• Title/Summary/Keyword: Crude extracts

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Sageretia thea fruit extracts rich in methyl linoleate and methyl linolenate downregulate melanogenesis via the Akt/GSK3β signaling pathway

  • Ko, Gyeong-A;Shrestha, Sabina;Cho, Somi Kim
    • Nutrition Research and Practice
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    • v.12 no.1
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    • pp.3-12
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    • 2018
  • BACKGROUND/OBJECTIVES: Sageretia thea is traditionally used as a medicinal herb to treat various diseases, including skin disorders, in China and Korea. This study evaluated the inhibitory effect of Sageretia thea fruit on melanogenesis and its underlying mechanisms in B16F10 mouse melanoma cells. The active chemical compounds in anti-melanogenesis were determined in Sageretia thea. MATERIALS/METHODS: Solvent fractions from the crude extract were investigated for anti-melanogenic activities. These activities and the mechanism of anti-melanogenesis in B16F10 cells were examined by determining melanin content and tyrosinase activity, and by performing western blotting. RESULTS: The n-hexane fraction of Sageretia thea fruit (HFSF) exhibited significant anti-melanogenic activity among the various solvent fractions without reducing viability of B16F10 cells. The HFSF suppressed the expression of tyrosinase and tyrosinase-related protein 1 (TRP1). The reduction of microphthalmia-associated transcription factor (MITF) expression by the HFSF was mediated by the Akt/glycogen synthase kinase 3 beta ($GSK3{\beta}$) signaling pathway, which promotes the reduction of ${\beta}-catenin$. Treatment with the $GSK3{\beta}$ inhibitor 6-bromoindirubin-3'-oxime (BIO) restored HFSF-induced inhibition of MITF expression. The HFSF bioactive constituents responsible for anti-melanogenic activity were identified by bioassay-guided fractionation and gas chromatography-mass spectrometry analysis as methyl linoleate and methyl linolenate. CONCLUSIONS: These results indicate that HFSF and its constituents, methyl linoleate and methyl linolenate, could be used as whitening agents in cosmetics and have potential for treating hyperpigmentation disorders in the clinic.

Analysis of Nutritional Characteristics and Physiological Functionality of Hypsizygus marmoreus (Brown cultivar) (갈색 느티만가닥버섯의 영양특성 및 생리기능성 분석)

  • Bolormaa, Zanabaatar;Kang, Min-Gu;Seo, Geon-Sik;Lee, Young-Wook;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.40 no.2
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    • pp.104-108
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    • 2012
  • In order to apply into functional food or medicinal industry, nutritional characteristics and physiological functionality of Hypsizygus marmoreus (brown cultivar) were investigated. Fruiting body of H. marmoreus contained 27.3% of crude protein, 55.8% of total sugar and 11,109.3 mg/100 g dry weight of malic acid. Furthermore, 66.7% of antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity and 37.3% of antigout xanthin oxidase inhibitory activity showed in the water extract from H. marmoreus. The economically ACE inhibitory activity (81.4%) was obtained when the fruiting body of Hysizygus marmoreus was extracted with distilled water of $50^{\circ}C$ for 12 h, even though maximal ACE inhibitory activity (84.4%) was showed the extracts from $70^{\circ}C$ for 12 h.

Construction of Genetically Engineered Microorganisms for Overexpression of xylE Gene Encoding Catechol 2,3-dioxygenase and the Functional Stability of the Recombinant Plasmid pSW3a Containing xylE in Aquatic Environment

