• Title/Summary/Keyword: Culture medium volume

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Isolation of Phytase Producing Pseudomonas fragi and Optimization of its Phytase Production (Acid Phytase를 생산하는 Pseudomonas fragi의 분리와 phytase의 생산조건)

  • Kim, Young-Jin;Jang, Eun-Seok;In, Man-Jin;Oh, Nam-Soon
    • Applied Biological Chemistry
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    • v.46 no.4
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    • pp.291-298
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    • 2003
  • A bacterial strain producing a high level of an extracellular phytase was isolated from livestock waste water, identified as a strain of Pseudomonas fragi and designated as Pseudomonas fragi Y9451. Under the phytase production medium, the activity of phytase reached the highest level after 120 hours of incubation. On the effect of carbon sources on the phytase production, the most favorable carbon source for phytase production was fructose. As for the effect of nitrogen sources, high levels of phytase activity were detected in the medium containing nutrient broth as the nitrogen source. Free $PO_4^{3-}$ inhibited phytase production with increasing concentration of $KE_2PO_4$ and phytate in the media. The addition of $CaCl_2$ and $MgSO_4$ also resulted in the inhibition of phytase production. To investigate the effect of aeration on the phytase production, different volumes of culture broth in Erlenmeyer flasks were incubated in rotary shaker at the speed of 200 rpm. As a result, a high level of phytase activity was detected at small volume of culture broth as compared to larger volume because of its more aerobic condition.

Effect of Lactic Acid Bacteria on the Qualities of White Pan Bread (빵의 품질에 미치는 유산균의 영향)

  • 장준형;안재법
    • The Korean Journal of Food And Nutrition
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    • v.9 no.4
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    • pp.509-515
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    • 1996
  • The effects of sour liquid ferments with lactic acid bacteria on the baking properties and qualities of White Pan Bread were studied. The mixed culture of Lactobacillus brevis and Lactobacillus plantarum had higher acid equivalents and lower pH-values than single or mixed culture of other lactic acid bacteria which had been used for traditional sour dough bread. Optimum conditions of the incubation of lactic acid bacteria, which are incubation temperature time and culture medium compositions for lactic fermentation, were also investigated to find out optimum activity for good bread making. The mixed culture of L. brevis and l. plantarum incubated for 24 hours at 3$0^{\circ}C$ had the most optimum activity for bread manufacturing process and the qualities of the products. The addition of sour liquid ferments to the sponge dough effected on fermentation activity of the sponge dough to lower the level of pH to 4.64 and to produce more total titratable acidity(TTA) of 0.545, whereas conventional sponge dough bread had 0.46% of TTA. On comparison with control bread, the bread made with sour liquid ferments was found to have better specific volume, taste, symmetry, especially, organoleptic characteristics due to lactic acid, acetic acid and amino acid produced by lactic acid bacteria. Sour dough bread with liquid ferment was considered to be more effective to the inhibition of staling during storage for 6 days at $25^{\circ}C$ and to have longer shelf-than control.

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Development of In Vitro Porcine Oocytes Following Intracytoplasmic Injection of Sperm-Mediated GFP Gene

  • Kim, J.H.;Seong, H.H.;Park, J.K.;Im, S.K.;Kim, S.W.;Lee, Y.K.;Lee, P.Y.;Choi, Y.J.;Kim, Y.K.;Kim, J.H.;Chang, W.K.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.69-69
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    • 2002
  • Transgenic animals production tools have been valuable for research and purpose. The current methods of gene transfer, microinjection and nuclear transfer, which are widely used in transgenic animal production, but all most methods has only had limited success in production of larger species. Here, we report the possibility of a sperm-mediated gene transfer method in porcine embryos. Oocytes were collected from ovaries harvested at a local slaughterhouse were matured in 500${mu}ell$ drops of TCM-199 under mineral oil at 38.5$^{\circ}C$ in a humidified atmosphere of 5%CO2 in air. After 42-43h of in vitro maturation oocytes were denuded. for sperm injection into the cytoplasm of the porcine oocytes, sperm suspension in NIM medium are subjected extraction with TritonX-100 before mixing with a green fluorescent gene (GFP). Sperm with Tritonx-100 were prepared by adding TritonX-100 to a final volume of 0.05% in the sperm suspension and mixing by trituration for 60s before two wishes in NIM medium at 2$^{\circ}C$. A(ter wishing, sperm were mixed with TritonX-100 at $25^{\circ}C$ followed by washes at 2$^{\circ}C$. Sperm were resuspended in ice cold NIM to a final volume of 400${mu}ell$ and 2-20ng/${mu}ell$ DNA were triturated on ice for 60s. All microinjection was performed in HEPES-buffered CZB medium at room temperature within 2h. After culture in NCSU-23 for 72h, percent of porcine embryos transfected GFP gene are 20.7%(6/29) in 20ng/${mu}ell$ sperm-DNA mixed group and other groups were 3.7 %(2/54)and 4.7%(3/67). These data suggests that sperm-mediated gene transfer method should be used to the production tool of transgenic pig efficiently.

