• Title/Summary/Keyword: $\beta$-D-galactosidase

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The Inhibitory Effect of Grapefruit Seed Extracts on the Physiological Function of Enterobacter pyrinus (Grapefruit 종자추출물이 Enterobacter pyrinus의 생리기능에 미치는 영향)

  • Lee, Tae-Ho;Jeong, Sook-Jung;Lee, Sang-Yeol;Kim, Jae-Won;Cho, Sung-Hwan
    • Korean Journal of Food Science and Technology
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    • v.27 no.6
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    • pp.985-990
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    • 1995
  • Grapefruit seed extracts(GFSE) have some unknown compounds which exhibit the antibiotic activities aganist microorganisms including bacteria and fungi. We have examined the effects of GFSE on the growth of Enterobacter pyrinus which was isolated from necrotic lesions of pear trees. During the cultivation, the growth of the bacteria was strongly inhibited at the low concentration(0.01%, w/w) of GFSE. Hydrophobic fraction extracted from GFSE by mixed solvents (chloroform : methanol : water, 1 : 2 : 0.8, v/v/v) had components which inhibited the growth of bacteria. There was, however, no inhibitory effect of GFSE on the activities of several enzymes including hexokinase, glucose 6-phosphate dehydrogenase, malate dehydrogenase and succinate dehydrogenase. $O-nitrophenyl-{\beta}-D-galactopyranoside(ONPG)$, the artificial substrate of ${\beta}-galactosidase$ was hydrolyzed in the presence of GFSE, indicating that the membrane was pertubated by the GFSE. From the results it was suggested that the antibiotic activity of GFSE is due to the change of membrane permeability of cell. GFSE was fractionated by high performance liquid chromatography equipped with $C_{18}$ reverse phase column. Among active fractions, three peaks were identified as 1-chloro-2-methyl-benzene (o-toluene), N,N-dimethyl-benzenemethaneamine, 1-[2-(2-ethylethoxy)ethoxy]-4- (1,1,3,3-tetramethyl)-bezene, respectively, while the other three remained unidentified.

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Development of Cosmetic Ingredient by Fermented Paprika Juice (파프리카 발효즙의 화장품 소재개발 연구)

  • Bae, Soo Jung;Song, Min Hyeon;Oh, Jung Young;Bae, Jun Tae;Kim, Jin Hwa;Lee, Geun Soo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.2
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    • pp.117-124
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    • 2018
  • In this study, cosmetic materials were developed using a new method of making juice through the fermentation of raw natural materials with microorganisms in order to supplement the advantages and disadvantages of an organic solvent extraction method and a microbial fermentation method. The natural products were selected from two colors (red, green) of paprika known to be rich in various colors and vitamins. The microorganisms used for fermentation were fermented by inoculating paprika with lactic acid bacteria (Lactobacillus plantarum) having sugar-hydrolyzed ability. First, we investigated the changes of physiologically active substances of two kinds of paprika juice and two kinds of fermented paprika juice. Total phenols content and total flavonoids content were higher in the fermented paprika juice than in the paprika juice, and especially in the fermented red paprika juice. Free radical scavenging effect and lipid peroxidation inhibitory effect were also showed an excellent antioxidative effect on paprika fermented juice, among which the effect of red paprika fermentation juice was the highest. The expression of MMP-1 in fermented red paprika juice with high antioxidant activity was inhibited by concentration-dependent expression of MMP-1 mRNA and MMP-1 protein. In the glycation experiments with aging, the anti-glycation effect of fermented paprika juice was highly inhibited by the production of advanced glycation end-products (AGEs), which was closely related to the antioxidant effect. In addition, the activity of senescence-associated ${\beta}$-galactosidase (SA-${\beta}$-gal), an indicator of cell senescence, was measured using human dermal fibroblast (HDF). The results showed that the cell senescence was inhibited when the cells were treated with fermented paprika juice. In conclusion, fermented paprika juice using lactic acid bacteria showed better antioxidative and anti-aging effects than paprika juice. Among them, fermented red paprika juice has the best antioxidant and anti-aging effect and can be applied as natural new material of antioxidant and anti-aging.

