• KSBB Journal
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• v.18 no.5
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• pp.404-407
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• 2003

Two recombinant Escherichia coli strains, GCSC6576 harboring a plasmid pSYL107 containing the Ralstonia eutropha polyhydroxyalkanoate (PHA) biosynthesis genes and a fadR atoC mutant LS5218 harboring a plasmid pJC4 containing the Alcaligenes latus PHA biosynthesis genes were compared for their ability to synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV) from whey. The 3HV fraction could be increased by acetic acid induction and oleic acid supplementation in flask cultures of recombinant E. coli GCSC6576. With the pH-stat fed-batch culture of recombinant E. coli LS5218, we obtained a cell concentration, a P(3HB-co-3HV) concentration, a P(3HB-co-3HV) content, and a 3HV fraction of 31.8 g/L, 10.6 g/L, 33.4%, and 6.26 mol%, respectively in 39 h.

• 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.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.540-545
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• 2005

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3HB-co-3HV), copolyesters, with 3-hydroxyvalerate (3HV) contents ranging from 17 to 60 mol%, were produced by Alcaligenes sp. MT-16, and their biocompatibility evaluated by the growth of Chinese hamster ovary (CHO) cells and the adsorption of blood proteins and platelets onto their film surfaces. The number of CHO cells that adhered to and grew on these films was higher with increasing 3HV content. In contrast, the tendency for blood proteins and platelets to adhere to the copolyester surfaces significantly decreased with increasing 3HV content. Examination of the surface morphology using atomic force microscopy revealed that the surface roughness was an important factor in determining the biocompatibility of theses copolyesters. The results obtained in this study suggest that poly(3HB-co-3HV) copolyesters, with >30 mol% 3HV, may be useful in biocompatible biomedical applications.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.321-324
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• 2001

Two recombinant Escherichia coli strains, GCSC6576 harboring a plasmid pSYL107 containing the Ralstonia eutropha polyhyclroxyalkanoate (PHA) biosynthesis genes and a fadR atoC mutant LS5218 harboring a plasmid pJC4 containing the Alcaligenes latus PHA biosynthesis genes were compared for their ability to synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] from whey as the sale carbon source. With the pH-stat fed-batch culture of E. coli LS5218, 、 ,ve obtained a cell concentration, a P(3HB-co-3HV) concentration. a P(3HB-co-3HV) content, and a 3HV fraction of 31.8 g/L, 10.6 g/L, 33.4 wt%. and 6.26 mol%, respectively at 39 h.

• Korean Journal of Microbiology
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• v.34 no.4
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• pp.219-224
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• 1998

The cyclic metabolism of poly(3-hydroxyhutyrate-co-3-hydroxyvalerate) synthesized from glucose by Alcaligenes sp. SH-69 in the presence or absence of new carbon substrate was investigated. In batch culture, the content and weight average molecular weight of the copolymer already stored in the cell decreased rapidly when there was no other carbon source available. After the depletion of carbon source, the amount of high molecular weight copolymer decreased more rapidly than that of low molecular weight copolymer, and as a result, average molecular weight distribution shifted to the lower value. The addition of a mixture of glucose and levulinic acid when the initial carbon substrate, glucose, was nearly depleted supported the continual increase in cell mass and the accumulation of poly(3HB-co-3HV) with high molar fraction of 3HV. However, solvent fractionation of the polymer with acetone revealed the degradation of pre-existing polyhydroxyalkanoale (PHA) in parallel with the synthesis of PHA from new carbon substrate. Even though PHAs obtained from each substrate alone were the copolymer of 3HB and 3HV, it was found that the polymer accumulated in the cells grown by sequential feeding was mainly physical mixture of two poly(3HB-co-3HV) copolymers containing different molar fractions of 3HV.

• Microbiology and Biotechnology Letters
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• v.19 no.1
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• pp.94-99
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• 1991

- The further study for the identification of the previously reported pink-pigmented facultative methylotrophic bacterium (PPFM) GL-10 was carried out. The PPFM GL-10 was Gram nagative, rod, and motile by a single polarly inserted flagellum. The colonies were smooth, pink, circular, along with convex with entire margin. The isolate could utilize C1 compounds and a variety of multicarbon substrates as sole carbon and energy source. The isolate was obligately aerobic, and exhibited both catalase and oxidase activities. The deoxyribonucleic acid base composition was 65-67 mol% guanine plus cytosine. The isolate was mostly identical with Methylobacterium extorquens and named Methylobacterium sp. strain GL-10. Methylobacterium GL-10 accumulated a copolyester of 3-hydroxybutyrate and 3-hydroxyvalerate (poly-3HB/3 HV) when grown in nitrogen-free culture media containing sodium propionate as substrate at the second polyester accumulation stage. The composition of copolyester, as determined from $^1h$ NMR spectra, was 23 mol% of 3-hydroxyvalerate (3HV).

