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Production of the Rare Ginsenoside Rh2-MIX (20(S)-Rh2, 20(R)-Rh2, Rk2, and Rh3) by Enzymatic Conversion Combined with Acid Treatment and Evaluation of Its Anti-Cancer Activity

  • Song, Bong-Kyu;Kim, Kyeng Min;Choi, Kang-Duk;Im, Wan-Taek
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1233-1241
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    • 2017
  • The ginsenoside Rh2 has strong anti-cancer, anti-inflammatory, and anti-diabetic effects. However, the application of ginsenoside Rh2 is restricted because of the small amounts found in Korean white and red ginsengs. To enhance the production of ginsenoside Rh2-MIX (comprising 20(S)-Rh2, 20(R)-Rh2, Rk2, and Rh3 as a 10-g unit) with high specificity, yield, and purity, a new combination of enzymatic conversion using the commercial enzyme Viscozyme L followed by acid treatment was developed. Viscozyme L treatment at pH 5.0 and $50^{\circ}C$ was used initially to transform the major ginsenosides Rb1, Rb2, Rc, and Rd into ginsenoside F2, followed by acid-heat treatment using citric acid 2% (w/v) at pH 2.0 and $121^{\circ}C$ for 15 min. Scale-up production in a 10-L jar fermenter, using 60 g of the protopanaxadiol-type ginsenoside mixture from ginseng roots, produced 24 g of ginsenoside Rh2-MIX. Using 2 g of Rh2-MIX, 131 mg of 20(S)-Rh2, 58 mg of 20(R)-Rh2, 47 mg of Rk2, and 26 mg of Rh3 were obtained at over 98% chromatographic purity. Then, the anti-cancer effect of the four purified ginsenosides was investigated on B16F10, MDA-MB-231, and HuH-7 cell lines. As a result, these four rare ginsenosides markedly inhibited the growth of the cancer cell lines. These results suggested that rare ginsenoside Rh2-MIX could be exploited to prepare an anti-cancer supplement in the functional food and pharmaceutical industries.

Studies on the Amylase Production by Bacteria (세균(細菌)에 의(依)한 Amylase생산(生産)에 관한 연구(硏究))

  • Park, Yoon-Joong
    • Applied Biological Chemistry
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    • v.13 no.2
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    • pp.153-170
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    • 1970
  • 1. Isolation and identification of amylase-producing bacteria. The powerful strain A-12 and S-8 were respectively isolated from air and soil after screening a large number of amylase-producing bacteria. Their bacterial characteristics have been investigated and it has been found that all characteristics of strain A-12 and S-8 are similar to Bac. subtilis of Bergey's manual except for the acid formation from a few carbohydrates and the citrate utilization, i.e., the strain A-12 shows negative in the citrate utilization, and the acid formation from arabinose and xylose, S-8 shows negative in the acid formation from xylose. 2. Amylase production by Liquid cultures with solid materials. Several conditions for amylase production by strain A-12 in stationary cultures have been studied. The results obtained are as follows. (1) The optimum conditions are:temperature $35^{\circ}C$, initial pH 6.5 to 7.0 and incubation time 3 to 4 days. (2) The amylase production is not affected by the preservation period of the stock cultures. (3) Among the various solid material, the defatted soy bean is found to be the best for t1e amylase production. However, the alkali treatment of the defatted soy bean gives no effect contrary to the cage of defatted rape seed. The addition of soluble starch to the alkali extract of defatted soy bean shows the increased amylase production. (4) Up to 1% addition of ethanol to carbon dificient media gives the improved amylase production, whereas the above effect is not found in the case of carbon rich media. (5) The amylase production can be increased 2.5 times when 10% of defatted soy bean is admixed to cheaply available wheat bran. (6) The excellent effect is found for amylase production when 20% of wheat bran is admixed to defatted dry milk which is a poor medium. The activity is found to be $D^{40^{\circ}}_{30'}$ 7,000(L.S.V. 1,800) in 10% medium. (7) No significant effect is observed due to the addition of various inorganic salts. 3. Amylase production by solid cultures. Several conditions for amylase production by strain A-12 in wheat bran cultures have been studied and the results obtained are as follows. (1) The optimum conditions: are temperature $33^{\circ}C$, incubation lime 2 days, water content added 150 to 175% and the thickness of the medium 1.5cm, The activity is found to be $D^{40^{\circ}}_{30'}$ 36,000(L.S.V. 15,000) (2) No significant effect is found in the case of the additions of various organic and inorganic substances.

