• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.735-738
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• 2001

Metabolically engineered Escherichia coli strains harboring a plasmid containing a novel artificial polyhydroxyalkanoate (PHA) operon consisting of the Aeromonas PHA biosynthesis related genes and Ralstonia eutropha reductase gene were developed for the production of poly(3-hydroxybutyrate-co-hydroxyhexanoate) [P(3HB-co-3HHx)] from dodecanoic acid. By applying stepwise reduction of dissolved oxygen concentration (DOC) during the fermentation, the final dry cell weight, PHA concentration, and PHA content of 79 g/L, 21.5 g/L, and 27.2 wt%, respectively, were obtained in 40.8 h, which resulted in the PHA productivity of 0.53 g/L/h. The 3HHx fraction slowly increased during the fed-batch culture to reach a final value of 10.8 mol%. The 3HHx fraction in the copolymer could be increased by three fold when the Aeromonas hydrophila orfl gene was co-expressed with the PHA biosynthesis genes.

• Polymer(Korea)
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• v.31 no.3
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• pp.233-238
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• 2007

The purpose of the study is to apply composites of poly (glycolic acid) (PGA) with [poly(R) 3-hydroxybutyrate] (P3HB) or poly (butylenes succinate- co-L-lactate) (PBSL) as medical resorbable composite materials with the complement of hydrolysis rate of each component. As a result, it was confirmed that the PBSL/PGA and P3HB/PGA composite fiber were hydrolyzed in phosphate buffer solution. Also, it has been revealed that the degradation of PBSL/PGA are accelerated due to PGA producing glycolic acid which can act as a catalyst. In addition, the hydrolysis of PBSL/PGA was found to be accelerated by the presence of lipase PS. When the PBSL/PGA composite fiber was placed in the air, not much hydrolysis has proceeded. Also, it was confirmed that the P3HB/PGA composite fiber maintained proper tensile strength in the air. Therefore, these complex fibers can be adapted to use as environmentally suitable, medically absorbable composite materials.

• Polymer(Korea)
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• v.25 no.3
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• pp.334-342
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• 2001

Gentamicin sulfate (GS)-loaded poly(3-hydroxybutyrate-co-3-hydroxyvalerate)(PHBV) devices were prepared for controlled-release of antibiotics. In this study, the effects of thickness, hydroxyvalerate (HV) content, initial drug-loading ratio, and additive content on the release profile have been investigated. The morphology of devices was examined with scanning electron microscope (SEM) before and after in vitro release; their highly porous surface and cross-sectional were observed. It could be suggested that device would be affected by the packing of the HV and additive content, which would depend on their structure. A high performance liquid chromatography (HPLC) was used to detect and quantify the release of GS from the device. The drug release from all the devices showed biphasic release patterns, and some matrices released the incorporated antibiotic throughout 30 days with a near zero-order release rate. The release patterns were shown to be changed by altering the thickness, copolymer ratio, and additive content.

• Proceedings of the Korean Fiber Society Conference
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• 2001.10a
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• pp.199-202
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• 2001

현재 플라스틱에 의한 환경오염이 사회문제로까지 확대되면서 이를 해결하기 위한 한가지 대책으로 분해성 플라스틱의 개발이 절실히 요구되어 전세계적으로 많은 연구가 이루어지고 있다. 생분해성을 가지는 미생물 폴리에스테르 중에서 가장 대표적인 poly(3-hydroxybutyrate), PHB는 우수한 역학적 성질을 가지며, 또한 천연고분자 중에서 융점을 가지는 유일한 열가소성 재료이다. (중략)

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.731-734
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• 2001

An efficient fermentation strategy for the high level production of ultra-high-molecular weight poly(3-hdyroxybutyrate) (PHB) was developed. Although the cell and PHA concentrations obtained by flask cultures at different initial pH (6.0 or 6.9) were almost same level, the molecular mass of PHB produced were quite different along with the initial pH. When a recombinant Escherichia coli XL1-Blue harboring pJC2 containing the Alcaligenes latus PHB biosynthesis genes was cultivated in flask culture (pH 6.0), the PHB having a very high molecular weight of 22 MDa could be produced while only below 1 MDa at initial pH 6.9. The ultra-high-molecular weight PHB could be synthesized to high concentration of 89.8 g/L resulting in the PHB productivity of 2.07 g/L-h by simple fed-batch culture. In this study, we report that PHB having various molecular mass can be produced by employing metabolically engineered E. coli strains harboring the plasmids of different copy numbers containing the A. latus phbCAB genes.

