• Title/Summary/Keyword: PHB

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Cultivation of Alcaligenes eutrophus Transforming Cloned phbC Gene from Alcaligenes latus for Production of P(3-hydroxybutyrate-4-hydroxybutyrate) Containing High Molar Fraction of 4-Hydroxybutyrate (phbC 유전자가 도입된 형질전환 Alcaligenes eutrophus를 이용한 고분율 4-hydroxybutyrate 함유 P(3-hydroxybutyrate-4-hydroxybutyrate)의 생산)

  • Gang, Myeong-Sin;Jeong, Yeong-Mi;Lee, Yong-Hyeon
    • KSBB Journal
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    • v.14 no.4
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    • pp.422-428
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    • 1999
  • A transformat Alcaligence eutrophus GA5 harboring phbC gene from A. latus was cultivated for production of Poly(3-hydroxybutyrate-4-hydroxybutyrate)[P(3HB-4HB)] containing high molar fraction of 4-hydroxybutyrate(4HB)] containing high molar fraction of 4-hydroxybutyrate(4HB). Transformation did not influenced significantly on total cell growth, on total cell growth, concentration, and content of P(3HB-4HB), however, significantly influenced on 4HB molar fraction in P(3HB-4HB) increasing from 12.3 to 23.5 mol% after 48 h cultivation in two-stage using 1.0%(W/V) of ${\gamma}$-butyrolactone as a precursor compare to parent strain. Above increment may be due to the accelerated polymerization between 3HB and 4HB converted from precusor compound by amplified phbC gene. Citrate increased remarkbly total cell mass and P(3HB-4HB) concentration, but did not influenced on the molar fraction of 4HB, meanwhile, magnesium ion influenced on P(3HB-4HB) concentration and 4HB molar fraction significantly. The two-stage cultivation method was modified, in such a way minimizing P(3HB) accumulated inside of cell grown at first-stage, consquently, 26.3% of P(3HB-4HB) containing 61.0 mol% of 4HB fraction was obtained after 72hr. Furthermore, semi-homopolymeric P(4HB) containing 92.0 mol% of 4Hb was obtained, and its structure was confirmed by $^1$H-NMR.

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An Environmentally Friendly and Efficient Method for Extraction of PHB Biopolymer with Non-Halogenated Solvents

  • Aramvash, Asieh;Gholami-Banadkuki, Narges;Moazzeni-Zavareh, Fatemeh;Hajizadeh-Turchi, Samira
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1936-1943
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    • 2015
  • The present study developed an efficient and environmentally friendly method for recovering polyhydroxybutyrate (PHB) from Cupriavidus necator. Several non-halogenated solvents were tested and it was found that butyl acetate and ethyl acetate are powerful solvents for the biopolymer. Testing was performed to examine the effects of temperature (25℃ until temperature below solvent boiling points) and heating incubation time (0-60 min) on the two solvents. Butyl acetate had a higher recovery level (96%) and product purity (up to 99%) than ethyl acetate at 103℃ and a heating incubation time of 30 min. Under these conditions, PHB recorded the highest molecular weight of 1.4 × 106 compared with the standard procedure (i.e., recovery using chloroform). The proposed strategy showed that butyl acetate is a good alternative to halogenated solvents such as chloroform for recovery of PHB.

Metabolic Analysis of Poly(3-Hydroxybutyrate) Production by Recombinant Escherichia coli

  • WONG, HENG HO;RICHARD J. VAN WEGEN;JONG-IL CHOI;SANG YUP LEE
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.593-603
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    • 1999
  • Poly(3-hydroxybutyrate) (PHB) production by fermentation was examined under both restricted- and ample-oxygen supply conditions in a single fed-batch fermentation. Recombinant Escherichia coli transformed with the PHB production plasmid pSYLl07 was grown to reach high cell density (227 g/l dry cell weight) with a high PHB content (78% of dry cell weight), using a glucose-based minimal medium. A simple flux model containing 12 fluxes was developed and applied to the fermentation data. A superior closure (95%) of the carbon mass balance was achieved. When the data were put into use, the results demonstrated a surprisingly large excretion of formate and lactate. Even though periods of severe oxygen limitation coincided with rapid acetate and lactate excretion, PHB productivity and carbon utilization efficiency were not significantly impaired. These results are very positive in reducing oxygen demand in an industrial PHA fermentation without sacrificing its PHA productivity, thereby reducing overall production costs.

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Antimicrobial polyhydroxybutyrate submicron fiber mat loaded with extract of Hypericum perforatum

  • Beran, Milos;Horna, Ales;Vorisek, Viktor;Berkova, Eliska;Korinkova, Radka;Trousil, Vojtech;Hrubanova, Marketa
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.257-270
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    • 2022
  • The aim of this work was to prepare a new biodegradable polyhydroxybutyrate (PHB) submicron fiber mat loaded with hypericin-rich Hypericum perforatum raw extract by centrifugal spinning technology, an alternative approach to the traditional method of electrospinning to fabricate nanofibers or microfibers from solutions at high speed and low cost. Hypericins in methanol/acetone extract of H. perforatum were determined by UHPLC-MS/MS and HPLC/PDA. Submicron fiber mats composed of pure PHB or PHB enriched with H. perforatum extract were prepared using a pilot plant demonstrator for the centrifugal spinning technology and characterized by SEM. Singlet oxygen production was quantified by the 1,3-diphenylisobenzofuran (DPIBF) method in hexane. The results proved a significant production of singlet oxygen by the prepared submicron fiber mat. We also found a significant antibacterial activity against the bacterial strain Escherichia coli CCM 5417 by a method in accordance with JIS Z 2801/ISO 22196 standards. The H. perforatum extract-enriched PHB submicron fiber mats showed potential for the development of self-cleaning and antimicrobial air filters.

