• Title/Summary/Keyword: Inositol

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Purification of Inositol Triphosphate Kinase from Bovine Brain (소의 뇌로부터 Inositol Triphosphate Kinase의 정제)

  • Kim, Jung-Hye;Lee, Jae-Tae
    • Journal of Yeungnam Medical Science
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    • v.13 no.1
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    • pp.46-58
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    • 1996
  • Inositol 1,4,5-triphosphate($InsP_3$) is a second messenger for mobilizing intracellular $Ca^{2+}$. It can be dephosphorylated by soluble and particulate forms on $InsP_3$ 5-phosphatase, or phosphorylated to produce inositol 1,3,4,5-tetrakisphosphate($InsP_3$) by $InsP_3$ 3-kinase. These enzymes represent possible targets for the regulation of the $InsP_3/InsP_4$ signal. $InsP_3$ 3-kinase which catalyses th ATP-dependent phosphorylation of $InsP_3$ was purified from bovine brain tissue. All operation were carried out at $4^{\circ}C$. Fresh tissure was homogenized and centrifuged. The supernatant was pooled. Proteins were precipitated from 10% polyethylene glycol, and suspended solution was applied to DEAE cellulose column for chromatography. As the result of above procedure, two isozymes of $InsP_3$ 3-kinase, I and II were obtained. Each isozyme was applied to Matriz green gel, Calmodulin-Affigel 15 column and subsequent phenyl-TSK HPLC column. Specific activites(SA) and fold of puriety were observed at each purification step of chromatography. At DEAE cellulose chromatography, SA were I, 0.6 and II, 4.8 nM/min/mg, and folds were I, 17.2 and II, 16.6. At Matrix green gel chromatography, SA were I, 18 and II, 11 nM/min/mg, folds were I, 62.1 and II, 38.0. At calmodulin-Affigel 15 column chromatography, SA were I, 19 and II, 13 nM/min/mg, folds were I, 65.5 and II, 44.8. Finally $InsP_3$ kinase I and II were purified 3,103-fold and 2,310-fold, and SA were I, 900 and II, 670 nM/min/mg, respectively. SDS-polyacrylamide gel electrophoresis elucidated 3 distinct fractions of Mr of 145,000, 85,000 and 69,500 from isozyme I, and 2 distinct fractions of Mr of 79,000 and 57,000 from isozyme II.

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The characteristics of phosphorus in major Korean soils -II. The characteristics of organic phosphorus (우리나라 주요토양중(主要土壤中) 인산(燐酸)의 특성(特性)에 관한 연구 -II. 유기인산(有機燐酸)의 분획정량치(分劃定量値)를 중심(中心)으로)

  • Hong, Jung-Kook;Hong, Chong-Woon
    • Korean Journal of Soil Science and Fertilizer
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    • v.12 no.1
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    • pp.35-41
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    • 1979
  • A study was conducted to investigate the status of organic phosphorus in soils originated from different parent materials and under different uses(Virgin, rice soil, upland). The results are summarized as a following. 1. Regardless of parent materials and land uses, the contents of different forms of organic phosphorus are in the order of unidentified organic-P>Inositol-P>Sugar-P>Lipid-P. 2. In case of Inositol-p, the contents were in the order of rice soil>upland>virgin soil. 3. The contents of Sugar-P and Lipid-P in cultivated soils are higher than in uncultivated soil. 4. RNA and DNA are not identified in any soil when detected by ultra-violet absorption method. 5. There is highly significant correlation between the content of soil organic matter and that of organic phosphate. However, there is remarkable difference in the pattern of correlation between cultivated soils and uncultivated soils. 6. Comments were made on the factors on plant availability of various organic phosphorus.