  • Han, Hyo-Yung;Kim, Chi-Kyung;Park, Yong-Keun;Ka, Jong-Ok;Lee, Byeong-Jae;Min, Kyung-Hee
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.341-348
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    • 1996
  • The regulation of xylE gene expression was examined by using vector promoter and construction of genetically engineered microorganisms (GEMs) for application in microcosm. When the xylE gene wsa subcloned into pBluscript SK(+) under the control of lac promoter (pTY1) in E. coli, and the expression was induced by IPTG, the enzyme activity of catechol 2, 3-dioxygenase was increased 4.7 times more than that of the crude extracts from transformants harboring pTY1. We suggest that the xylE gene has its own promoter at the upstream portion, because it was able to be expressed even in the absence of IPTG. A recombinant plasmid, pSW3a harboring the xylE gene under the T7 promotor, showed the activity of 14.5 units/mg protein, higher than that of parental strain, E. coli PYT1. The xylE gene in recombinant plasmid pSW3a was used as reporter gene for the application in microcosm ecosystem, since it was used for detection of xylE-positive clones by catechol spray on the agar plates. The pSW3a in E. coli was introduced into Pseudomonas patida to construct GEM strain, and examined for the exxpression and functional stability in microcosms.

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Extraction of Glycyrrhizic Acid from Licorice using Supercritical Carbon Dioxide/Aqueous Ethanol (초임계 이산화탄소/에탄올을 이용한 감초의 Glycyrrhizic acid 추출)

  • 김현석;김병용;이상윤;김우식;이은규;유종훈;임교빈
    • KSBB Journal
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    • v.18 no.5
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    • pp.347-351
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    • 2003
  • The extraction of glycyrrhizic acid from licorice using supercritical carbon dioxide (SCCO$_2$) was investigated with respect to the effects of extraction parameters such as the kind and amount of modifier, temperature, pressure, and extraction time. The conventional organic solvent extraction was also conducted for a quantitative comparison. The content of glycyrrhizic acid in crude extracts was analyzed by HPLC and the yield of glycyrrhizic acid was computed as a weight percent recovery. The optimal pressure and temperature for SCCO$_2$ extraction were found to be 40 MPa and 80$^{\circ}C$, respectively, when SCCO$_2$ was modified with 70% aqueous ethanol. Under the same pressure and temperature, the highest recovery was attained to be 104.57% in the first 60 min when the concentration of 60%, aqueous ethanol in SCCO$_2$ was 15%.

Studies on the Compositions of Amino acid and Fatty acid of Pine caterpillar(Dendrolimus spectabilis) (송충이 단백질의 아미노산과 지방산 조성에 대한 연구)

  • 차월석;신주연김연순
    • KSBB Journal
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    • v.7 no.3
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    • pp.186-190
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    • 1992
  • The compositions of amino acid in the protein and total fatty acid of Dendrolimus spectabilis were analyzed quantitatively by HPLC and GC, respectively. The contents of crude oil and protein from the extracts were 21.00% and 58.47%, respectively. The amount of free amino acids in the protein was 3.65g/100g, and 1.31g/100g of essential amino acids was contained in the free amino acids. The amount of total amino acids in the protein was 41.20g/100g, and 14.75g/100g of essential amino acids were contained in the total amino acid. The compositions of fatty acid in the oil were $C_{18}$=26.81%, $C_{16}$=19.09%, $C_{18:1}$=18.74%, $C_{18:3}$=15.33%, $C_{16:1}$=7.29%, $C_{20}$=5.21% in order, respectively. 45.88% of unsaturated fatty acids were contained in the oil.

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Genetically Engineered Biosynthesis of Macrolide Derivatives Including 4-Amino-4,6-Dideoxy-L-Glucose from Streptomyces venezuelae YJ003-OTBP3

