• Title/Summary/Keyword: chemically defined medium

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Production of Extracellular Protein from Bacillus sp. (Bacillus sp.에 의한 균체외 단백질의 생산)

  • Lee, Jae-Sook;Kim, Chan-Jo;Lee, Jong-Soo
    • Applied Biological Chemistry
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    • v.31 no.2
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    • pp.187-192
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    • 1988
  • Extracellular protein was produced from bacterium, which was classified as a strain belong to the genus Bacillus. This bacterium produced 1.4mg/ml of extracellular protein when grown on chemically defined medium containing 3.0% of glucose, 1.3% of urea and 2.5% of calcium carbonate at $30^{\circ}C$ for 48 hours. The addition of $250{\mu}g/ml$ of cephalexin was effective for it and was produced 2.0mg/ml of extracellular protein for 96 hours.

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Effects of Osmolarity and Vitamins on the In Vitro Development of Bovine Embryos in a Chemically Defined, Protein-free Culture Medium (무단백 한정배양액에서 삼투압 및 비타민이 소 수정란의 체외발생에 미치는 영향)

  • 김종홍;이상찬;김병기
    • Korean Journal of Animal Reproduction
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    • v.20 no.1
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    • pp.77-84
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    • 1996
  • The purpose of this study was to evaluate effects of osmolarity and vitamins on the in vitro development of embryos. Bovine embryo that had been matured and fertilized in vitro were cultured in simple, chemically defined, protein free medium (mTLP-PVA). When the osmolarity of the medium supplemented 0.35 mM phos-phate and 19 amino acid was changed by NaCI concentration, significantly(p<0.05) higher pro-portion of 1-cell embryos in the medium of 265 or 290 mOsm developed to the morula (32 ~ 35%) and blastocyst(24 ~ 28%) stage. When embryos were transferred to fresh medium containing 5.56mM glucose at 120hrs post-insemination, the highest proportion of embryos developed to mor-ula(40%) and blastocyst(32%) stages at 290 mOsm(p<0.05), although the value in morulae was not significantly different with that(35%) at 315 mOsm. Vitamins in presence of glutamine and amino acids had no beneficial effects on the development of 1-cell embryos to the blastocyst.

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Improvement of In Vitro Development of Bovine Embryos in a Medium Containing Selenium

  • Lee, J.H.;Park, J.H.;Choi, K.M.;Im, K.S.;Jin, D.I
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.2
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    • pp.170-173
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    • 2001
  • The objective of this study was to investigate the effect of selenium (Se) on in vitro development of bovine embryos. In CR1BSA, FBS-free medium, the bovine embryos could not proceed past the developmental block more efficiently to morula stage than in chemically undefined media. Addition of glutathione precursor, cysteine, with $\beta$-mercaptoethanol did not improve the development in chemically defined medium and neither did glutathione alone. Exogenous selenium improved the embryonic development to the morula and blastocyst stages at 6 days post-insemination (dpi) significantly (67.1% vs 57.5%, p<0.05), and blastocyst stage at 8 dpi (30.1% vs 20.5%, p>0.05). These improvements might be induced by elevated glutathione peroxidase activity due to addition of Se, and a possible mechanism of selenium to elevate the activity of glutathione peroxidase is discussed.

Effect of Defined KSOM Medium on the Development of 1-antitrypsin Transgenic Nuclear Transfer Bovine Embryos

  • M.M.U. Bhuiyan;J.K. Cho;G. Jang;Park, E.S.;S.K. Kang;Lee, B.C.;W.S. Hwang
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.74-74
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    • 2002
  • Production of u 1-antitrypsin ($\alpha$AT) in transgenic cows has a great value in the field of medicine. The present study was conducted to determine the effect of chemically defined KSOM media on in vitro development of bovine transgenic nuclear transfer (NT) embryos. An expression plasmid for human $\alpha$AT was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human $\alpha$AT target gene into a pcDNA3 plasmid. Cumulus cells as donor nuclei in NT were collected from a Holstein cow and transfected by lipid-mediated method using FuGene6 (Roche Molecular Biochemicals, USA) as reagent. GFP expressed cumulus cells were introduced into recipient oocytes under DIC microscopy equipped with FITC filter set. After electrical fusion and chemical activation, reconstructed embryos were cultured in 1) SOF + 0.8% BSA, 2) KSOM + 0.8% BSA, 3) KSOM + 10% FBS and 4) KSOM +0.01% PVA for 192 h at 39$^{\circ}C$ with 5% $CO_2$, 5% $O_2$ and 90% $N_2$in humidified condition. The development of the embryos was recorded and the GFP expression in blastocyst was determined under FITC filter. The average fusion rate was 73.8% (251/340; n=8). The development rates to 2-4 cells, morula, blastocysts and expression rates in blastocysts varied from 70.3 to 76.5%, 30.2 to 33.8%, 25.4 to 33.8% and 11.8 to 15.6%, respectively. The difference in development and expression rates of embryos among 4 culture groups was not significant (P>0.05). This study indicates that chemically defined KSOM medium is also able to support development of bovine transgenic NT embryos at similar rate of SOF or KSOM supplemented with BSA or serum.