The Relationship between Prohibitin 1 Expression, Hepatotoxicity Induced by Acetaminophen, and Hepatoprotection by S-Adenosylmethionine in AML12 Cells

  • Eunhye Cho;Soohan Jung;Jina Kim;Kwang Suk Ko
    • Journal of Microbiology and Biotechnology
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    • v.32 no.11
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    • pp.1447-1453
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    • 2022
  • Prohibitin 1 (Phb1) is a pleiotropic protein, located mainly in the mitochondrial inner membrane and involved in the regulation of cell proliferation and the stabilization of mitochondrial protein. Acetaminophen (APAP) is one of the most commonly used over-the-counter analgesics worldwide. However, at high dose, the accumulation of N-acetyl-p-benzoquinone imine (NAPQI) can lead to APAP-induced hepatotoxicity. In this study, we sought to understand the regulation of mRNA expression in relation to APAP and GSH metabolism by Phb1 in normal mouse AML12 hepatocytes. We used two different Phb1 silencing levels: high-efficiency (HE, >90%) and low-efficiency (LE, 50-60%). In addition, the siRNA-transfected cells were further pretreated with 0.5 mM of Sadenosylmethionine (SAMe) for 24 h before treatment with APAP at different doses (1-2 mM) for 24 h. The expression of APAP metabolism-related and antioxidant genes such as Cyp2e1 and Ugt1a1 were increased during SAMe pretreatment. Moreover, SAMe increased intracellular GSH concentration and it was maintained after APAP treatment. To sum up, Phb1 silencing and APAP treatment impaired the metabolism of APAP in hepatocytes, and SAMe exerted a protective effect against hepatotoxicity by upregulating antioxidant genes.

EPS Production, PHB Accumulation and Abiotic Stress Endurance of Plant Growth Promoting Methylobacterium Strains Grown in a High Carbon Concentration

  • Woo, Sung-Man;Subramanian, Parthiban;Ramasamy, Krishnamoorthy;Joe, M. Melvin;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.4
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    • pp.572-581
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    • 2012
  • In this study, we compared growth pattern, floc yield, Exo-polysaccharides (EPS) production, Poly-${\beta}$-hydroxybutyrate (PHB) accumulation, resistance to osmotic and acid stress in Methylobacterium strains CBMB20, CBMB27, CBMB35, and CBMB110. Modified high C:N ratio medium denoted as HCN-AMS medium was used with a C:N ratio of 30:1. The HCN-AMS medium favored increased growth in all the studied strains. All Methylobacterium strains tested positive for EPS production and showed positive fluorescence with calcoflour stain. Elevated levels of EPS production from 4.2 to 75.0% was observed in HCN-AMS medium. Accumulation of PHB in HCN-AMS medium increased by 3.8, 36.7, and 12.0% in strains CBMB27, CBMB35, and CBMB110 respectively. Among the abiotic stresses, osmotic stress-induced growth inhibition of Methylobacterium strains was found to be lowered when grown in HCN-AMS medium. Likewise, growth inhibition due to acid stress at pH 5.0 was lower for strains grown in HCN-AMS medium compared to growth in AMS medium. Enhanced survivability under stress conditions may be attributed to the high EPS and PHB production at increased carbon concentration in the growth medium.

Determination of Tensile Modulus of PHB/PEN/PET Fiber Using Modified Halpin-Tsai Equation (변형 Halpin-Tsai식에 의한 PHB/PEN/PET 섬유의 탄성률 예측)

  • 정봉재;김성훈;이승구;전한용
    • Polymer(Korea)
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    • v.24 no.6
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    • pp.810-819
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    • 2000
  • Poly(p-hydroxybenzoate) (PHB)/poly(ethylene terephthalate) (PET) 8/2 thermotropic liquid crystalline copolyester, poly(ethylene 2,6-naphthalate) (PEN), and PET ternary blend was spun to fiber by melt spinninB process, and tensile properties of the fibers were measured. The matrix of the fibers, PET and PEN, were dissolved in ο-chlorophenol at 55$^{\circ}C$ for 2 hours, and the liquid crystalline polymer fibrils were observed using a scanning electron microscope. Halpin-Tsai equation for modulus calculation of short fiber reinforced composite and the rule of mixture for continuous reinforcement composite were modified, and the tensile modulus were calculated and compared with experimental modulus. To minimize difference between the theoretical and the experimental moduli, dimensionless viscosity constant (K) was given and used to modify two equations. The theoretical tensile modulus using the newly modified equations presentel a similar to the experimental tensile modulus of composite, and the modified equations presented a unique way to determine the tensile modulus of the liquid crystalline polymer reinforced thermoplastic composites.

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The effect of environmental factors affecting to the growth of Rhizobium japonicum (Rhizobium japonicum의 생장 및 poly$\beta$hydroxybutyric acid(PHB)의 축적에 미치는 환경요인의 영향)

  • 이기성;최영길
    • Korean Journal of Microbiology
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    • v.20 no.4
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    • pp.195-200
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    • 1982
  • Rhizobium japonicum was isolated from the nodule of soybean root grown at the reclaimed tidal land in Kang-Wha island. The effect of pH and salt concentration to the viability of the isolated strain were examined in relationship between microbial growth and accumulation of PHB. Optimal pH value for the good viability of the isolated strain was 7.0 and also, at 5.0 and 6.0 viability was favorable to large extent, but 9.0 was unfavorable. Examined the effect of salt concentration treated two times as of the salinity in the reclaimed tidal land, viability of the isolated strain showed about 30 to 40%. And also in treatment with NaCl(40g/l) whatever the pH value adopted, viability was mostly less than 10%. The amount of accumulated PHB was relatively high at low pH value(5-6) and at high salt concentrration, respectively.

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