC 107 phosphoribosylpyrophosphate consumption which is required for tryptophan biosynthesis. From KC 107 and KC 108, KC 109 and KC 110, both arginine auxotrophs were derived respectively. To investigate the expression of recombinant trp plasmid in the selected K. pneumoniae mutants, the auxotrophic mutants were transformed with recombinant trp plasmids pSC 101-$trpE^{FBR}$, pSC 101-trpL(.DELTA.att) $trpE^{FBR}$ (pSC 101-trp-AF). Amount of tryptophan produced and activities of tryptophan synthase of $trpR^{-}$ mutant (KC 100) and $tyrR^{-}$ mutnat(KC 105) containing recombinant plasmid pSC 101-trp operon were increased by 30-40% as compared with KC 99(pheA tyrA trpE) containing recombinant plasmid pSC 101-trp operon. Activities of tryptophan synthase and production of tryptophan of KC 108 ($His^{-}$) and KC 109($Arg^{-}$) containing recombinant plasmid pSC 101-trp operon were increase by two-fold as compared with KC 107 containing pSC 101-trp operon.

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Tumor Suppressor Protein p53 Promotes 2-Methoxyestradiol-Induced Activation of Bak and Bax, Leading to Mitochondria-Dependent Apoptosis in Human Colon Cancer HCT116 Cells

  • Lee, Ji Young;Jee, Su Bean;Park, Won Young;Choi, Yu Jin;Kim, Bokyung;Kim, Yoon Hee;Jun, Do Youn;Kim, Young Ho
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1654-1663
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    • 2014
  • To examine the effect of tumor suppressor protein p53 on the antitumor activity of 2-methoxyestradiol (2-MeO-$E_2$), 2-MeO-$E_2$-induced cell cycle changes and apoptotic events were compared between the human colon carcinoma cell lines HCT116 ($p53^{+/+}$) and HCT116 ($p53^{-/-}$). When both cell types were exposed to 2-MeO-$E_2$, a reduction in the cell viability and an enhancement in the proportions of $G_2/M$ cells and apoptotic sub-$G_1$ cells commonly occurred dose-dependently. These 2-MeO-$E_2$-induced cellular changes, except for $G_2/M$ arrest, appeared to be more apparent in the presence of p53. Immunofluorescence microscopic analysis using anti-${\alpha}$-tubulin and anti-lamin B2 antibodies revealed that after 2-MeO-$E_2$ treatment, impaired mitotic spindle network and prometaphase arrest occurred similarly in both cell types. Following 2-MeO-$E_2$ treatment, only HCT116 ($p53^{+/+}$) cells exhibited an enhancement in the levels of p53, p-p53 (Ser-15), $p21^{WAF1/CIP1}$, and Bax; however, the Bak level remained relatively constant in both cell types, and the Bcl-2 level decreased only in HCT116 ($p53^{+/+}$) cells. Additionally, mitochondrial apoptotic events, including the activation of Bak and Bax, loss of ${\Delta}{\psi}m$, activation of caspase-9 and -3, and cleavage of lamin A/C, were more dominantly induced in the presence of p53. The Bak-specific and Bax-specific siRNA approaches confirmed the necessity of both Bak and Bax activations for the 2-MeO-$E_2$-induced apoptosis in HCT116 cells. These results show that among 2-MeO-$E_2$-induced apoptotic events, including prometaphase arrest, up-regulation of Bax level, down-regulation of Bcl-2 level, activation of both Bak and Bax, and mitochondria-dependent caspase activation, the modulation of Bax and Bcl-2 levels is the target of the pro-apoptotic action of p53.

Kinetics and Mechanism of the Selective Oxidation of Ethylene for Ethylene Oxide over Monolithic Silver Catalyst (모놀리스형 은촉매상에서 에틸렌선택산화반응의 속도론적 고찰)

  • Park, Rho-Bum;Kim, Sang-Chai;Sunwoo, Chang-Sin;Yu, Eui-Yeon
    • Applied Chemistry for Engineering
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    • v.2 no.2
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    • pp.165-174
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    • 1991
  • The kinetics and the mechanism for the selective oxidation of ethylene on the supported monolithic silver catalyst were experimentally investigated in a fixed bed tubular reactor. The formation rates of ethylene oxide and carbon dioxide were measured at the atmospheric pressure with various combinations of partial pressures of ethylene and oxygen at temperature range of $225-300^{\circ}C$, conversion with 1.2-7.5 %, and then the mechanism of the selective oxidation of ethylene was verified. Their formation rates fitted with the Langmuir-Hinshelwood mechnism. The ethylene oxide and carbon dioxide are produced by reation of adsorbed ethylene with monoatomic oxygen adsorbed on the active sites of Ag-surface, and their formation rate equation are expressed as : $R_{EO}={\frac{k_1K_0{^{1/2}}K_EK_SP_{02}{^{3/2}}P_E}{(1+{\sqrt{K_0P_{02}}}+K_EP_E+K_PP_P)^2(1+{\sqrt{K_SP_{02}})^2}}$ $R_C={\frac{k_2K_0{^3}K_EK_S{^{7/2}}P_{02}{^{13/2}}P_E}{(1+{\sqrt{K_0P_{02}}}+K_EP_E+K_PP_P)^7(1+{\sqrt{K_SP_{02}})^7}}$ The activation energies of ethylene oxide and dioxide and carbon dioxide formations can be estimated to be 12.25 and 17.85 Kcal/mol, respectively.

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