• KSBB Journal
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• v.22 no.3
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• pp.146-150
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• 2007

Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] microspheres were prepared using solvent evaporation technique. P(3HB-co-4HB) with 3.9 mol% 4HB was synthesized by fed-batch culture of Ralstonia eutropha. The effects of concentration and type of surfactant (Tween 80, sodium dodecylsulfate, and polyvinyl alcohol), addition of dispersion stabilizer (Acacia), concentration of polymer and model drug (bovine serum albumin) on particle size of the microspheres and their in vitro drug release characteristics were investigated. The average particle size of the microspheres decreased with the addition of dispersion stabilizer and increased with the concentration of surfactant, drug and polymer. Amount of drug release increased with decreasing particle size of the microspheres.

• 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.

• Journal of Microbiology and Biotechnology
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• v.11 no.2
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• pp.333-336
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• 2001

An isolated phbC gene from Alcaligenes latus was reintroduced into the parent A. latus through the transformation process, and the effect of the amplified phbC gene on the biosynthesis of P(3-hydroxybutyrate-3-hydroxyvalerate) [P(3HB-3HV)] and P(3-hydroxybutyrate-4-hydroxybutyrate) [P(3HB-4HB)] in the transformant A. latus was investigated. The biosynthesis rate and content of the above copolymers increased up to 1.3-fold after enforcing its own phbC gene, and the molar fractions of 3HV and 4HB in P(3HB-3HV) and P(3HB-4HB) also changed remarkably from 35.0 to 48.0% and from 34.0 to 56.0%, respectively, showing a critical role of PHB synthase which catalyzes the polymerizing reactions between eiher 3HV or 4HB from precursor compounds and 3HB.

• KSBB Journal
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• v.19 no.5
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• pp.358-362
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• 2004

Characteristics of cell growth and poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] synthesis was investigated through flask and batch cultures of Alcaligenes latus and Comamonas acidovorans. The specific growth rate of C. acidovorans increased with yeast extract concentration and decreased with 1,4-butanediol concentration. Optimum glucose concentration for growth of C. acidovorans was 20 g/L. In one-step flask cultures of C. acidovorans, final dry cell weight and PHA content decreased with the ratio of 1,4-butanediol to glucose, while the 4HB fraction in copolymers gradually increased to 100 $mol\%$ with an initial 1,4-butanediol concentration of 20 g/L as single carbon source. The specific growth rate of A. latus decreased with v-butyrolactone concentration and optimum sucrose concentration for growth was 10 g/L. In batch cultures of A. latus, 4HB fraction increased with initial v-butyrolactone concentration. P(3HB-co-4HB) with 19 $mol\%$ 4HB was obtained when the initial ratio of v-butyloractone (g/L) to sucrose (g/L) was 10 : 10.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.603-606
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• 2005

Ralstonia eutropha NCIMB 11599 and ATCC 17699 were grown, and their productions of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] compared. In flask cultures of R. eutropha NCIMB 11599, cell concentration, P(3HB-co-4HB) concentration and polymer content decreased considerably with increases in the ${\gamma}-butyrolactone$ concentration, and the 4HB fraction was also very low (maximum 1.74 mol%). In fed-batch cultures of R. eutropha NCIMB 11599, glucose and ${\gamma}-butyrolactone$ were fed as the carbon sources, under a phosphate limitation strategy. When glucose was fed as the sole carbon source, with its concentration controlled using an on-line glucose analyzer, 86% of the P(3HB) homopolymer was obtained from 201g/L of cells. In a two-stage fed-batch culture, where the cell concentration was increased to 104g/L, with glucose fed in the first step and constant feeding of ${\gamma}-butyrolactone$, at 6g/h, in the second, final cell concentration at 67h was 106g/L, with a polymer content of 82%, while the 4HB fraction was only 0.7mol%. When the same feeding strategy was applied to the fedbatch culture of R. eutropha ATCC 17699, where the cell concentration was increased to 42 g/L, by feeding fructose in the first step and ${\gamma}-butyrolactone$ (1.5g/h) in the second, the final cell concentration, polymer content and 4HB fraction at 74h were 51g/L, 35% and 32 mol%, respectively. In summary, R. eutropha ATCC 17699 was better than R. eutropha NCIMB 11599 in terms of P(3HB-co-4HB) production with various 4HB fractions.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.722-725
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• 1999

Filamentation-suppressed recombinant Escherichia coli strain harboring the Alcaligenes latus polyhydroxyalkanoate (PHA) biosynthesis genes and the E. coli ftsZ gene was constructed and cultivated for the production of poly(3-hydroxybutyrate) [P(3HB)] with high concentration and high content. By the pH-stat fed-batch culture of this recombinant E. coli strain XL1-Blue(pJC5), the final cell concentration and P(3HB) concentration obtained in 44.25h were 172.2g cell dry weight/l and 141.9g P(3HB)/l, respectively, resulting in productivity of 3.21g P(3HB)/l-h. More importantly, the P(3HB) content obtained was 82.4 wt %, which was significantly higher than that obtained with the recombinant E. coli harboring only the PHA biosynthesis genes.

