• Title/Summary/Keyword: Beta-hydroxybutyrate

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Isolation and Characterization of a Methylotroph Producing 3-hydroxybutyrate-3-hydroxyvalerate Copolymer

  • JUNG HOE KIM;KIM, PIL;SEON WON KIM;GYUN MIN LEE;HYUN SOO LEE
    • Journal of Microbiology and Biotechnology
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    • v.5 no.3
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    • pp.167-171
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    • 1995
  • A bacterial strain C-02 using methanol as a carbon source was isolated from Gumi Industrial Estate and selected based on its rapid growth and capability of poly-$\beta$-hydroxybutyrate accumulation. Characteristics of strain C-02 showed that it belongs to the Methylococcaceae family, Type II subgroup. Strain C-02 could incorporate valerate into the PHB chain to form 3-hydroxybutyrate and 3-hydroxyvalerate (P(3HB-co-3HV)). Among various nutrient limitation tests, the nitrogen limitation test resulted in the highest content of P(3HB-co-3HV) per dry cell weight, 50$%$. Under the nitrogen limited condition, the average molecular weight of P(3HB-co-3HV) obtained was determined to be approximately $2.8\times 10^5$ daltons.

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주정증류 폐액을 이용한 Actinobacillus sp. EL-9로부터 Poly-$\beta$-Hydroxybutyrate의 생산 및 폐약의 처리

  • 손홍주;이상준
    • Microbiology and Biotechnology Letters
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    • v.24 no.3
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    • pp.352-356
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    • 1996
  • Alcoholic distillery wastes are utilized as dual purposes to produce PHB in lower production cost and to reduce the amount of waste to be treated. In this study, various attempts were made to increase PHB production under various conditions by Actinobacillus sp. EL-9 in a shaker culture. The addition of glucose, NH$_{4}$NO$_{3}$ to alcoholic distillery wastes slightly promoted cell mass and PHB production. Enzyme hydrolysis of alcoholic distillery wastes increased the production of PHB than that of untreated waste and acid hydrolysis treatment. The PHB weight in alcoholic distillery wastes was 1.91 g/l. Fermentation process of PHB production reduced the amount of COD value up to 54%, which reduced organic loading rate and capacity of activated sludge system.

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Quorum Sensing of Rhodobacter sphaeroides Negatively Regulates Cellular Poly-$\beta$-Hydroxybutyrate Content Under Aerobic Growth Conditions

  • Lee, Jeong-K.;Kho, Dhong-Hyo;Jang, Ji-Hee;Kim, Hye-Sun;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.477-481
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    • 2003
  • The community escape response of Rhodobacter sphaeroides is exerted through the action of CerR and CerI, which code for a LuxR-type regulatory protein and acylhomoserine lactone synthase, respectively. Deletion of chromosomal DNA including cerR and cerI (mutant RI) or insertional interruption of cert (mutant AP3) resulted in two-fold increase in the cellular poly-${\beta}$-hydroxybutyrate (PHB) content In comparison with the wild-type under aerobic growth conditions. The PHB synthase (PhbC) activities of the cer mutants were doubled, and the enzyme expression was regulated at the level of phbC transcription. Thus, CerR, possibly in response to autoinducer (AI), appears to modulate the PHB content of aerobically grown cells by downregulating phbC transcription.

Simultaneous Determination of Plasma Lactate, Pyruvate, and Ketone Bodies following tert-Butyldimethylsilyl Derivatization using GC-MS-SIM

  • Yoon, Hye-Ran
    • Biomedical Science Letters
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    • v.21 no.4
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    • pp.241-247
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    • 2015
  • Lactate and ketone bodies are considered biological markers for ketosis and several inherited metabolic disorders. In the current study, the specific ratios of lactate and ketone bodies as analytical tools for differential diagnosis of various lactic acidosis were devised. The study included a protein precipitation step following tert-butyldimethylsilyl derivatisation. Total run time was approximately 30 min including sample preparation and GS/MS analysis. The limits of detection were below 0.1 pg/mL over the targeted 4 analytes. The calibration curve was linear over the concentration range of $0.001{\sim}250{\mu}g/mL$ for pyruvate, beta-hydroxybutyrate, and acetoacetate ($R^2$ > 0.99). Inter-day accuracy and precision were 87.7~94.8% with RSD of 2.5~5.7% at 2 levels. Absolute recoveries (%) of target analytes were 87.0~98.4%. The method was validated for the quantification of lactate and ketone bodies for differentiation of lactic acidosis.

Metabolic Flux Analysis of a Poly-${\beta}$-hydroxybutyrate Producing Cyanobacterium, Synechococcus sp. MA19, Grown under Photoautotrophic Conditions

  • Nishioka, Motomu;Nishiuma, Hajime;Miyake, Masato;Asada, Yasuo;Shimizu, Kazuyuki;Taya, Masahito
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.5
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    • pp.295-302
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    • 2002
  • To understand the utilization property of light energy, Synechococcus sp. MA19, a poly-${\beta}$-hydroxybutyrate (PHB) producer, was cultivated at the different incident light intensities of 15.3, 50.0 and 78.2 W/$m^2$ using media with and without phosphate. From the results of metabolic flux analysis, it was found that the cell yield based on ATP synthesis was estimated as $3.5{\times}10^{-3}$ kg-biomass/mol-ATP in these cultures. Under the examined conditions, there were no significant differences in the efficiency of light energy conversion to chemical energies estimated as ATP synthesis and reducing potential (NADH + NADPH) formation whether the PHB synthesis took place or not. The energy converted from light to ATP was kept relatively high around the energy absorbed by the cells of $2.5-3.0{\times}10^{6} J\;h^{-1}\;kg^{-1}$, whereas the energy of reducing potential was hardly changed in the examined range of the energy absorbed by the cells.

