• Title/Summary/Keyword: liquid culture

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Studies on the survival Rate after Slow and Ultrarapid Frozen-Thawing of Porcine Embryos (돼지 수정란의 완만 및 초급속 동결 융해후의 생존성에 관한 연구)

  • 이봉구;김상근;이규승
    • Korean Journal of Animal Reproduction
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    • v.16 no.2
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    • pp.117-123
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    • 1992
  • This Study was carried out ot investigate the effects of concentration and equilibration time of cryoprotective aagents on survival rate of slowly and ultrarapidly frozen porcine embryos. The porcine embryos following dehydration by cryoprotective agents and 0.25M sucrose were slowly freezed(from 2$0^{\circ}C$ to -7$^{\circ}C$/-1$^{\circ}C$/min., from -7$^{\circ}C$ to -35$^{\circ}C$/-0.2$^{\circ}C$/min., from -35$^{\circ}C$ to -38$^{\circ}C$/-0.3$^{\circ}C$/min.) by Cell Freezer and directly plunged into liquid nitrogen and thawed in 38$^{\circ}C$ water bath. Survival rate was defined as development rate to the morula and blastocyst stage after in vitro culture or by FDA test. The results are summarized as follows : 1. The survival rates of porcine embryos after slow frozen-thawing in the freezing medium of 0.25M sucrose added 2.0M glycerol, 3.0M DMSO, 2.0M propanediol or 2.0M glycerol+2.0M propanediol was 80.6, 84.7, 75.0 or 78.8%, respectively. 2. The survival rates of porcine embryos after slow frozen-thawing in the freezing medium of 0.50M sucrose added 2.0M glycerol, 3.0M DMSO, 2.0M propanediol or 2.0M glycerol+2.0M propanediol was 80.9, 82.4, 73.1 or 77.1%, respectively. 3. The survival rates of porcine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucroese added 2.0M glycerol, 3.0M DMSO, 2.0M propanediol or 2.0M glycerol+2.0M propanediol was 65.3, 68.6, 63.2 or 59.9%, respectively. 4. The survival rates of porcine embryos after ultrapid frozen-thawing in the freezing medium of 0.50M sucrose added 2.0M glycerol, 3.0M DMSO, 2.0 propanediol or 2.0M glycerol+2.0M propanediol was 67.5, 62.9, 56.9, or 62.8%, respectively. 5. The higher survival rate of porcine embryos was attained at the short period ofequilibration time(5min.) in the freezing medium added 0.25M sucrose and 3.0 DMSO compared to those of 10 or 20min. equilibration time in the same condition.

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Studies on the Survival Rates after Ultrarapidly Frozen-Thawing of Porcine Embryos (돼지 수정난의 초급속 동결 융해후의 생존성에 관한 연구)

  • 김상근;이봉구
    • Korean Journal of Animal Reproduction
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    • v.16 no.2
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    • pp.125-131
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    • 1992
  • This study was carried out to investigate the effects of concentration and equilibration time of cryoprotective agents on the survival rate of slowly and ultrarapidly frozen porcine embryos. The porcine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 38$^{\circ}C$ water bath. Survival rate was defined as development rat to the morula and blastocyst stage after in vitro culture or by FDA test. The results are summarized as follows : 1. The survival rates of porcine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0, 2.5, 3.0, 3.5, or 4.0M glycerol was 65.3, 61.8, 64.3, 59.4 or 39.4%, respectively. Addition of 0.25M sucrose into the freezing medium containing 2.0M glycerol showed higher survival rate than those of 2.5~4.0M glycerol. 2. The survival rates of porcine embryos after ultraradpid frozen-thawing in the freezing medium of 0.25M sucroese added 2.0, 2.5, 3.0, 3.5 or 4.0M DMSO was 65.6, 67.6, 68.6, 60.6 or 23.6%, respectively. However, addition of 0.25M sucrose into the freezing medium containing 3.0M DMSO showed higher survival rate than those of 2.0, 2.5, 3.5 or 4.0M DMSO. 3. The survival rates of porcine embryos after ultrapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0, 2.5, 3.0, 3.5 or 4.0M propanediol was 63.2, 60.3, 62.1, 52.3 or 24.3%, respectively. Addition of 0.25M sucroese into the freezing medium containing 2.0M propanediol showed higher survival rate than those of 2.5~4.0M glycerol. 4. The survival rates of porcine embryos after ultrarapid frozen-thawing the freezing medium of 2.0M glycerol added 0.10, 0.25, 0.50 or 0.75M sucrose was 61.8, 70.8, 67.6 or 52.2%, respectively. Addition of 2.0M glycerol into the freezing medium containing 0.25M sucreose showed higher survival rate than that those of 0.10, 0.50 or 0.75M sucrose. 5. The higher suvival rate of porcine embryos were attained at short period of equilibration time 92.5~5min.) in the freezing medium added 0.25M sucreose and 3.0M compared to those of 10 or 20min. equilibration time in the same condition.