Physiological Characteristics and GABA Production of Lactobacillus acidophilus RMK567 Isolated from Raw Milk (원유에서 분리한 Lactobacillus acidophilus RMK567의 GABA 생성 및 생리적 특성)

  • Lim, Sang-Dong;Kim, Kee-Sung;Do, Jeong-Ryong
    • Food Science of Animal Resources
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    • v.29 no.1
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    • pp.15-23
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    • 2009
  • In order to develop a new starter for fermented milk, 2082 bacteria were isolated from raw milk. The strain that showed excellent acid forming and ${\gamma}$-aminobutyric acid (GABA) production ($711.40{\mu}g/g$ D.W) characteristics after incubation at $37^{\circ}C$ for 18 hr was selected and identified as Lactobacillus acidophilus by the result of API carbohydrate fermentation pattern and 16S rDNA sequence. L. acidophilus RMK567 was investigated for its physiological characteristics. RMK67 strain showed good GABA production compared with commercial lactic acid bacteria. The optimum growth temperature of L. acidophilus RMK567 was $40^{\circ}C$ and cultures took 15 hr to reach pH 4.3. L. acidophilus RMK567 showed higher sensitivity to penicillin-G, novobiocin, as compared to other 14 different antibiotics. However, it showed more resistance to kanamycin, neomycin, streptomycin. It showed higher leucine arylamidase and ${\beta}$-galactosidase activities compared to 16 other enzymes. It was comparatively tolerant to bile juice and able to survive at pH 2 for 3 hr. It showed resistence to Escherichia coli, Salmonella typhimurium and Staphylococcus aureus with rates of 29.2%, 39.1% and 51.4%, respectively. Based on these and previous results, L. acidophilus RMK567 could be an excellent starter culture for fermented milk with excellent GABA contents.

Anti-aging Effect on Skin with the needles of red pine, Pinus densiflora (적송엽(赤松葉)이 피부(皮膚)의 노화(老化)에 미치는 영향(影響))

  • Park, Seong-Kyu;Lee, Jong-Chan;Ahn, Soo-Mi;Lee, Jin-Young;Kim, Youn-Joon;Hwang, Jae-Sung;Lee, Byeong-Gon;Chang, Ih-Seoup
    • The Korea Journal of Herbology
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    • v.20 no.4
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    • pp.1-10
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    • 2005
  • Objectives : We investigated the anti-aging effect on skin with the extract of the needles of red pine, Pinus densiflora. Methods : We measured various effects related to skin such as the anti-oxidant effect, the protection against ultraviolet (UV) irradiation, the inhibition of reactive oxygen species (ROS) generation, the induction of heat shock protein 70 (HSP70), the reduction of matrix metalloproteinase-2 (MMP-2) synthesis and senescent cell. Results : The results were as follows : The extract of the needles of red pine (RP) had the potent anti-oxidant effect and the ROS scavenging effect. Also RP preserved the systemic anti-oxidant enzyme system (superoxide dismutase and catalase) from UVB irradiation. RP protected the cell membrane from the damages induced by UVB irradiation. RP induced HSP70, a mediator of resistance to UVB irradiation. RP reduced the synthesis of MMP-2 induced by UVB irradiation. And RP inhibited the amount of senescent-associated (SA) ${\beta}-galactosidase$ staining, as a marker of replicative senescence. Conclusions : The results of our study indicate that the extract of the needles of red pine, Pinus densiflora, has anti-aging effects on skin.

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Antimicrobial Characteristics of Scutellariae Radix Extract (황금추출물의 항균특성)

  • 조성환;김영록
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.964-968
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    • 2001
  • It was carried out for research and development of natural antimicrobial on Scutellariae Radix extract against food-borne infection bacteria .Scutellariae Radix extract showed remarkable antimicrobial activites against Bacillus cereus, Listeria monocytogenes, Escherichia coli, and Vivro parahaemolyticus when examined by disk method, it was very stable on the wide rane of temperature and pH,.The growth rates of Bacillus cereus, Listeria monocytogenes, Escherichia coli, and Vivro parahaemolyticus were decreased at the concentration of more than 500 ppm Scutellariae Radix extract, Indicating that the minimum inhibitory concentration (MIC)of the Bacillus cereus, Listeria monocytogenes, Escherichia coli, and Vivro parahaemolyticus to Scutellariae Radix extract were around 500 ppm . The morphological changes were observed by transmission electron microscope and scanning electron microscope and the microbial cells membran was destroyed by Scutellariae Radix extract. It was identified that the membrane integrity of the sensitive cells was disrupted by exposure to Scutellariae Radix extract as the $\beta$-galactosidase test on experimental substrate ONPG(o-nitrophenyl-$\beta$-D-galacto-pyranoside)

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The Effect of Fertilization-Promoting Peptide on Fertilizing Ability and Glycosidase Activity In Vitro of Frozen-Thawed Boar Spermatozoa