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Standardization of Chemically Defined Medium for the Production of Mycelium and Basidiocarps in Flammulina velutipes (팽나무버섯 균사체 및 자실체 생산을 위한 화학합성배지의 최적화)

  • Song, Chi-Hyeun;Lee, Chang-Ho;Ahn, Jang-Hyuk;Hong, Bum-Shik;Yang, Han-Chul
    • The Korean Journal of Mycology
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    • v.23 no.1 s.72
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    • pp.53-60
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    • 1995
  • Nutritional requirements for the growth of Flammulina velutipes were studied. Mannitol, glutamic acid and ammonium nitrate were chosen for the maximum mycelial growth when various carbon and nitrogen sources tested. Optimum C : N ratio for the mycelial growth was 20 : 1. Potassium dihydrogen phosphate was selected among the phosphate sources. Magnesium sulphate and thiamine HCl stimulate mycelial growth. Final compositions of optimized chemically defined medium were 1.5% mannitol, 0.082% $NH_4NO_3$, 0.312% glutamic acid, 0.25% $KH_2PO_4$, 0.06% $MgSO_4{\cdot}7H_2O\;and\;0.3\;{\mu}g/l$ thiamin HCl. This medium not only support mycelial growth but also induce fruit body formation.

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Physiological and Nutritional Factors for Efficient Sporulation and Toxin Formation in Bacillus tthuringiensis

  • 이은희;반재구;김정일
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.522.2-522
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    • 1986
  • In order to optimally induce sporulation and toxin formation in Bacillus thuringiensis, exhaustion of specific nutrients as well as resuspension experiments were tried. Sporulation and toxin formation was most abunduntly occurred when the growth was limited by carbon source. It was also occurred in a resuspension medium containing only distilled water. Various environmental and physiological factors affecting the efficiencies of spore and toxin formation were examined in chemically defined media. As a result of these studies, a batch fermentation resulted in higher spore and toxin yield than ever reported

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Effect of Mammalian Spermatozoa on In Vitro Maturation of Porcine Germinal Vesicle Oocyte in Chemically Defined Medium

  • Kang, Sung-Ryoung;Kim, Byung-Ki
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.72-72
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    • 2002
  • Oocytes maturation, characterized by germinal vesicle (GV) breakdown, formation of the first meiotic spindle, expulsion of the first polar body and arrest in metaphase of second meiotic division (MII), occurs in preovulatory follicles in response to the surge of gonadotropin and leads to an ovulated oocyte in vivo. However, meiotic resumption in vitro occurs spontaneously following removal of cumulus-oocytes complexes (COCs) from the follicle. (omitted)

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Statistical Optimization of Growth Medium for the Production of the Entomopathogenic and Phytotoxic Cyclic Depsipeptide Beauvericin from Fusarium oxysporum KFCC 11363P

  • Lee, Hee-Seok;Song, Hyuk-Hwan;Ahn, Joong-Hoon;Shin, Cha-Gyun;Lee, Gung-Pyo;Lee, Chan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.138-144
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    • 2008
  • The production of the entomopathogenic and phytotoxic cyclic depsipeptide beauvericin (BEA) was studied in submerged cultures of Fusarium oxysporum KFCC 11363P isolated in Korea. The influences of various factors on mycelia growth and BEA production were examined in both complete and chemically defined culture media. The mycelia growth and BEA production were highest in Fusarium defined medium. The optimal carbon and nitrogen sources for maximizing BEA production were glucose and $NaNO_3$, respectively. The carbon/nitrogen ratio for maximal production of BEA was investigated using response surface methodology (RSM). Equations derived by differentiation of the RSM model revealed that the production of BEA was maximal when using 108 mM glucose and 25 mM $NaNO_3$.

Nutritional Conditions of Xylanase Production from Xylose Fermenting Yeast (Xylose 발효효모의 Xylanase 생성)

  • 배명애;김남순;방병호;서정훈
    • Microbiology and Biotechnology Letters
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    • v.17 no.2
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    • pp.85-87
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    • 1989
  • Cultural conditions for the formation of extracellular xylanase by Candida sp. X-6-41 were investigated. The xylanase was not produced in culture medium containing polypeptone or yeast extract as a nitrogen source, respectively, whereas the enzyme w8s produced in chemically defined medium containing (NH$_4$)$_2$SO$_4$as a sole nitrogen source. The xylanase production was affected by the amino acids such as isoleucine and tryptophan. The enzyme production of the strain was completely inhibited by the addition of isoleucine in the culture medium, but enhanced by tryptophan below the concentration of 25$\mu$g/$m\ell$.

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Examination of Metabolites Activating Production of Antibiotic in the Neomycin Producer, S. fradiae (Neomycin 생산균주 S. fradiae의 항생물질 생산을 활성화시키는 성분조사)

  • 김공환;구양모
    • KSBB Journal
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    • v.6 no.1
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    • pp.69-77
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    • 1991
  • When S. fradiae was cultured in S medium, it stavted to produce neomycin in the middle of stationary phase of growth. Antibitoic production is regulated not only by glucose but also by metabolites formed from glucose. A chemically defined minimal salt broth was developen for the study of metabolites activating produition of antibiotic in a neomycin producer. When growth and production or antibiotic in minimal salt broth was examined with a full grown or a vefctativc mycelium, the medium was found not to be good for the growth, but to be good enough for the production of antibiotic with a full grown mycelium. When many carbotlydrates, organic acids, or alcohol were supplmented with instead of glucose in the medium suspcndcn with a full grown mycelium, the amount of antibiotic produced in the medium containing fumaratc was 5 times more than that in the medium with glucose. Further study indicated that the medium is not good also for the growth but good for the production of antibiotic. The antibiotic produced in this medium was identified to be neomycin. The activation of the production of neomycin by fumarate was further confirmed in a complex medium. Fuinarate is suspected to initiate and to activate the biosynthesis of neomycin at the gene level.

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