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DNA Delivery into Embryogenic Cells of Zoysiagrass(Zoysia japonica Steud.) and Rice(Oryza sativa L.) by Electroporation (Electroporation을 이용한 잔디(Zoysia japonica Steud.) 및 벼(Oryza sativa L.) 배발생세포로의 DNA 도입)

  • 박건환;최준수;윤충호;안병준
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.5
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    • pp.309-314
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    • 1994
  • To develop simple and efficient transformation methods of monocotyledonous plane, electroporation-mediated delivery of DNA into intact embryogenic cell clumps was investigated in zoysiagrass and rice. Calli of zoysiagrass, induced from 3-week-old immature embryos, were suspension-cultured in MS basic medium supplemented with 1.0 mg/t 2.4-D and used for elechuporation. Calli, derived from immature inflorescences of 20 mm lenth of rice, were also suspension-cultured on N6 basic medium supplemented with 1.0 mg/L 2.4-D. Suspension-cultured embryogenic cell clumps were electroporated in liqid MS medium added with a Plasmid DNA (30 $\mu\textrm{m}$/ml), pGA1074, encoding ${\beta}$-glucuronidiase (GUS). DNA delivery into the cells through cell walls and cell membrane was confirmed by the transient expression of the GUS gene. Cell clumps of zoysiagrass and rice, electroporated with 400 volt at 800 pF capacitance, expressed GUS gene activity at a mean frequency of 25 units (one unit = one clony of blue cells) per 200 ${\mu}\ell$ of packed cell volume. Untreated cells and healed non-embryogenic cells did not exhibit GUS activity These results indicate that electroporation-mediated transformation can use intact embryogenic cells (thus avoiding the use protoplasts) in zoysiagrass and rice.

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Cultural characteristics and fruiting body productivity of new Lentinula edodes cultivar 'Jadam' in rod-type sawdust cultivation (표고 봉형 톱밥재배시 배양중 침공처리가 '자담'의 발이 및 자실체 생산에 미치는 영향)

  • Jeong-Han Kim;Young-Ju Kang;Yeon-Jin Kim;Chae-Young Lee;Jun-Yeong Choi;Chan-Jung Lee;Gab-June Lim
    • Journal of Mushroom
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    • v.21 no.3
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    • pp.160-166
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    • 2023
  • The effects of punching treatment on mycelial culture and fruiting body productivity were investigated in a new Lentinula edodes cultivar, "Jadam", in sawdust medium for the stable production of oak mushroom. As the punching volume and number increased, the weight loss rate and color difference increased and the L value decreased. After spawn inoculation, the sawdust medium temperature and CO2 concentration reached their highest values at 33 and 19 days of incubation, respectively. The O2 concentration showed the lowest value on the 14th day of incubation, which was the opposite pattern to the CO2 concentration. As the punching volume and the number increased, the medium temperature and O2 concentration increased, and the CO2 concentration decreased. Higher punching volumes and numbers resulted in higher temperatures and lower CO2 concentrations. The best fruiting body yield was 5 × 70 mm - 30 (punching diameter × depth - number), and the total yield after three cycles was 644.7 g.