  • Hwang, I.S.;Park, C.K.;H.T. Cheong;B.K. Yang;Kim, C.I.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.43-43
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    • 2001
  • This study evaluated the effect of fertilization-promoting peptide (FPP) on fertilizing ability and glycosidase activity in vitro of spermatozoa frozen-thawed in pig, Using chlortetracycline fluorescence analysis, the various glycosidase analyses and the oocyte penetration test, we have obtained evidence that FPP can promote the fertilizing ability and glycosidase activity of frozen-thawed spermatozoa in vitro. When frozen-thawed spermatozoa was washed with different concentrations of FPP, there were significantly (P<0.05) more acrosome-reacted in medium with 100 nM than 0, 50, 200 and 400 nM. The penetration rates were also highest in medium containing with 100 nM FPP (P<0.05). On the other hand, the $\beta$-N-acetylglucosaminidase activity was at least twofold higher than other glycosidase. In same glycosidase, however, there were no difference in medium with different concentrations of FPP In another experiment, spermatozoa preincubated in medium with or without FPP for 0, 1, 2, 3 and 4 h were inseminated with oocytes matured in vitro. The percentages of spermatozoa that reached acrosome reaction were affected by preincubation and were higher in medium with that than without FPP. When oocytes were inseminated with spermatozoa preincubated in medium with and without FPP during the different periods, however, penetration rates were decreased with preincubation periods of spermatozoa. On the other hand, when the sperm-oocyte were cultured for 4, 8, 12, 16, 20 and 24 h, the penetration rates were higher in spermatozoa preincubated with that than without FPP and had a tendency to increase as time of culture periods. However, The activities of $\alpha$-fucosidase, $\alpha$ -mannosidase, $\beta$-galactosidase and N-acetyl- $\beta$-D-glucosaminidase were higher in medium with that than without FPP regardless of periods of sperm preincubation and sperm-oocyte culture. These results suggest that FPP may play a positive role in promoting of sperm function and glycosidase activity in vitro in pig.

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Differential Cytotoxicity of Penta-O-galloyl-β-D-glucose in Human Cancer and Normal Cell Lines of Various Origins (사람의 다양한 조직에서 기원하는 암세포 및 정상세포에 대한 penta-O-galloyl-β-D-glucose의 세포독성 효과)

  • Lee, Hyeon-Jeong;Kim, Min-Gyeong;Lee, Song-Yeong;Song, Min-Hyock;Kim, Yoon-Dong;Ha, Jeong-Sook;Jeong, Gie-Joon;Rho, Gyu-Jin;Jeon, Byeong-Gyun
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1320-1329
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    • 2016
  • The present study examined the cytotoxic effects of 1, 2, 3, 4, 6-penta-O-galloyl-${\beta}$-D-glucose (PGG), known as the pentahydroxy gallic acid ester of glucose, in the various human cancer cell lines (A-549, MDA-MB-231, U87-MG, MCF-7 and PANC-1), normal MRC-5 fetal fibroblasts, and dental papilla tissue- derived mesenchymal stem cells (DPSCs). Significantly (p<0.05) lower half maximal inhibitory concentration ($IC_{50}$) values were observed in the A-549 and MDA-MB-231 cells showing a high proliferation capacity, compared with other cancer and normal cell lines with a relatively low proliferation capacity. The population doubling time (PDT) was significantly (p<0.05) higher in the $10{\mu}M$ PGG-treated cell lines than those of untreated control cell lines. The present study demonstrated that the $IC_{50}$ value increases proportionally to the extending PDT. A high cell number with senescence-associated ${\beta}-galactosidase$ activity was also observed in the $10{\mu}M$ PGG-treated cells compared with those of untreated control cells. Moreover, the level of telomerase activity was significantly (p<0.05) decreased with $10{\mu}M$ PGG treatment, especially in A-549 and MDA-MB-231 cells showing a high proliferation capacity. Based on these observations, PGG could serve as a potent agent for cancer chemotherapy, as its treatment was more effective in cells with a high proliferation capacity.

Co-Infection of the Rat Central Nervous System with Genetically Engineered Strains of Pseudorabies Virus (유전자 조작된 Pseudorabies 바이러스에 의한 흰쥐 중추신경계의 이중감염)