• Journal of Microbiology and Biotechnology
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• v.32 no.1
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• pp.27-36
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• 2022

Ever since bioplastics were globally introduced to a wide range of industries, the disposal of used products made with bioplastics has become an issue inseparable from their application. Unlike petroleum-based plastics, bioplastics can be completely decomposed into water and carbon dioxide by microorganisms in a relatively short time, which is an advantage. However, there is little information on the specific degraders and accelerating factors for biodegradation. To elucidate a new strain for biodegrading poly-3-hydroxybutyrate (PHB), we screened out one PHB-degrading bacterium, Microbulbifer sp. SOL03, which is the first reported strain from the Microbulbifer genus to show PHB degradation activity, although Microbulbifer species are known to be complex carbohydrate degraders found in high-salt environments. In this study, we evaluated its biodegradability using solid- and liquid-based methods in addition to examining the changes in physical properties throughout the biodegradation process. Furthermore, we established the optimal conditions for biodegradation with respect to temperature, salt concentration, and additional carbon and nitrogen sources; accordingly, a temperature of 37℃ with the addition of 3% NaCl without additional carbon sources, was determined to be optimal. In summary, we found that Microbulbifer sp. SOL03 showed a PHB degradation yield of almost 97% after 10 days. To the best of our knowledge, this is the first study to investigate the potent bioplastic degradation activity of Microbulbifer sp., and we believe that it can contribute to the development of bioplastics from application to disposal.

• 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.

• KSBB Journal
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• v.29 no.6
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• pp.443-447
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• 2014

Biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) was examined in recombinant Escherichia coli W strain from sucrose. The Pseudomonas sp. MBEL 6-19 phaC1437 gene and the Clostridium propionicum pct540 gene, which encode engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 ($PhaC1_{Ps6-19}$) and engineered C. propionicum propionyl-CoA transferase ($Pct_{Cp}$), respectively, were expressed in E. coli W to construct key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli W expressing the phaC1437 gene and the pct540 gene could synthesize P(3HB-co-13mol%LA) up to the polymer content of 31.3 wt% when it was cultured in chemically defined MR medium containing 20 g/L of sucrose and 2 g/L of sodium 3-hydroxybutyrate. When Ralstonia eutropha phaAB genes were additionally expressed to provide 3-hydroxybutyrate-CoA (3HB-CoA) from sucrose, P(3HB-co-16mol%LA) could be synthesized from sucrose as a sole carbon source without supplement of sodium 3-hydroxybutyrate in culture medium, but the PHA content was decreased to 12.2 wt%. The molecular weight of P(3HB-co-16 mol%LA) synthesized in E. coli W using sucrose as carbon source were $1.53{\times}10^4$ ($M_n$) and $2.78{\times}10^4$ ($M_w$), respectively, which are not different from those that have previously been reported by other recombinant E. coli strains. Engineered E. coli strains developed in this study should be useful for the production of lactate-containing PHAs from sucrose, one of the most abundant and least expensive carbon sources.

• Korean Journal of Microbiology
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• v.39 no.2
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• pp.76-82
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• 2003

To prepare evolved PHB depolymerase with increased activity for PHB or P(3HB-co-3HV) compared to the activity of the original PHB depolymerase from Alcaligenes faecalis T1, random mutation of the cloned PHB depolymerase gene was performed by using a DNA shuffling method. A library of mutated PHB depolymerase genes from A. faecalis T1 was fused to the ice nucleation protein (INP) gene from Pseudomonas syringae in pJHCl 1 and approximately 7,000 transformants were isolated. Using M9 minimal medium containing PHB or P(3HB-co-3HV) as the carbon source, mutants showing alteration in PHB depolymerase activity were selected from the transformants. The PHB depolymease activity of the transformants was confirmed by the formation of halo around colony and the turbidity decrease tests using culture supermatants. The catalytic activity of PHB depolymerase of the best mutant II-4 for PHB or P(3HB-co-13 mol% 3HV) was approximately 1.8-fold and 3.2-fold, respectively, higher than that of the original PHB depolymerase. DNA sequence analysis revealed that three amino acid residues (Ala209Val, Leu258Phe, and Asp263Thr) were substituted in II-4. From the mutational analysis, it was presumed that the substitution of amino acids near catalytic triad to more hydrophobic amino acids enhance the catalytic activity of PHB depolymerase from A. faecalis T1.

• Journal of Microbiology and Biotechnology
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• v.7 no.6
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• pp.391-396
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• 1997

Two series of carbon sources, linear primary $C_1$~$C_9$ alcohols and linear $C_2$~$C_{10}$ monocarboxylic acids were tested for PHA synthesis in Paracoccus denitrificans. The results showed that the growth-associated synthesis of PHA could be referred only to the carbon sources with odd number of carbon except methanol. For all carbon sources with even number of carbon, nitrogen limitation was required to induce PHA synthesis in P. denitrificans. Poly(3-hydroxyvalerate)[P(3HV)] homopolymer was synthesized from $C_5$, $C_7$, and $C_9$ while growing in the presence of nitrogen, but the nitrogen depletion in the later growth period incorporated 3-hydroxybutyrate(3HB) unit into the polymer chain. The optimum C/N ratio for P(3HV) homopolymer production was found to be 10 when the strain was grown on 10 ml/l of valeric acid for 96 h. P. denitrificans synthesized P(3HB-co-3HV) copolymers from n-hexanoic and n-octanoic acid. The microstructural characterics of the P(3HB-co-3HV) copolymer from n-propanol was investigated using $^13C$-nuclear magnetic resonance spectroscopy, showing a structural heterogeneity.