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The Effects of Mashing and Maturing Conditions on The Quality of Korean Traditional Kanjang(Soysauce) (한국전통간장의 품질에 미치는 사입과 숙성조건의 영향)

  • Choi, Jong-Dong;Im, Moo-Hyeog;Chung, Hyun-Chae;Lee, Coon-Woo;Kim, Young-Ho;Choi, Cheong;Choi, Kwang-Soo
    • Applied Biological Chemistry
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    • v.40 no.5
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    • pp.365-368
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    • 1997
  • This study was carried out to investigate the optimum mashing and maturing conditions for Korean traditional Kanjang(soy sauce) production and to reduce the fermentation period. The effects of maturing time of soy sauce mash, maturing temperature, salt concentration and the ratio of Meju to salt brine on the quality of Kaniang(total nitrogen, pH and color) were examined. Soy sauce pigments and about 90% of N constituents contained in soybean Meju(Koji) in soy sauce mash were degraded and solubilized into liquid portion (soy sauce) of the mash within five days of maturing at $30^{\circ}C$ with the mashing ratio(weight/volume) of 1 : 4 of soybean(as raw soybean) to 20% salt brine. No remarkable effects of soy sauce maturing temperature in the range of $5^{\circ}C{\sim}30^{\circ}$ on the digestion and solubilization of N components and pigment extraction during five days of soy sauce mash maturing were observed. Optimum mashing salt brine concentration for the digestion and solubilization of N components and pigment extraction during soy sauce maturing at $30^{\circ}C$ were observed to be in the range of $15{\sim}20%$. The suitable mashing ratio of Meju to salt brine (wt./vol.) to match N content of the standards of identity of Korean traditional Kanjang(soy sauce) was found to be below 1 : 5.

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Phosphate Uptake by Acinetobacter lwoffi PO8 and Accumulation (Acinetobacter lwoffi PO8에 의한 인산흡수 및 축적)

  • Yoon, Min-Ho;Ko, Jung-Youn;Choi, Woo-Young;Shin, Kong-Sik
    • Applied Biological Chemistry
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    • v.43 no.3
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    • pp.163-168
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    • 2000
  • To remove phosphate accumulated in the soil and water, Acinetobacter lwoffi PO8 possessing a high ability to accumulate phosphate was isolated from a active sludge. Bacterium was cultured in the liquid medium containing $150\;{\mu}g/mL$ of phosphate at $30^{\circ}C$ in different culture conditions to examine intracellular phosphate uptake. The initial pH in the range of $7.5{\sim}8.5$ was effective on the growth and phosphate uptake of the strain. Glycerol and arabinose used as a carbon sources showed 93 and 91% the phsphate uptake, respectively. Among the nitrogen sources, ammonium salt such as $NH_4NO_3$ and $(NH_4)_2SO_4$ was effectively utilized on the phosphate uptake compared with amino compounds. The rate of phosphate uptake of $NH_4NO_3$, and $(NH_4)_2SO_4$, was 95 and 96%, respectively The growth and Phosphate uptake ability in the strain were significantly promoted when metal ions were added in the medium; $Co^{2+}$, however, was not utilized by the strain. The capacity of phosphate uptake was enhanced to $10{\sim}20%$ when arginine, methionine, or lysine was added. Using $^{32}P$ to examine the uptake Pattern of intracellular phosphate, experiment result showed that polyphosphate was largely found in the fraction of intracellular inorganic phosphate of Acinetobacter lwoffi PO8.