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Physiological Effect of Yogurt with Powder Two Stage Fermented Dioscorea batatas Dence by Monascus sp. and Lactobacillus sp. (Monascus sp.와 Lactobacillus sp.를 이용한 2단 발효마 분말 첨가 요구르트의 생리활성 효과)

  • Jeon, Chun-Pyo;Lee, Jung-Bok;Choi, Chung-Sig;Kwon, Gi-Seok
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.151-157
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    • 2011
  • This study was to examine the quality characteristics and biological activities of the yogurt containing dual fermentation MFCY (Monascus-Fermented Chinese Yam) powder. Six different contents (0, 0.1, 0.3, 0.5, 1.0, and 2.0%, w/v) of MFCY powder were added to raw milk and 5% skim milk followed by fermentation with lactic acid bacteria at $37^{\circ}C$ for 12 h, and then the biological activities of the samples were investigated. Biological activities of the prepared yogurt were evaluated for acid production (pH, titratable acidity), number of viable cells, total polyphenol contents, DPPH radical scavenging activity, reducing power, angiotensin converting enzyme inhibitory effects and GABA contents. In this study, the results show that after fermentation in raw milk and 5% skim milk added with MFCY powder. The yogurt can produced pH, titratable acidity, number of viable cells, total polyphenol contents, DPPH radical scavenging activity and reducing power at 4.12-4.25 (pH), 0.94-0.97% (TA), $7.50{\times}10^9-1.14{\times}10^{10}$ CFU/ml, 161.4-329.9 (TP, mg/L), 54-94% (DPPH), and 1.13-1.53 (R.P), respectively. In addition, the showed ACE inhibitory activity and GABA contents was 67.1-87.7% and 304.6-685.4 (mg/L).

Solution Processed Porous Fe2O3 Thin Films for Solar-Driven Water Splitting

  • Suryawanshi, Mahesh P.;Kim, Seonghyeop;Ghorpade, Uma V.;Suryawanshi, Umesh P.;Jang, Jun Sung;Gang, Myeng Gil;Kim, Jin Hyeok;Moon, Jong Ha
    • Korean Journal of Materials Research
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    • v.27 no.11
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    • pp.631-635
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    • 2017
  • We report facile solution processing of mesoporous hematite (${\alpha}-Fe_2O_3$) thin films for high efficiency solar-driven water splitting. $Fe_2O_3$ thin films were prepared on fluorine doped tin oxide(FTO) conducting substrates by spin coating of a precursor solution followed by annealing at $550^{\circ}C$ for 30 min. in air ambient. Specifically, the precursor solution was prepared by dissolving non-toxic $FeCl_3$ as an Fe source in highly versatile dimethyl sulfoxide(DMSO) as a solvent. The as-deposited and annealed thin films were characterized for their morphological, structural and optical properties using field-emission scanning electron microscopy(FE-SEM), X-ray diffraction(XRD), X-ray photoelectron spectroscopy(XPS) and UV-Vis absorption spectroscopy. The photoelectrochemical performance of the precursor (${\alpha}-FeOOH$) and annealed (${\alpha}-Fe_2O_3$) films were characterized and it was found that the ${\alpha}-Fe_2O_3$ film exhibited an increased photocurrent density of ${\sim}0.78mA/cm^2$ at 1.23 V vs. RHE, which is about 3.4 times higher than that of the ${\alpha}-FeOOH$ films ($0.23mA/cm^2$ at 1.23 V vs. RHE). The improved performance can be attributed to the improved crystallinity and porosity of ${\alpha}-Fe_2O_3$ thin films after annealing treatment at higher temperatures. Detailed electrical characterization was further carried out to elucidate the enhanced PEC performance of ${\alpha}-Fe_2O_3$ thin films.

