• Title/Summary/Keyword: PHB

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Purification and Characterization of Poly(3-hydroxybutyrate) Depolymerase from a Fungal Isolate, Emericellopsis minima W2

  • Rhee, Young-Ha;Kim, Do-Young;Yun, Ji-Hye;Kim, Hyung-Woo;Bae, Kyung-Sook
    • Journal of Microbiology
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    • v.40 no.2
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    • pp.129-133
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    • 2002
  • The fungus, Emericellopsis minima W2, capable of degrading poly(3-hydroxybutyrate) (PHB) was isolated from a waste water sample. Production of the PHB depolymerase from E. minima W2 (PhaZ/ sub Emi/) was significantly repressed in the presence of glucose. PhaZ/ sub Emi/ was purified by column chromatography on Octyl-Sepharose CL-4B and Sephadex G-100. The molecular mass of the PhaZ/ sub Emi/), which consisted of a single polypeptide chain, was estimated to be 48.0 kDa by SDS-PAGE and its pI vague was 4.4. The maximum activity of the PhaZ/ sub Emi/ was observed at pH 9.0 and 55$\^{C}$. It was significantly inactivated by 1mM dithiothreitol, 2mM diisopropyl fluorphosphate, 0.1mM Tween 80, and 0.1 mM Triton X-l00, but insensitive to phenylmethylsulfonyl fluoride and N-ethylmaleimide. The PhaZ/ sub Emi/ efficiently hydrolyzed PHB and its copolyester with 30 mol% 3-hydroxyvalerate, but did not act on poly(3-hydroxyoctanoate). It also hydrolyzed p-nitrophenylacetate and p-nitrophenylbutyrate but hardly affected the longer-chain forms. The main hydrolysis product of PHB was identified as a dimer of 3-hydroxybutyrate.

High Cell Density Culture of Alcaligenes eutrophus and Poly-$\beta$-hydroxybutyrate Production by Optimization of Medium Compositions (배지조성 최적화를 통한 Alcaligenes eutrophus의 고농동 세포배양 및 Poly$\beta$-hydroxybutyrate 생산)

  • 이용우;유영제
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.401-406
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    • 1994
  • The medium compositions of Alcaligenes eutrophus were optimized for increasing PHB productivity. It is very important to optimize the concentrations of inorganic salts and trace eleme- nts as well as carbon and nitrogen sources to maximize cell growth rate and productivity. The fed-batch culture of Alcaligenes eutrophus by dual feeding of ammonia water and glucose under optimized initial medium concentrations was carried out. Glucose was fed manually according to glucose consumption rate and ammonia water by pH-stat. The final cell concentrations and PHB content in 30 hours were 122 g/l and 65% of dry cell weight(yielding 79 g of PHB/l), respectively and 2.64 g/l/hr of PHB production rate was obtained.

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Drug Release Characteristics of Biodegradable Polymers for Stent Coating (스텐트 코팅용 생분해성 고분자의 약물 방출 특성)

  • 강혜수;김진설;김동운;강병철;이봉희;김범수
    • KSBB Journal
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    • v.18 no.2
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    • pp.107-110
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    • 2003
  • Biodegradable polymers, poly(lactic-co-glycolic acid) (PLGA), poly(3-hydroxybutyrate) (PHB), and medium chain length polyhydroxyalkanoates (MCL-PHA) containing rose bengal (model drug) were coated onto the surface of stainless steel (stent materials) and their in vitro release characteristics were investigated. Drug release increased with; decreasing PLGA concentration, increasing rose bengal concentration, and Increasing dip-coating duration. The order of drug release from the polymer coating was: PHB > PLGA > MCL-PHA. These results suggest that drug release can be controlled by: changing the concentration and type of polymer, the drug concentration, and the dip-coating duration.