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Purification and Characterization of Urushiol Induced Laccase Isoenzyme from Fomitella fraxinea (Urushiol에 의해 유도된 장수버섯 laccase isoenzyme의 정제 및 특성)

  • Choi, Han-Seok;Park, Hyo-Suk;Yeo, Soo-Hwan;Jeong, Seok-Tae;Choi, Ji-Ho;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.152-159
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    • 2010
  • The influence of urushiol, as an allergen on laccase property of Fomitella fraxinea was investigated. The enzyme production was reached to the highest level after 10 days, cultivation and the activity and mycelial biomass were increased by 2.5 and 1.5 folds, respectively, by adding urushiol in the culture medium. In liquid cultures using a Cu Mn-free medium, laccase lactivity was decreased by 3.8-9.2 folds, with similar dry cell weight. Two isoenzymes, were purified using anion exchange, hydrophobic interaction and size-exclusion chromatographies. Both isoenzymes are monomeric proteins, with $M_W$ around 67 kDa(Lac1) and 66 kDa(Lac2), and isoelectric points of 3.67 and 3.81. The optimal conditions for purified isoenzymes were found to be pH 4.5-5.0 and $30-35^{\circ}C$. Activity decreased by the addition of $Fe^{2+}$, $Mg^{2+}$, $Na^+$, and strongly inhibited by EDTA and sodium azide.

Screening for Fermentative Microorganisms that Grow on Brown Rice with High Amylase and Pretense Activities (Amylase와 Protease의 활성이 높은 현미 발효 미생물의 선별)

  • Kim Ki-Yeon;Kim Hee-Gyu;Song Byeong-Chul;Cha Chang-Jun
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.160-163
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    • 2006
  • Brown rice contains rice bran and germ with higher nutritional value and dietary fiber content compared with the polished rice. However, brown rice has a limitation of poor digestion. fermented brown rice could be better nutritional source and improve digestibility. Therefore, we tried to select good fermentative microorganisms which have nutritional values with high amylase and protease activities, and probiotic effects. Nineteen micro-organisms, including eight Bacillus strains isolated from Chongkukjang and 11 lactic acid bacteria, were screened for the fermentation ability and enzyme production. The liquid broths containing 2.5%(w/v) of raw brown rice powder as a sole nutritional source were used for culture media. Among the strains tested, all of the Bacillus strains and two lactic acid bacteria (Leuconostoc gelidum and Pediococcus pentosaceus) showed increase in cell population and enzyme activities. The viable cell counts of all the Bacillus strains and two lactic acid bacteria exceeded $10^7 CFU/mL$. The maximal enzyme activities produced by Bacillus sp. Bl, Bacillus sp. B2, Bacillus sp. B11, L. gelidum and P. pentosaceus were 17.85, 17.50, 17.10, 17.10 and 3.24 U/mL for amylase and 22.48, 22.04, 23.76, 12.13, and 3.4 U/mL for pretense, respectively. Therefore, the results of this study demonstrated that the above strains could be potential starters for the fermentation of raw brown rice.

Extracellular Polysaccharide Produced by a New Methylotrophic Isolate (새로운 메탄올 자화세균이 생산하는 세포외 다당류)

  • Lee, Ho J.;Kim, Si W.;Kim, Young M.
    • Korean Journal of Microbiology
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    • v.34 no.4
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    • pp.212-218
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    • 1998
  • An obligately methylotrophic bacterium which produces extracellular polysaccharide (EPS) was isolated through methanol-enrichment culture technique. The isolate was aerobic, nonmotile, and gram negative rod and exibited catalase, but no oxidase, activity. Plasmid, carotenoid, and poly-${\beta}$-hydroxybutyric acid were not found. The guanine plus cytosine content of DNA was 52-56%. The isolate was found to grow only on methanol and monomethylamine. Growth was optimal ($t_d=2.4h$) at $35^{\circ}C$ and pH 6.5 in a mineral medium containing 0.5% (v/v) methanol, 25 mM phosphate, and 0.212% ammonium sulfate. Methanol was assimilated through the ribulose monophosphate pathway. Maximun amount of EPS was produced in cells growing at the mid-stationary growth phase at $30^{\circ}C$ in a mineral medium (PH 6.5) containing 1.0% (v/v) methanol in the CIN ratio of 54.7. Thin-layer chromatographic and high performance liquid chromatographic analysis revealed that the EPS was composed of glucose and galactose. EPS which was not treated with ethanol (Pbe) exhibited stable viscosity under various concentrations of salts and temperatures hut showed high viscosity at low pH. EPS precipitated with ethanol (Pae) was found to be more stable in viscosity than the Pbe at various salt concentrations, temperatures, and pH. The Pae also exhibited higher viscosity than the Pbe and xanthan gum. Scanning electron microscopy revealed that the lyophilized Pbe and Pae have a multi-layered structure and a structure of thick fibers, respectively.