Purification and Enzymatic Characteristics of the Bacillus pasteurii Urease Expressed in Escherichia coli (Escherichia coli에서 발현된 Recombinant Bacillus pasteurii Urease의 정제 및 효소학적 특성)

  • 이은탁;김상달
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.519-526
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    • 1992
  • The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

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Effects of Various Proteins on the Autoxidation of L-Ascorbic Acid (비타민 C 산화반응에 대한 단백질의 공존효과)

  • Kim Mi-Ok;Jang Sang-Moon
    • The Korean Journal of Food And Nutrition
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    • v.17 no.3
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    • pp.294-301
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    • 2004
  • Effects of superoxide dismutase(SOD), catalase(CAT), and such other proteins as bovine serum albumin(BSA), ovalbumin, lysozyme, and v-globulin on the autoxidation rates of L-ascorbic acid(AsA) in the absence of heavy metal ions and in the presence of Fe(III) or Cu(II) ions in water were examined. AsA was dissolved in a ultra-refined water at a concentration of 50 ${\mu}$M and 5 ${\mu}$M Fe(III) or 0.1 ${\mu}$M Cu(II) were added, and a oxygen gas was bubbled through the solution at a flow rate of 200 ml/min at 35$^{\circ}C$. The amount of remaining AsA in the reaction mixture was determined by using a UV spectrophotometer(at 265 nm). It was found that the Cu(II) at a concentration of 0.1 ${\mu}$M had a more accelerated for the autoxidation of AsA than Fe(III) at 5 ${\mu}$M. Moreover, it was confirmed that the ratio of remaining AsA was significantly larger in the presence of SOD, CAT, BSA, ovalbumin, lysozyme, and v-globulin than in the absence of proteins. The stabilization of AsA by various proteins were confirmed during the autoxidation of AsA in the presence of Fe(III) or Cu(II) in water. It was suggested that the non-enzymatic effects of SOD, CAT and some other proteins might be involves in the stabilization of AsA.

The Design and Fabrication of Conversion Layer for Application of Direct-Detection Type Flat Panel Detector (직접 검출형 평판 검출기 적용을 위한 변환층 설계 및 제작)

  • Noh, Si-Cheol;Kang, Sang-Sik;Jung, Bong-Jae;Choi, Il-Hong;Cho, Chang-Hoon;Heo, Ye-Ji;Yoon, Ju-Seon;Park, Ji-Koon
    • Journal of the Korean Society of Radiology
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    • v.6 no.1
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    • pp.73-77
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    • 2012
  • Recently, Interest to the photoconductor, which is used to flat form X-ray detector such as a-Se, $HgI_2$, PbO, CdTe, $PbI_2$ etc. is increasing. In this study, the film layer by using the photoconductive material with particle sedimentation was fabricated and evaluated. The quantization efficiency of the continuous X-ray with the 70 kVp energy bandwidth was analyzed by using the Monte Carlo simulation. With the results, the thickness of film with 64 % quantization efficiency was 180 ${\mu}m$ which is similar to the efficiency of 500 ${\mu}m$ a-Se film. And $HIg_2$ film has the high quantization efficiency of 74 % on 240 ${\mu}m$ thickness. The electrical characteristics of the 239 ${\mu}m$ $Hgl_2$ films produced by particle sedimentation were shown as very low dark current(under 10 $pA/mm^2$), and high sensitivity(19.8 mC/mR-sec) with 1 $V/{\mu}m$ input voltage. The SNR, which is influence to the contrast of X-ray image, was shown highly as 3,125 in low driving voltage on 0.8 $V/{\mu}m$. With the results of this study, the development of the low-cost, high-performance image detector with film could be possible by replacing the film produced by particle sedimentation instead to a-Se detector.

In vitro Test of Mycelial Growth Inhibition of 5 Fungi Pathogenic to Strawberries by Ultraviolet-C (UV-C) Irradiation (자외선(UV-C) 조사에 의한 딸기병원균의 균사생장억제)

  • Kim, Seon Ae;Ahn, Soon-Young;Oh, Wook;Yun, Hae Keun
    • Korean Journal of Food Science and Technology
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    • v.44 no.5
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    • pp.634-637
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    • 2012
  • In strawberry production, among others, the high incidence of diseases by pathogenic fungi resulting in the reduction of fruit yield and quality requires the development of eco-friendly management systems rather than chemical sprays to control them. The diameter of colonies grown in media at $25^{\circ}C$ for 5 days was measured to evaluate the in vitro inhibition of mycelial growth of 5 pathogenic fungi by irradiation with ultraviolet (UV-C, 264 nm). The mycelial growth of 5 pathogenic fungi was inhibited in potato dextrose agar (PDA) by the irradiation of UV-C for 1 hour a day, and was dramatically inhibited by the irradiation of UV-C for 9-12 h a day. The irradiation of UV-C for 9-12 h a day inhibited completely the growth of the late blight pathogen, Phytophthora cactorum. The irradiation distance of 40 to 50 cm was effective for the inhibition of mycelial growth of fungi. The mycelial growth of fungi without pre-incubation was inhibited strongly by UV-C irradiation compared to fungi pre-incubated for 2 days without light. The mycelia growth of Colletotrichum gloeosprioides and Fusarium oxysporum was inhibited strongly by UV-C irradiation in vegetable 8 juice agar compared to PDA.