Establishment of Optimal Production Conditions of Transglucosidase Produced by Aspergillus niger (Aspergillus niger가 생산하는 transglucosidase의 최적 생산 조건 확립)

  • Lee, Jun-Yeob;Gang, Seongho;Kim, Jong-Sik;Chung, Chungwook
    • Journal of Life Science
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    • v.28 no.8
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    • pp.969-976
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    • 2018
  • In this study, transglucosidase (TG), an enzyme produced by Aspergillus niger, synthesized isomaltooligosaccharide from ${\alpha}-(1{\rightarrow}4)$ linked substrates. The highest TG-producing A. niger KCTC6913 was selected from six kinds of species, and optimized TG producing conditions were established. Five different carbon sources (potato starch, sweet potato starch, corn starch, wheat starch, and dextrin) and three different nitrogen sources (yeast extract, malt extract, and beef extract) were tested to establish the carbon and nitrogen sources favorable for TG production. Measurements of TG activity after an initial culture at pH 5.0 for 15 days revealed that potato starch and yeast extract, which are basic culture media, resulted in the highest TG activity. In addition, A. niger KCTC6913 increased TG production under aerobic conditions and a controlled carbon/nitrogen ratio. In conclusion, to evaluate TG activity in the established optimal medium, it is confirmed that the basal and potato dextrose broth medium were used as a control, and the highest TG production was measured, which was highlighted in the established optimal medium.

The Effect of Co-existing Artemia sp. on the Rotifer Brachionus rotundigformis Population Growth (Rotife 배양조에 혼재된 Artemia가 Rotifer 의 증식에 미치는 영향)

  • Jung, Min-Min;Rho, Sum;Kim, Pil-Yun
    • Journal of Aquaculture
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    • v.11 no.1
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    • pp.99-103
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    • 1998
  • Artemia often observed as a co-existing organism in the mass culture tank of marine rotifer. The rotifer and Atremia are commonly used as food organisms in the marine fish larvae rearing. In this study, interspecific relation between the rotifer Brachionus rotundiformis (formely called S-type) and anostracan Artemia of the two developmental stages (0 and 19 day old after hatching) were investigated in the larboratory. The population growth of B. rotundiformis and one of the stage (nauplius or adult) of Artemia in mixed culture was compared with that of each single species culture. Culture period was 16 days. Every two days, the number of organisms in each species was counted and transferred to a fresh medium containing $7{\times}10^5$cells/ml of food Nannochloropsis oculata. Culture volume, temperature, salinity and photoperiod were set at 40ml, $25^{\circ}C.$, 22ppt and 24h all dark except to observation time, respectively. The rotifer population growth was greatly decreased by co-existence with Artemia. The coexisting Artemia suppressed the rotifer population growth due to it's high filtering speed for food (N. oculata). This study suggested that contamination by Artemia must be prevented for the stable rotifer production in the rotifer mass culture tank.

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Immune Enhancement of Polysaccharide from Submerged Culture with Phellinus linteus in the Medium Supplemented with Ginseng Extract (수삼추출물 첨가 혼합배지에서 조제된 상황 균사체 심부배양물 다당획분의 면역활성 증진)

  • Kim, Hoon;Song, Ki-Yun;Jeong, Jae-Hyun;Jeong, Heon-Sang;Lee, Hyeon-Yong;Yu, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.20-28
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    • 2011
  • Crude polysaccharide (CP) was fractionated from the submerged culture (containing both mycelia and culture broth, SC) with Phellinus linteus (PL) in mushroom complete medium (MCM) supplemented with ginseng extract ($65^{\circ}$Bx, GE) to enhance the immune activity. PL-GE-15-CP from SC cultivated in MCM supplemented with GE-15% (v/v, a ratio of MCM volume to GE) showed significantly higher macrophage stimulation (1.45 fold of the saline control at $100{\mu}g$/mL) than PL-GE-5 and 10-CP with GE-5 and 10%, or PL-CP from SC without GE. The potent intestinal immune system modulating activity through Peyer's patch was also obtained by PL-GE-15-CP (1.46 fold). When PL-GE-15-CP further fractionated on DEAE-Sepharose CL-6B (Cl- form), PL-GE-15-CP-II was the significantly higher than others from PL-GE-15-CP or PL-CP on macrophage stimulation, interleukin (IL)-12 production and intestinal immune system modulation (1.54, 3.96 and 1.56 fold, respectively). PL-GE-15-CP-II also had higher anti-metastatic activity against colon 26-M3.1 carcinoma cell (57.3% inhibition of tumor control, $200{\mu}g$/mouse) rather than PL-CP-II. This active fraction (PL-GE-15-CP-II) mainly contained neutral sugar (82.45%) and uronic acid (12.99%), and component sugar analysis showed that PL-GE-15-CP-II consisted mainly of uronic acid, Ara, Man, Gal and Glc (molar ratio of 0.52:0.97:0.63:1.00:0.54). Furthermore, the activity of GE culture was higher compared with culture without GE, indicating that GE helped to enhance the immune activity of P. linteus; also, it is assumed that the polysaccharide plays an important role in immune enhancement.