• Korean Journal of Microbiology
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• v.36 no.2
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• pp.155-160
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• 2000

Pseudomnas sp. HJ-2 is capable of producing a rubber-elastic polyhydroxyalkanoate (PHA) consisting of 3- hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), and 3-hydroxyheptanoate (3HHp) from heptanoic acid as the sole carbon source. The polyester produced was a blend of poly(3HB-co-3HV) and poly(3HHp). Although the mixing of poly(3HHp) fraction to poly(3HB-co-3HV) resulted in a decrease of modulus, the sole fraction of poly(3HB-co-3HV) with a high molar fraction of 3HV was shown to be an elastomer with the maximum percent strain of 740%. The biomass yield and the PHA synthesis were relatively high when the initial heptanoic acid concentration was 40 mM, and were significantly decreased when the substrate concentration exceeded 50 mM. The accumulation of PHA was stimulated by deficiency of nitrogen and phosphorus in the medium. The PHA contents and its monomeric compositions were greatly affected by pH and oxygen transfer rate. At pH 7.5, poly(3HB-~0.38% 3HV) was produced from heptanoic acid and a mixture of 95% 3HHp and 5% 3HV was produced at pH 8.0. Increased conten1 of 3HHp in the polyesters with lhe increasing oxygen transfer rate by agitation speed a1 a fixed aeration rate was observed.

• Journal of Microbiology
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• v.42 no.4
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• pp.346-352
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• 2004

Poly(3-Hydroxybutyrate-co­3-Hydroxyvalerate), poly(3HB-co-3HV), copolyesters with a variety of 3HV contents (ranging from 17 to $60\;mol\%$) were produced by Alcaligenes sp. MT-16 grown on a medium containing glucose and levulinic acid in various ratios, and the effects of hydrophilicity and crystallinity on the degradability of the copolyesters were evaluated. Measurements of thermo-mechanical pro­perties and Fourier-transform infrared spectroscopy in the attenuated total reflectance revealed that the hydrophilicity and crystallinity of poly(3HB-co-3HV) copolyesters decreased as 3HV content in the copolyester increased. When the prepared copolyester film samples were non-enzymatically hydrolysed in 0.01 N NaOH solution, the weights of all samples were found to have undergone no changes over a period of 20 weeks. In contrast, the copolyester film samples were degraded by the action of extra­cellular polyhydroxybutyrate depolymerase from Emericellopsis minima W2. The overall rate of weight loss was higher in the films containing higher amounts of 3HV, suggesting that the enzymatic degra­dation of the copolyester is more dependent on the crystallinity of the copolyester than on its hydro­philicity. Our results suggest that the degradability characteristics of poly(3HB-co-3HV) copolyesters, as well as their thermo-mechanical properties, are greatly influenced by the 3HV content in the copoly­esters.

• Journal of Microbiology
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• v.35 no.2
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• pp.127-133
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• 1997

Biosynthesis of polyhydroxybutyrate and copolymer consisting of 3-hydroxybutyrate and 3-hydroxyvalerate [poly(3HB-co-3HV)] by Bacillus thuringiensis R-510 grown with glucose or with mixtures of glucose and propionate was investigated. n-Alkanoic acids other than propionate were not precursors of 3HV units. The fraction of 3HV unit in the copolymer increased from 0 to 84 mol% of 3HV. Polymer yield decreased as the fraction of propionate was increased but the molecular weight distribution was not affected by the composition of carbon substrate. The minimum melting temperature (around 65.deg.C) of poly (3HB-co-3HV) copolymers was observed for the polymer bearing approximately 35 mol% of 3HV. Polyhydroxyalkanoates production by this organism was not dependent on nutritional limitation, but remarkably influenced by dissolved oxygen concentration in the culture medium. Low level of dissolved oxygen concentration prevented spore formation in the cells and stimulated the synthesis of polyhydroxyalkanoate. The composition of poly (3HB-co-3HV) produced by B. thuringiensis R-510 lyhydroxyalkanoate. The composition of poly(3HB-co-3HV) propduced by B. thuringiensis R-510 varied according to the growth time. However, there was no evidence that polymers isolated from cells were mixtures of immiscible polymers.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.6
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• pp.371-374
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• 2002

Production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)［P(3HB/V)］, by fed-batch culture of recombinant Escherichia coli harboring a plasmid containing the Alcaligenes latus polyhy-droxyalkanoate (PHA) biosynthesis genes, was examined in two pilot-scale fermentors with air supply only, In a 30 L fermentor having a XLa value of 0.11 S­$^1$, the final P(3HB/V) concentration and the P(3HB/V) content obtained were 29.6 g/L and 70.1 wt%, respectively giving a productivity of 1.37 g P(3HB/V)/L-h. In a 300 L fermentor having a XLa of 0.03 S­$^1$, the P(3HB/V) concentration and the P(3HB/V) content were 20.4 g/L and 69 wt%, respectively giving a productivity of 1.06g P(3HB/V)/L-h. These results suggest that economical production of P(3HB/V) is possible by fed-batch culture of recombinant E. coli in a large-scale fermentor having low KLa value.