The Effect of Cellophane Film Packing on Quality of Semi-Salted and Dried Mackerel during Processing and Storage (셀로판 필름보장이 반염건고등어의 가공 및 저장중의 품질에 미치는 효과)

  • 이응호;안창범;김복규;이채한;이호연
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.2
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    • pp.139-147
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    • 1991
  • The preservative effect of cellophane film packing on the quality of semi-salted and dried mackerel was studied. The product(P) of semi-salted and dried mackerel was prepared from raw mackerel by filleting, cleaning, soaking in 15%9v/w) salt solution for 30min, draining, packing with cellophane film (PT# 300, thickness:$20{\mu}{\textrm}{m}$) and drying for 4 hrs at $40^{\circ}C$ in hot air dryer. The product (C) was also prepared without cellophane film packing after draining. The product (C) and (P) were stored at $5.0{\pm}0.5^{\circ}C$. After processing and during storage, moisture content of product (P) was higher than that of product (C), but contents of VBN(volatile basic nitrogen), amino nitrogen and TMA of product (P) on dry basis were lower than those of product (C). Viable cell count, TBA value, peroxide value and decreasing rate of polyenoic acid of product (P) were also lower than those of product (C). In sensory evaluation, the shelf life of product (C) was about 9 days and that of product (P) was about 14 days. From the results of chemical and sensory evaluation, it was concluded that cellophane film packing was a good condition for preserving the quality of semi-salted and dried mackerel.

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Alterations of Cerebral Metabolic Activation Following Electro-Acupuncture Stimulation on ST36 and LR3 Acu-Points in Rats (흰쥐의 족삼리 및 태충 전침자극에 따른 뇌대사활성의 변화)

  • Sohn, Young-Joo;Jung, Hyuk-Sang;Ku, Ja-Seung;Won, Ran;Kim, Yong-Suk;Park, Young-Bae;Sohn, Nak-Won
    • Journal of Acupuncture Research
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    • v.19 no.1
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    • pp.159-174
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    • 2002
  • Objective : The meridian theory in oriental medicine explains that each acu-point has a characteristic functional effect. It will be supposed that an acupuncture stimulation on different acu-point evokes different activation on different areas in the central nervous system(CNS) according to the meridian theory. On this supposition, our group tried the semi-quantitative [14C]2-deoxyglucose([14C]2-DG) autoradiography on the acupuncture stimulation to the hindlimb acu-points of Sprague-Dawley rats. Methods : A venous catheter for the intravenous administration of isotope was equipped in the right external jugular vein on 3 days prior to the [14C]2-DG study. On the day of the study, two acupuncture needles were inserted into the ST36(Zusanli) or LR3(Taichong) on the left hindlimb. Electro-acupuncture stimulation (2 Hz, 5 ms, 1~3 mA, 15 minutes) started just before the i.v. injection of [14C]2-DG ($25{\mu}Ci/rat$). The brain and the spinal cord were removed and processed for the [14C] 2-DG autoradiography. Results : The EA stimulation on ST36 reveals over 120% metaboilc activation in Arcuate nucleus, Anterior pretectal nucleus, Dorsal cochlear nucleus, Interposed cerebellar nucleus, and Nucleus of Darkschewitsch. The EA stimulation on LR3 reveals over 120% metaboilc activation in Lateral habenula nucleus, Medial vestibular nucleus, Ventromedial thalamic nucleus, Anteroventral thalamic nucleus, Anterior cingulate cortex, Dentate gyrus, Antero cortical amygdaloid nucleus, Anterior pretectal nucleus, and Dorsal tegmental nucleus compared with the non EA stimulation control group. Conclusion : These results demonstrate that the different acu-points evoke the different activations in brain areas. And with this functional brain mapping study, a new scientific elucidation for the basis of the acupuncture-meridian theory in oriental medicine through differences of activated area in CNS according to the each acupuncture point.

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