• Journal of Microbiology and Biotechnology
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• v.8 no.5
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• pp.540-542
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• 1998

Five microorganisms capable of degrading an aromatic medium-chain-length polyhydroxyalkanoate ($PHA_{MCL}$), poly(3-hydroxy-5-phenylvalerate) (PHPV), were isolated from wastewater-treatment sludge. Among the isolates, JS02 showed degrading activity consistantly during several transfers. The isolate JS02 could hydrolyze another aromatic MCL copolyester, poly(3-hydroxy-5-phenoxyvalerate-co-3-hydroxy-7-phenoxyheptanoate), ［P(5POHV-co-7POHH)］, and other short-chain-length PHAs ($PHA_{SCL}) such as poly(3-hydroxybutyrate) ［P3(HB)］, poly(3-hydroxybutyrate-co-4-hydroxybutyrate) ［P(3 HB-co-4 HB)］, and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) ［P(3HB-co-3HV)］ with relatively low activity. The culture supernatant of JS02 showed hydrolyzing activity for the p-nitrophenyl esters of fatty acids.

• Polymer(Korea)
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• v.29 no.1
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• pp.54-58
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• 2005

The hydrolytic behavior of microbial poly[(R)-3-hydroxybutyrate]](P(3HB)) has been studied by using two model systems, Langmuir monolayer and solution-grown single crystals (SCs), for elucidating the mechanism for both alkaline and enzymatic degradations. An initial degradation of SCs of P(3HB) leads to breakup lamellae parallel to their short axis (b-axis). Similarly, ridge formation on the lamellar surface appears along the b-axis at lower quenching temperature than melting temperature. Both results support that the lamellar crystals contain less-ordered and more thermally sensitive regions along the b-axis. Although the enzymatic hydrolysis of P(3HB) monolayers was similar to its alkaline one, the enzymatic degradation of P(3HB) monolayers occurred at higher constant surface pressure than the alkaline degradation. This behavior might be attributed to the size of enzymes which is much larger than that of alkaline ions; that is, the enzymes need larger contact area with monolayers to be activated.

• Journal of Korean Society of Environmental Engineers
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• v.35 no.2
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• pp.109-114
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• 2013

Polyhydroxyalkanoates (PHAs) are synthesized by numerous bacteria as carbon and energy storage compounds and are raw materials for biocompatible plastics. In this paper, the effect of $CO_2$ concentrations on the growth of C. necator and the accumulation of Poly(3-hydroxybutyrate) (P(3HB)) are investigated by increasing the $CO_2$ concentration in the substrate gas mixture. During 6 d cultivation in a nitrogen-present mineral medium, the $CO_2$ concentration did not affect the growth of the cells, while the Poly(3- hydroxybutyrate) (P(3HB)) content decreased with increasing $CO_2$ concentrations from 1% to 20%. During 4 d cultivation in the nitrogen-limited medium, the P(3HB) accumulation was the greatest at 3% $CO_2$; however, the total amount of accumulated P(3HB) was the greatest at 1% $CO_2$, which decreased with increasing $CO_2$ concentrations. The results indicate that the gas mixture with 1% $CO_2$ is the most effective in both growing the cells and accumulating P(3HB) under our experimental conditions.

• Microbiology and Biotechnology Letters
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• v.24 no.1
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• pp.92-100
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• 1996

For the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)(P(3HB-co-3HV)) from butyric acid and valeric acid, 10 strains of bacteria capable of producing P(3HB-co-3HV) were isolated from soil. Among them, the strain HJ-067 showed the best ability of producing P(3HB-co-3HV), and was indentified as a Azotobacter sp. For the production of P(3HB-co-3HV), the optimum concentrations of butyric and valeric acid were 3.0g/l, respectively. The most effective nitrogen source was $(NH_4)_{2}SO_4$ at an optimum concentration of 0.75g/l, which was equivalent to 21.36 in C/N ratio. Deficiency of the cationic metal ions ($Zn^{2+},\;Co^{2+},\;Mn^{2+}$) in the proguction medium had stimulating effect on P(3HB-co-3HV) accumulation, especially in the manganese. deficient medium. The optimum temperature for P(3HB-co-3HV) production was 27$^{\circ}C$ and the optimum initial pH was 7.0. Under the optimum conditions, 1.82g/l of P(3HB-co-3HV) and 3.00g/l of dry biomass were produced after 36 hour cultivation, and the P(3HB-co-3HV) yield and HV% were 60.60% (w/w), 15.92%, respectively.