  • Pageni, Binod Babu;Oh, Tae-Jin;Liou, Kwang-Kyoung;Yoon, Yeo-Joon;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.88-94
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    • 2008
  • Two sugar biosynthetic cassette plasm ids were used to direct the biosynthesis of a deoxyaminosugar. The pOTBP1 plasmid containing TDP-glucose synthase (desIII), TDP-glucose-4,6-dehydratase (desIV), and glycosyltransferase (desVII/desVIII) was constructed and transformed into S. venezuelae YJ003, a strain in which the entire gene cluster of desosamine biosynthesis is deleted. The expression plasmid pOTBP3 containing 4-aminotransferase (gerB) and 3,5-epimerase (orf9) was transformed again into S. venezuelae YJ003-OTBP1 to obtain S. venezuelae YJ003-OTBP3 for the production of 4-amino-4,6-dideoxy-L-glucose derivatives. The crude extracts obtained from S. venezuelae ATCC 15439, S. venezuelae YJ003, and S. venezuelae YJ003-OTBP3 were further analyzed by TLC, bioassay, HPLC, ESI/MS, LC/MS, and MS/MS. The results of our study clearly shows that S. venezuelae YJ003-OTBP3 constructs other new hybrid macrolide derivatives including 4-amino-4,6-dideoxy-L-glycosylated YC-17 (3, [M+ $Na^+$] m/z=464.5), methymycin (4, m/z=480.5), novamethymycin (6, m/z=496.5), and pikromycin (5, m/z=536.5) from a 12-membered ring aglycon (10-deoxymethynolide, 1) and a 14-membered ring aglycon (narbonolide, 2). These results suggest a successful engineering of a deoxysugar pathway to generate novel hybrid macrolide derivatives, including deoxyaminosugar.

Effect of Transgenic Rhizobacteria Overexpressing Citrobacter braakii appA on Phytate-P Availability to Mung Bean Plants

  • Patel, Kuldeep J.;Vig, Saurabh;Nareshkumar, G.;Archana, G.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1491-1499
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    • 2010
  • Rhizosphere microorganisms possessing phytase activity are considered important for rendering phytate-phosphorus (P) available to plants. In the present study, the Citrobacter braakii phytase gene (appA) was overexpressed in rhizobacteria possessing plant growth promoting (PGP) traits, for increasing their potential as bioinoculants. AppA was cloned under the lac promoter in the broadhost-range expression vector pBBR1MCS-2. Transformation of the recombinant construct pCBappA resulted in high constitutive phytase activity in all of the eight rhizobacterial strains belonging to genera Pantoea, Citrobacter, Enterobacter, Pseudomonas (two strains), Rhizobium (two strains), and Ensifer that were studied. Transgenic rhizobacterial strains were found to display varying levels of phytase activity, ranging from 10-folds to 538-folds higher than the corresponding control strains. The transgenic derivative of Pseudomonas fluorescens CHA0, a well-characterized plant growth promoting rhizobacterium, showed the highest expression of phytase (~8 U/mg) activity in crude extracts. Although all transformants showed high phytase activity, rhizobacteria having the ability to secrete organic acid showed significantly higher release of P from Ca-phytate in buffered minimal media. AppA overexpressing rhizobacteria showed increased P content, and dry weight (shoot) or shoot/ root ratio of mung bean (Vigna radiata) plants, to different extents, when grown in semisolid agar (SSA) medium containing Na-phytate or Ca-phytate as the P sources. This is the first report of the overexpression of phytase in rhizobacterial strains and its exploitation for plant growth enhancement.

Development of Indole-3-Acetic Acid-Producing Escherichia coli by Functional Expression of IpdC, AspC, and Iad1

  • Romasi, Elisa Friska;Lee, Jinho
    • Journal of Microbiology and Biotechnology
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    • v.23 no.12
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    • pp.1726-1736
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    • 2013
  • Biosynthesis of indole-3-acetic acid (IAA) via the indole-3-pyruvic acid pathway involves three kinds of enzymes; aminotransferase encoded by aspC, indole-3-pyruvic acid decarboxylase encoded by ipdC, and indole-3-acetic acid dehydrogenase encoded by iad1. The ipdC from Enterobacter cloacae ATCC 13047, aspC from Escherichia coli, and iad1 from Ustilago maydis were cloned and expressed under the control of the tac and sod promoters in E. coli. According to SDS-PAGE and enzyme activity, IpdC and Iad1 showed good expression under the control of $P_{tac}$, whereas AspC was efficiently expressed by $P_{sod}$ originating from Corynebacterium glutamicum. The activities of IpdC, AspC, and Iad1 from the crude extracts of recombinant E. coli Top 10 were 215.6, 5.7, and 272.1 nmol/min/mg-protein, respectively. The recombinant E. coli $DH5{\alpha}$ expressing IpdC, AspC, and Iad1 produced about 1.1 g/l of IAA and 0.13 g/l of tryptophol (TOL) after 48 h of cultivation in LB medium with 2 g/l tryptophan. To improve IAA production, a tnaA gene mediating indole formation from tryptophan was deleted. As a result, E. coli IAA68 with expression of the three genes produced 1.8 g/l of IAA, which is a 1.6-fold increase compared with wild-type $DH5{\alpha}$ harboring the same plasmids. Moreover, the complete conversion of tryptophan to IAA was achieved by E. coli IAA68. Finally, E. coli IAA68 produced 3.0 g/l of IAA after 24 h cultivation in LB medium supplemented with 4 g/l of tryptophan.