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Effects of Excising In Vitro-Formed Roots on Acclimatization of Micropropagated Cassava Plantlets (카사바의 미세증식에서 기내 발생 부정근의 절단이 순화에 미치는 영향)

  • Yoon, Sil;Cho, Duck-Yee;Soh, Woong Young
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.103-108
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    • 2001
  • The in vitro plantlets of cassava (Manihot esculenta Crantz cv. MColl 22) could be regenerated from nodal explant cultures in a liquid MS basal medium containing 0.01 mg/L zeatin for 2 weeks. The plantlets of 1.5∼2.5 cm in shoot length were transplanted to a glass bottle containing fine sand and acclimated under non-sterile conditions after excising their intact roots by: 1) prune leaving roots base of 1∼1.5 cm; 2) complete removal of roots; and 3) cutting off the rooting zone. The majority of in vitro-formed intact roots continued growth after transferred to soil, and all of the damaged roots stopped further growth. The plantlets with excised roots began to develop new roots within 7∼10 days after being transferred to a glass bottle, and a few of the pruned roots developed lateral roots from the remaining portion. Pruning and removal of in vitro roots resulted in a high survival rate (>87%), and did not significantly affect ex vitro root regeneration and acclimation, but the plantlets in which the rooting zone had been cut-off showed 73% survival rate. Pruning or removal of in vitro roots before transfer of plantlets is recommended for useful method of commercial micropropagation because of easier handling and high survival rate of plantlets.

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Establishment of Optimal Fermentation Conditions for Steam-dried Ginseng Berry via Friendly Bacteria and Its Antioxidant Activities (생체친화성 균주에 의한 인삼열매증포 추출물의 최적발효조건 및 항산화활성)

  • Kim, Seung Tae;Kim, Hee Jung;Jang, Su Kil;Lee, Do Ik;Joo, Seong Soo
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.77-83
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    • 2013
  • In this study, we observed optimal conditions and suitable bacteria for the fermentation of steam-dried ginseng berry extracts (SGB) and determined antioxidant effects of the fermented extracts. Five bacteria (Lactobacillus fermentarum, L. plantarum, L. brevis, L. casei, Bacillus subtillis) were examined on their growth activities and viabilities in various culture temperatures ($25-35^{\circ}C$) and concentrations (25-100%). L. plantarum was considered to be the most suitable bacteria for the fermentation in both growth activity and viability. Moreover, the extracts fermented with L. plantarum showed more potent antioxidant efficacy in both 1,1-diphenyl-2-picrylhydrazyl radical and hydroxyl radical scavenging assay. High performance liquid chromatography analysis revealed that fermentation with L. plantarum changed the contents and components of ginsenosides. In conclusion, these data suggest that L. plantarum efficiently ferment SGB and the fermented extracts may have therapeutical values against oxidative stress and be a good candidate in adjuvant therapy where ginsenoside would be the main composition.

Isolation of an Agarase-producing Persicobacter sp. DH-3 and Characterization of its β-agarase (Agarase를 생산하는 Persicobacter sp. DH-3의 분리 및 β-agarase의 특성)

  • Heo, Da-Hye;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.29 no.2
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    • pp.158-163
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    • 2019
  • The purpose of this study was to isolate a new marine agarase-producing bacterium. Agarase can hydrolyze agar and agarose to produce agarooligosaccharides or neoagarooligosaccharides, which possess many physiological functions. Strain DH-3 was isolated from seawater collected from the coast of Yeosu at Jeollanam province, Korea. A 16S rDNA sequence analysis showed this strain to be Persicobacter sp. DH-3. Extracellular agarase was prepared from culture media of Persicobacter sp. DH-3 and used for characterization. Relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 50, 55, 70, 100, 90, and 50%, respectively. Relative activities at pH 5, 6, 7, and 8 were 75, 100, 90, and 75%, respectively. The enzyme showed maximum activity at $50^{\circ}C$ in a 20 mM Tris-HCl buffer at pH 6. This enzyme could be useful, as agar is in liquid state at $50^{\circ}C$. Agarase activities were maintained at 80% or more for 2 hr at 20, 30, and $40^{\circ}C$. Thin layer chromatography analysis suggested that Persicobacter sp. DH-3 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarohexaose and neoagarotetraose. In addition, zymogram analysis confirmed that Persicobacter sp. DH-3 produces at least three agar-degrading enzymes with molecular weights of 45, 70, and 140 kDa. Therefore, it is expected that agarases from Persicobacter sp. DH-3 could be used to produce functional neoagarooligosaccharides.