Maximization of Poly-$\beta$-Hydroxybutyrate Accumulation by Potassium Limitation in Methylobacterium organophilum and Its Related Metabolic Analysis

  • Kim, Seon-Won;Kim, Pil;Kim, Jung-Hoe
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.140-146
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    • 1999
  • When methanol was the sole carbon source, Methylobacterium organophilum NCIB 11278, a facultative methylotroph, accumulated Poly-$\beta$-hydroxybutyrate (PHB) as 59% (w/w) of dry cell weight under potassium limitation, 37% under sulfate limitation, and 33% under nitrogen limitation. Based on a stoichiometric analysis of PHB synthesis from methanol, it was suspected that PHB synthesis is accompanied by the overproduction of energy, either 6-10 ATP and 1 $FADH_2$ or 6 ATP and 3 NADPH to balance the NADH requirement, per PHB monomer. This was confirmed by observation of increased intracellular ATP levels during PHB accumulation. The intracellular ATP with limited potassium, sulfate, and ammonium increased to 0.185, 0.452, and 0.390 $\mu$moles ATP/g Xr (residual cell mass) during PHB accumulation, respectively. The intracellular ATP level under potassium limitation was similar to that when there was no nutrient limitation and no PHB accumulation, 0.152- 0.186 $\mu$moles ATP/g Xr. We propose that the maximum PHB accumulation observed when potassium was limited is a result of the energy balance during PHB accumulation. Microorganisms have high energy requirements under potassium limitation. Enhanced PHB accumulation, in ammonium and sulfate limited conditions with the addition of 2,4-dinitrophenol, which dissipates surplus energy, proves this assumption. With the addition of 1 mM of 2,4-dinitrophenol, the PHB content increased from 32.4% to 58.5% of dry cell weight when nitrogen limited and from 15.1 % to 31.0% of dry cell weight when sulfate limited.

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Effect of the Addition of β-Hydroxybutyrate to Chemically Defined Maturation Medium on the Nuclear Maturation, Sperm Penetration and Embryonic Development of Porcine Oocytes In vitro

  • Endo, R.;Ishii, A.;Nakanishi, A.;Nabenishi, H.;Ashizawa, K.;Tsuzuki, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.11
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    • pp.1421-1426
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    • 2010
  • We investigated the effects of various concentrations of ${\beta}$-hydroxybutyrate (BHB, 0, 0.1, 1 and 10 mM), a ketone body, added to chemically-defined maturation medium with or without energy substrates (glucose, pyruvate and lactate) on nuclear maturation rates up to the metaphase stage of the second meiotic division (M-II stage). In addition, we also assessed the influence of BHB on glutathione content, sperm penetration rate and embryonic development up to the blastocyst stage of oocytes matured under the presence of these energy substrates. Nuclear maturation rates up to the M-II stage of oocytes matured with BHB in each concentration group did not show a significant increase compared with the control (0 mM) groups in both the presence and absence of energy substrates. Although glutathione contents were not significantly different in each BHB concentration group, the sperm penetration rate in the 1 mM BHB group was significantly higher (p<0.05) and the embryonic development rate of oocytes up to the blastocyst stage was significantly lower (p<0.05) than the respective values of the control groups. These results suggest that BHB added to a chemically-defined maturation medium may stimulate sperm penetration while inhibiting embryonic development of porcine oocytes.

Optimization of Fed-Batch Fermentation for Production of Poly-$\beta$-Hydroxybutyrate in Alcaligenes eutrophus

  • Lee, In-Young;Choi, Eun-Soo;Kim, Guk-Jin;Nam, Soo-Wan;Shin, Yong-Cheol;Chang, Ho-Nam;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.146-150
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    • 1994
  • Production of poly-$\beta$-hydroxybutyrate (PHB) in fed-batch fermentation was studied. Utilization of carbon for PHB biosynthesis was investigated by using feeding solutions with different ratios of carbon to nitrogen (C/N). It was observed that at a high C/N ratio carbon source was more preferably utilized for PHB accumulation while its consumption for cellular metabolism appeared to be more favored at a low C/N value. A high cell concentration (184 g/l) was achieved when ammonium hydroxide solution was fed to control the pH, which was also utilized as the sole nitrogen source. For the mass production of PHB, two-stage fed-batch operations were carried out where PHB accumulation was observed to be stimulated by switching the ammonium feeding mode to the nitrogen limiting condition. A large amount of PHB (108 g/l) was obtained with cellular content of 80% within 50 hrs of operation.

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Effects of Nitrogen and Oxygen Supply on Production of $Poly-{\beta}-Hydroxybutyrate$ in Azotobacter chroococcum

  • Lee, In-Young;Stegantseva, Ellen-M.;Savenkova, Ludmila;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.5 no.2
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    • pp.100-104
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    • 1995
  • Production of $poly-{\beta}-hydroxybutyrate$ (PHB) in a strain of Azotobacter chroococcum, a nitrogen-fixing bacteria, was investigated at various levels of nitrogen and oxygen. Feeding nitrogen source increased both cell growth and PHB accumulation. Oxygen supply appeared to be one of the most important operating parameters for PHB production. Both cell growth and PHB accumulation increased with the sufficient supply of air in the fed-batch fermentation of the strain. However, it was also noted that keeping the oxygen level under limited condition was critical to achieve high PHB productivity. A high titer of PHB (52 g/l) with a high cellular content (60%) was obtained after 48 hr of fed-batch operation by controlling the oxygen supply. Dual limitation of nitrogen and oxygen did not further increase the PHB accumulation probably due to the greater demand for reducing power and ATP for nitrogen fixation.

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