Properties of Crude $\beta$-amylase from Korean ginseng, Panax ginseng C. A. Meyer (고려인삼(Panax Ginseng C. A. Meyer) 중의 조(組) $\beta$-amylase의 분리와 그 성질)

  • Kim, Byung-Mook
    • Korean Journal of Food Science and Technology
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    • v.17 no.4
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    • pp.240-244
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    • 1985
  • Curde $\beta$-amylase was prepared by frationation of the water extracts from Korean ginseng, Panax Ginseng C.A. Meyer, with 0.2-0.6 saturation of ammonium sulfate. The enzyme showed the typical properties of $\beta$-amylase, producing only maltase from starch. The enzyme preparation also showed no maltase activity. The enzyme was stable at the pH 5-9 and at the temperature below $40^{\circ}C$. The enzyme showed the optimum pH at 5.0 and the optimum temperature at $35^{\circ}C$. Its activities had proportional relations with substrate concentration below 12 mg%, showing Km V slues of 4.76 mg%. The enzyme was inhibited by $Ag^{+}$, $Hg^{++}$, $Cd^{++}$, $Cu^{++}$,$ Al^{3+}$, and $Fe^{3+}$.

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The Enhanced Effect of Oplopanax elatus Nakai on the Immune System and Antitumor Activity (땃두릅(Oplopanax elatus Nakai) 추출물의 면역자극 활성 및 항암 증진 효과)

  • Hur, Jin Woo;Cho, Eun Hee;Lee, Bo Kyung;Lee, Uiyoung;Yoon, Taek Joon
    • The Korean Journal of Food And Nutrition
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    • v.26 no.3
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    • pp.375-382
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    • 2013
  • The present study is designed to explore an anti-tumor activity on crude extracts of Oplopanax elatus. Water extractions of Oplopanax elatus were performed at $100^{\circ}C$(OeE-100). OeE-100 doses up to $62.5{\mu}g/m{\ell}$ had no cytotoxicity on the tumor cell lines in vitro. In experimental lung metastasis of colon26-M3.1 carcinoma or B16-BL6 melanoma, the prophylactic intravenous ($4{\sim}100{\mu}g/mouse$) or oral (2 mg/mouse) administration of OeE-100 significantly inhibited tumor metastasis as compared with tumor controls. Peritoneal macrophages stimulated with OeE-100 produced various cytokines such as TNF-${\alpha}$, IL-6 and IL-12. In an analysis of NK-cell activities, i.v. administration of OeE-100 ($10{\sim}100{\mu}g/mouse$) significantly augmented the cytotoxicity to YAC-1 tumor cells. Vaccination of mice with boiling-treated tumor cells (BT-vaccine) in combination with OeE-100 ($100{\mu}g/mouse$) showed higher inhibitions in tumor metastasis when compared with the mice of BT-vaccine treatment. In addition, the splenocytes from OeE-100 admixed BT-vaccine immunized mice secreted a higher concentration of Th1 type cytokine such as IFN-${\gamma}$. These results suggested that the OeE-100 stimulated immune system and was a good candidate adjuvant of anti-tumor immune responses.