Studies on the Ternary Blends of Liquid Crystalline Polymer and Polyesters

  • Kim, Seong-Hun;Kang, Seong-Wook
    • Fibers and Polymers
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    • v.1 no.2
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    • pp.83-91
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    • 2000
  • Thermotropic liquid crystalline polymer made up of poly(p-hydroxybenzoate) (PHB)-poly(ethylene terephthalate)(PET) 8/2 copolyester, poly(ethylene 2,6-naphthalate) (PEN) and PET were mechanically blended to pursue the liquid crystalline phase of ternary blends. Complex viscosities of blends decreased with increasing temperature and PHB content. DSC thermal analysis indicated that glass transition temperature (Tg) and melting temperature (Tm) of blends increased with increasing PHB content. Both tensile strength and initial modulus increased with raising PHB content and take-up speed of monofilaments. In the WAXS diagram, only PEN crystal reflection at 2Θ=$15.5^{\circ}C$ appeared but PET crystal reflection was not shown in all compositions. The degree of transesterification and randomness of blends increased with blending time but sequential length of both PEN and PET segment decreased.

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Intravenous levetiracetam versus phenobarbital in children with status epilepticus or acute repetitive seizures

  • Lee, Yun-Jeong;Yum, Mi-Sun;Kim, Eun-Hee;Ko, Tae-Sung
    • Clinical and Experimental Pediatrics
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    • v.59 no.1
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    • pp.35-39
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    • 2016
  • Purpose: This study compared the efficacy and tolerability of intravenous (i.v.) phenobarbital (PHB) and i.v. levetiracetam (LEV) in children with status epilepticus (SE) or acute repetitive seizure (ARS). Methods: The medical records of children (age range, 1 month to 15 years) treated with i.v. PHB or LEV for SE or ARS at our single tertiary center were retrospectively reviewed. Seizure termination was defined as seizure cessation within 30 minutes of infusion completion and no recurrence within 24 hours. Information on the demographic variables, electroencephalography and magnetic resonance imaging findings, previous antiepileptic medications, and adverse events after drug infusion was obtained. Results: The records of 88 patients with SE or ARS (median age, 18 months; 50 treated with PHB and 38 with LEV) were reviewed. The median initial dose of i.v. PHB was 20 mg/kg (range, 10-20 mg/kg) and that of i.v. LEV was 30 mg/kg (range, 20-30 mg/kg). Seizure termination occurred in 57.9% of patients treated with i.v. LEV (22 of 38) and 74.0% treated with i.v. PHB (37 of 50) (P=0.111). The factor associated with seizure termination was the type of event (SE vs. ARS) in each group. Adverse effects were reported in 13.2% of patients treated with i.v. LEV (5 of 38; n=4, aggressive behavior and n=1, vomiting), and 28.0% of patients treated with i.v. PHB (14 of 50). Conclusion: Intravenous LEV was efficacious and safe in children with ARS or SE. Further evaluation is needed to determine the most effective and best-tolerated loading dose of i.v. LEV.

Expression pattern of prohibitin, capping actin protein of muscle Z-line beta subunit and tektin-2 gene in Murrah buffalo sperm and its relationship with sperm motility

  • Xiong, Zhaocheng;Zhang, Haihang;Huang, Ben;Liu, Qingyou;Wang, Yingqun;Shi, Deshun;Li, Xiangping
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.11
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    • pp.1729-1737
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    • 2018
  • Objective: The aim of the current study is to investigate the relationship between prohibitin (PHB), capping actin protein of muscle Z-line beta subunit (CAPZB), and tektin-2 (TEKT2) and sperm motility in Murrah buffalo. Methods: We collected the high-motility and low-motility semen samples, testis, ovary, muscle, kidney, liver, brain and pituitary from Murrah buffalo, and analysed the expression of PHB, CAPZB, and TEKT2 in mRNA (message RNA) and protein level. Results: Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) result showed that the expression of PHB was higher and CAPZB, TEKT2 were specifically expressed in testis as compared to the other 6 tissues, and that in testis, the expression of TEKT2 was higher than that of CAPZB and PHB. Immunohistochemistry test revealed that all three genes were located on the convoluted seminiferous tubule and enriched in spermatogenic cells. Both qRT-PCR and Western Blot results showed that the expression levels of PHB, CAPZB, and TEKT2 were significantly lower in the low-motility semen group compared to the high-motility semen group (p<0.05). Conclusion: The expression levels of PHB, CAPZB, and TEKT2 in Murrah buffalo sperm have a high positive correlation with sperm motility. And the three genes may be potential molecular markers for the decline of buffalo sperm motility.