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Myo-inositol increases the plating efficiency of protoplast derived from cotyledon of cabbage (Brassica oleracea var. capitata)

  • Jie, Eun-Yee;Kim, Suk-Weon;Jang, Hye-Rim;In, Dong-Su;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.69-76
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    • 2011
  • This study describes the effect of myo-inositol on sustained cell division and plant regeneration from cotyledon-derived protoplast of cabbage (Brassica oleracea var. capitata). Freshly isolated protoplasts were cultured in modified Murashige and Skoog (MS) medium removed ammonia ions and containing $0.4\;mg\;l^{-1}$ thiamine HCl, $100\;mg\;l^{-1}$ myo-inositol, $2\;mgl^{-1}$ 2,4-D, $0.5\;mgl^{-1}$ BA, $30\;gl^{-1}$ sucrose and several concentrations of myo-inositol (2, 4, 6, 8, 10% (w/v)) as an osmotic stabilizer. After 3 weeks of culture in the dark at $25^{\circ}C$, the plating efficiency of cabbage protoplasts reached to $22.5{\pm}2.9%$ when cultured in modified MS medium supplemented with $2\;mgl^{-1}$ 2,4-D, $0.5\;mgl^{-1}$ BA, $30\;gl^{-1}$ sucrose and 8% (w/v) of myo-inositol at a density of $2{\times}10^5$ protoplasts/ml. Rapidly growing cell colonies after 3 weeks of culture were transferred to the same culture medium removed osmoticum. To induce shoot regeneration from calluses, calluses with about 2 mm in diameter were transferred to the MS medium containing $2\;mgl^{-1}$ BA and $0.5\;mgl^{-1}$ NAA. After further three weeks of incubation onto the medium in the light, green shoots were formed on the surface of calluses at a frequency of 30%. Upon transfer to half-strength MS basal medium, roots were formed onto the bottom of regenerated shoots without auxin treatments. These regenerated plantlets were successfully acclimatized to soil transfer, grown to normal mature plants. The cabbage protoplast culture system established in this study could be applied for production of somatic hybrids or cybrids by asymmetric protoplast fusion and mass proliferation of elite somatic clones of cabbage.

Studies on the Inhibitory Substance of Yeast Growth. Part III. Effect of the Vitamins on the Yeaststatic Activity of Astradix-P. (항효모성물질에 관한 연구 (제삼보) Vitamin이 Astradix-P의 작용에 미치는 영향)

  • 서정훈;이인구;송방호
    • Microbiology and Biotechnology Letters
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    • v.1 no.2
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    • pp.89-92
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    • 1973
  • In the previous paper the biological characteristics and some reaction mechanisms of Astradix-P, yeaststatic substance, were reported. Especially yeaststatic activity of Astradix-P was anta-gonistically inhibited by alkaline amino acids, arginine, lysine and histidine, which were added as a nitrogen source in the yeast growing medium. In this paper the effects of alkaline nitrogen containing substance and several vitamins on the yeaststatic activity were investigated. The antagonistic action of alkaline nitrogen containing substance; adenine and vitamins; thiamine, riboflavin, pyridoxin, cobalamin, nicotinic acid, folic acid, biotin, p-aminobenzoic acid, inositol, and pantothenic acid to Astradix-P were not observed, thus evidencing that the yeaststatic activity of Astradix-P was not inhibited by a alkaline nitrogen containing substance and several vitamins.

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Screening of Phytase Overproducing Strains in Aspergillus spp. by UV Mutagenesis

  • Lee, Eung-Suek;Paik, In-Kee;Hahm, Young-Tae
    • Mycobiology
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    • v.28 no.3
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    • pp.119-122
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    • 2000
  • Phytases (myo-inositol hexakisphosphate phosphohydrolase; EC 3.1.3.8) are enzymes which catalyze the hydrolisys of phytate into myo-inositol and inorganic phosphates. Phytases are found in plants and a variety of microorganisms. Aspergillus species were treated with 254 nm of UV irradiation for the screening of phytase overproducing mutant strains. At 15 minute irradiation, the survivals of population were less than 5%, and UV irradiation time was decided at 20 minute for the isolation of mutant strains. Four UV mutant strains in A. oryzae (YUV-47, -169, -341, -511) and six in A. ficuum (FUV-17, -36, -69, -193, -317, -419) were isolated on PSM media containing ammonium phosphate. The specific enzyme activities of A. ficuum mutants are 110 to 140% higher than that of wild type.