  • Kim Jin-Sang;Kwon Young-Shil
    • The Journal of Korean Physical Therapy
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    • v.11 no.2
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    • pp.81-92
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    • 1999
  • 중추신경계의 미주신경동쪽핵(DMV)내 유사핵분열후 신경세포로 외래 유전자를 전달하는 매개체로서 pseudorabies 바이러스(PRV)의 유전자 조작기술은 흰쥐의 결장내로 PRV를 주입시킨 후 복잡한 신경로 추적에 관한 연구에서 하나의 바이러스에 의해 얻어지는 것보다 더욱 유용한 결과산출이 가능하게 하였다. 본 연구에서는 흰쥐의 생체내 실험모델로 하나의 바이러스 또는 이중 바이러스 주입에 PRV의 유전자 조작된 2종 바이러스를 사용하였다. 이 2종의 바이러스는 PRV의 Bartha 종에서 유래되었지만 면역조직화학적으로 검출할 수 있는 동일한 유전산물을 산출할 수 있도록 구성되었다. PRV-BaBlu는 PRV 게놈의 Us 구역 중 gC 자리에 lacZ 유전자를 삽입하여 산출되었는데 $\beta-galactosidase$ 발현은 이 바이러스에 감염된 신경원의 독특한 표시자로 나타났다. PRV-D는 2가지 단계에 의해 조성되었는데 첫째, PRV-Bartha의 Us 구역의 일부 유전자를 제거하고, 야생형인 PRV-Be DNA로 복구시켰는데 이로써 PRV-D는 PRV-Bartha 또는 PRV-Bablu에 존재하고 있지 않는 외피 당단백질인 gE와 gI를 지니게 되었다. 본 연구의 결과는 다음과 같았다. 첫째, PRV-D의 개별적 접종에 의해 얻어진 감염은 PRV-BaBlu에 의한 동일 신경회로의 감염보다 유의하게 빨랐다. 둘째, 유전자 조작된 PRV의 변이종은 변이종 상호간 및 부모 바이러스와 상이하였다. PRV-D는 PRV-Bartha 또는 PRV-BaBlu보다 감염독성이 더 강했고, PRV-BaBlu는 PRV-Bartha보다 감염독성이 약했다. 셋째, 결장을 지배하는 미주신경동쪽핵내 신경원은 변이종 바이러스들을 동시에 접종하였을 경우 이중감염을 나타내었다.

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Construction of Yeast Vectors Potentially Useful for Expression of Eukaryotic Genes as ${\beta}$-galactosidase Fusion Proteins

  • Chung, Kyung-Sook;Choi, Won-Ja;Lee, Hee-Won;Kim, Kyu-Won;Yoo, Hyang-Sook
    • BMB Reports
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    • v.29 no.4
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    • pp.359-364
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    • 1996
  • By both in vitro hydroxylamine mutagenesis of the wild type 3-phosphoglycerate kinase gene (PGK) promoter DNA and insertion of the leu2-d gene, we have created yeast expression vectors potentially useful for production of eukaryotic genes in yeast. The guanine (G) to adenine (A) change at the -3 position from the ATG start codon of the PGK promoter-based vector rendered a 6~7 times elevated expression of the adjacent eukaryotic gene, and insertion of the leu2-d gene in the vector containing the mutated PGK promoter further enhanced the expression of the gene. When expression of the AIDS virus HIV1-gagP17 gene in a lacZ fusion form was examined with this new vector, a 15 times higher level of expression than that from the original PGK promoter was observed. Northern and Southern analysis showed that this elevated expression is due to the production of a high copy number of mRNA by leu2-d gene functioning and by efficient translation of the produced mRNA. Thus, the vector that contained the A at the -3 position from the ATG start codon in the promoter region and the leu2-d gene shows increased expression capability and will be potentially useful for production of eukaryotic genes in yeast.

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Migration Activity of Chicken Gonadal Primordial Germ Cells (gPGCs) and Post-transfer Localization of LacZ-transfected gPGCs in the Embryonic Gonads

  • Jeong, D.K.;Han, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.9
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    • pp.1227-1231
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    • 2002
  • A powerful tool for chicken transgenesis could be established by employing a germline chimera production through primordial germ cell transplantation. This study was conducted to examine whether foreign gene-transfected gonadal primordial germ cells (gPGCs) have a migration activity into the gonad after transfer to recipient embryos. In Experiment 1, gPGCs of Korean Ogol Chicken were retrieved from 5.5-day-old embryos and subsequently transferred to the dorsal aorta of 2.5-day-old White Leghorn embryos after being labeled with PKH26 fluorescent dye. To confirm migration activity after transplantation, recipient embryos were sacrificed and examined on 3 days after transfer. Sex determination was concomitantly undertaken to examine whether sex of recipient embryos could affect the migration activity of gPGCs. All of embryonic gonads examined showed positive signals with PKH26 fluorescence and W-chromosome specific band by polymerase chain reaction (PCR) was detected in male embryos when gPGCs with ZW chromosome were transferred to recipient embryos. In Experiment 2, retrieved gPGCs were transfected with LacZ gene-containing cytomegalovirus promoter ($pCMV{\beta}$) by electroporation and subsequently transferred to recipient embryos. LacZ gene expression was identified in the gonads of 6 or 10-day-old recipient embryos and hatched-chicks. A total of 20 embryos and 12 hatched-chicks were examined and 11 of them (10 embryos and one hatched chicken; 11/32=34.4%) expressed $\beta$-galactosidase, a marker substance of LacZ gene. The results of this study demonstrated that foreign gene-transfected gPGCs can migrate and settle down into the gonad after being transferred into the blood vessel of the recipient embryos. This established technique will contribute to developing a peer biotechnology for transgenic chicken.