Effect of Cosubstrate on tile Production of Poly(3-Hydroxybutyric-Co-3-Hydroxyvaleric) Acid from Glucose by Pseudomonas sp, HJ (Pseudomonas sp. HJ에 의한 포도당으로부터 Poly(3-Hydroxybutyric-Co-3-Hydroxyvaleric) Acid의 생합성에 대한 보조기질의 영향)

  • 손홍주;고명선이상준
    • KSBB Journal
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    • v.11 no.5
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    • pp.586-592
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    • 1996
  • Poly(3-hydroxybutyric-co-3-hydroxyvaleric) acid(PHB/HV) copolymer synthesis by Pseudomonas sp. HJ from glucose and cosubstrate was investigated. Taxonomic analysis suggested that Pseudomonas sp. HJ was best marched to Pseudomonas picketti having 78.8% similarity. Pseudomonas sp. HJ produced PHB/HV copolymer containing 60.8 mol% HV and 44.9 mol% HV when supplied with hexadecane and propionic acid as a cosubstrate, respectively. The HV composition in PHB/HV copolymer was controlled by varying the concentration of hexadecane and propionic acid. Propionic acid added after 24 hours of incubation was incorporated as the HV monomer in the PHB/HV copolymer up to 49.6 mol%.

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Effect of Oxygen Composition on Polyhydroxybutyrate Synthesis by Alcaligenes eutrophus at Various Pressures (Alcaligenes eutrophus에 의한 Polyhydroxybutyrate의 합성에 관한 산소효과)

  • Kim, Kyeo-Keun;Shin, Sun Kyoung;Kwon, Hyo-Shik
    • Analytical Science and Technology
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    • v.10 no.1
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    • pp.75-81
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    • 1997
  • Poly-${\beta}$-hydroxybutyrates(PHB) are well known intracellular biopolymer which are completely degraded to carbon dioxide and water in the landfill. The pressurized fermentation method was used to increase the cell growth of Alcaligenes eutrophus and productivity of PHB. The experimental data were analyzed in terms of carbon source and gas composition effects at various elevated pressures and temperatures. The results reveal that the flow rate of hydrogen gas of 0.0075vvm for the culture is better PHB production than the no hydrogen flow when fructose was used as a sole carbon source. The higher yields and productivities of PHB biosynthesized by A. eutrophus were obtained when the oxygen composition was changed from 2% to 8% at 6atm and $30^{\circ}C$.

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Enhanced PHB Accumulation in Photosystem- and Respiration-defective Mutants of a Cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis sp. PCC 6803의 에너지 대사 결함 돌연변이 균주에서의 Poly(3-hydroxybutyrate) 축적량 증진)

  • Kim Soo-Youn;Choi Gang Guk;Park Youn Il;Park Young Mok;Yang Young Ki;Rhee Young Ha
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.67-73
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    • 2005
  • Photoautotrophic bacteria are promising candidates for the production of poly(3-hydroxybutyrate) (PHB) since they can address the critical problem of substrate costs. In this study, we isolated 25 Tn5-inserted mutants of the Synechocystis sp. PCC 6803 which showed enhanced PHB accumulation compared to the wild-type strain. After 5-days cultivation under nitrogen-limited mixotrophic conditions, the intracellular levels of PHB content in these mutants reached up to $10-30\%$ of dry cell weight (DCW) comparable to $4\%$ of DCW in the wild-type strain. Using the method of inverse PCR, the affected genes of the mutants were mapped on the completely known genome sequence of Synechocystis sp. PCC 6803. As a result, the increased PHB accumulation in 5 mutants were found to be resulted from defects of genes coding for NADH-ubiquinone oxidoreductase, O-succinylbenzoic-CoA ligase, photosystem II PsbT protein or histidine kinase, which are involved in photosystem in thylakoid inner membrane of the cell. The values of $NAD(P)H/NAD(P)^+$ ratio in the cells of these mutants were much higher than that of the wild-type strain as measured by using pulse-amplitude modulated fluorometer, suggesting that PHB synthesis could be enhanced by increasing the level of cellular NAD(P)H which is a limiting substrate for NADPH-dependent acetoacetyl-CoA reductase. From these results, it is likely that NAD(P)H would be a limiting factor for PHB synthesis in Synechocystis sp. PCC 6803.