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Purification of Phytase from Aspergillus ficuum and Production of Anti-phytase Antibody (Aspergillus ficuum의 Phytase의 정제와 Anti-phytase 항체생산)

  • Kim, Keun
    • The Korean Journal of Mycology
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    • v.27 no.4 s.91
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    • pp.299-303
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    • 1999
  • Phytase(myo-inositol-hexakis phosphate 3-phosphohydrolase, E C 3.1.3.8) sequentially hydrolyzes phytate to myo-inositol and inorganic phosphate. Phytase of Aspergillus ficuum was purified to homogeneity using ultrafiltration, cation exchange column and anion exchange column. It's molecular weight is estimated as around 90,000 by SDS-PAGE. Antibody against the phytase was produced by immunizing mice with the purified phytase. The titer of the antibody was determined to be 1/25,000.

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Protoplast Formation and Regeneration of Ganoderma lucidum (Ganoderma lucidum의 원형질체 형성과 재생)

  • 박영도;박경숙;이재성
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.311-314
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    • 1985
  • Ganodema lucidum protoplasts were formed by the treatment of Novozym 234. The osmotic stabilizers such as mannitol were effective enough to produce protoplasts up to 10$^{6}$ $m\ell$. For regeneration, however, MgSO$_4$.7$H_2O$ was suitable. When inositol and sucrose were employed as osmotic stablizers, the regeneration ratio reached to 0.26%. Overlay of Streptomycin sulfate added agar was required to prevent bacterial contamination.

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Effects of Protein Kinases on Phospholipase C Activation and Intracellular $Ca^{2+}$ Mobilization Induced by Endothelin-1 (Endothelin-1에 의한 phospholipase C 활성화와 세포내 $Ca^{2+}$ 이동에 미치는 protein kinase들의 효과)

  • 조중형;김현준;이윤혜;박진형;장용운;이승준;이준한;윤정이;김창종
    • YAKHAK HOEJI
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    • v.44 no.2
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    • pp.162-168
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    • 2000
  • To investigate the effects of protein kinases on endothelin-1-induced phospholipase C activation and $Ca^{2+}$ mobilization in Rat-2 fibroblast, we measured the formation of inositol phosphates and intracellular $Ca^{2+}$ concentration with [$^3$H]inositol and Fura-2/AM, respectively. Endothelin-1 dose-dependently activated phospholipase C and increased intracellular $Ca^{2+}$ concentration. Protein kinase C activator PMA, significantly inhibited both phospholipase C activity and $Ca^{2+}$ mobilization induced by endothelin-1. Tyrosine kinase inhibitor, genistein, inhibited both. On the other hand, cyclic nucleotide (cAMP and cGMP) did not have any influence on the signaling pathway of phospholipase C-Ca$^{2+}$ mobilization induced by endothelin-1. These results suggest that protein kinase C and tyrosine kinase counteract on the signaling pathway of phospholipase C-Ca$^{2+}$ mobilization induced by endothelin-1 in Rat-2 fibroblast. fibroblast.

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Role of Type 1 Inositol 1,4,5-triphosphate Receptors in Mammalian Oocytes

  • Yoon, Sook Young
    • Development and Reproduction
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    • v.23 no.1
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    • pp.1-9
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    • 2019
  • The ability of oocytes to undergo normal fertilization and embryo development is acquired during oocyte maturation which is transition from the germinal vesicle stage (GV), germinal vesicle breakdown (GVBD) to metaphase of meiosis II (MII). Part of this process includes redistribution of inositol 1, 4, 5-triphosphate receptor (IP3R), a predominant $Ca^{2+}$ channel on the endoplasmic reticulum membrane. Type 1 IP3R (IP3R1) is expressed in mouse oocytes dominantly. At GV stage, IP3R1 are arranged as a network throughout the cytoplasm with minute accumulation around the nucleus. At MII stage, IP3R1 diffuses to the entire cytoplasm in a more reticular manner, and obvious clusters of IP3R1 are observed at the cortex of the egg. This structural reorganization provides acquisition of $[Ca^{2+}]_i$ oscillatory activity during fertilization. In this review, general properties of IP3R1 in somatic cells and mammalian oocyte are introduced.