Glycogen Metabolism in Vibrio vulnificus Affected by malP and malQ

  • Han, Ah-Reum;Lee, Yeon-Ju;Wang, Tianshi;Kim, Jung-Wan
    • Microbiology and Biotechnology Letters
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    • v.46 no.1
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    • pp.29-39
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    • 2018
  • Vibrio vulnificus needs various responsive mechanisms to survive and transmit successfully in alternative niches of human and marine environments, and to ensure the acquisition of steady energy supply to facilitate such unique life style. The bacterium had genetic constitution very different from that of Escherichia coli regarding metabolism of glycogen, a major energy reserve. V. vulnificus accumulated more glycogen than other bacteria and at various levels according to culture medium and carbon source supplied in excess. Glycogen was accumulated to the highest level in Luria-Bertani (3.08 mg/mg protein) and heart infusion (4.30 mg/mg protein) complex media supplemented with 1% (w/v) maltodextrin at 3 h into the stationary phase. Regarding effect of carbon source, more glycogen was accumulated when maltodextrin (2.34 mg/mg protein) was added than when glucose or maltose (0.78.1-14 mg/mg protein) was added as an excessive carbon source to M9 minimal medium, suggesting that maltodextrin metabolism might affect glycogen metabolism very closely. These results were supported by the analysis using the malP (encoding a maltodextrin phosphorylase) and malQ (encoding a 4-${\alpha}$-glucanotransferase) mutants, which accumulated much less glycogen than wild type when either glucose or maltodextrin was supplied as an excessive carbon source, but at different levels (3.1-80.3% of wild type glycogen). Therefore, multiple pathways for glycogen metabolism were likely to function in V. vulnificus and that responding to maltodextrin might be more efficient in synthesizing glycogen. All of the glycogen samples from 3 V. vulnificus strains under various conditions showed a narrow side chain length distribution with short chains (G4-G6) as major ones. Not only the comparatively large accumulation volume but also the structure of glycogen in V. vulnificus, compared to other bacteria, may explain durability of the bacterium in external environment.

Growth and Morphological Changes in Scenedesmus dimorphus Induced by Substances Released from Grazers, Daphnia magna and Moina macrocopa (동물플랑크톤 Daphnia magna와 Moina macrocopa에서 유도된 분비물질에 의한 Scenedesmus dimorphus의 형태변화)

  • Jang, Min-Ho;Joo, Gae-Jae;Takamura, Noriko;Ha, Kyong
    • Korean Journal of Ecology and Environment
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    • v.34 no.4 s.96
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    • pp.285-291
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    • 2001
  • Grazer-induced colony formation was examined using a green alga Scenedesmus dimorphus (T$\ddot{u}$rpin) K$\ddot{u}$tzing. Algae were cultured in a medium with or without filtered water taken from cultures of Daphnia magna Straus (300 ind./L) or Moina macrocopa Straus (500 ind./L). The exposure to zooplankton filtered water (ZFW)promoted colony formation in S. dimorphus, with the magnitude of this response being directly proportional to the relative volume of ZFW that was added to the culture medium. The number of cells/colony and mean particle biovolume of S. dimorphus increased between 24 and 72 hours after exposure to ZFW, most likely due to the influence of chemicals released from D. magna or M. macrocopa, and possily as a defense mechanism against zooplankton grazing.

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