• Journal of Applied Biological Chemistry
• /
• v.42 no.1
• /
• pp.1-6
• /
• 1999

Poly-${\beta}$-hydroxybutyrate (PHB) and enzymes related PHB metabolism have been measured in nitrogen-fixing symbiosis of chickpea and cowpea plants. Bacteroids from chickpea and cowpea contained PHB to 0.8% and 43% of their dry weight, respectively, whereas the free-living cells CC 1192 and I 16 produced $285{\pm}55mg$ and $157{\pm}18mg$ of PHB g (dry weight)$^{-1}$. To further understand why chickpea bacteroids contained little PHB, the enzyme activities of PHB metabolism (3-ketothiolase, acetoacetyl-CoA reductase, PHB depolymerase, and 3-hydroxybutyrate dehydrogenase), the TCA cycle (malate dehydrogenase, citrate synthase, and isocitrate dehydrogenase), and related reactions (malic enzyme, pyruvate dehydrogenase, and glutamate:2-oxoglutarate transaminase) were compared in extracts from chickpea and cowpea bacteroids and the respective free-living bacteria. Significant differences were observed between chickpea and cowpea bacteroids and between the bacteroid and free-living forms of CC 1192, with respect to the capacity for some of these reactions. It is indicated that a greater potential for oxidizing malate to oxaloacetate in chickpea bacteroids could be a factor that favors the utilization of acetyl-CoA in TCA cycle rather than for PHB synthesis.

• Microbiology and Biotechnology Letters
• /
• v.18 no.3
• /
• pp.273-279
• /
• 1990

For poly- $\beta$ -hydroxybutyrate (PHB) production, a pink-pigmented facultative methylotrophic bacterium (PPFM) P-10 was newly isolated from soils through methanol-enrichment culture. The optimal medium composition for cell growth was 1.0% (vlv) of methanol as carbon source and l.Og/l of ,TEX>$NH_4Cl$, equivalent to C/N ratio of 13.2 at pH 7.0 and $30^{\circ}C$. To investigate the optimal condition for YHB accumulation, two-stage culture technique was adopted; first stage for cell growth and second stage for accumulation of PHB providing unbalanced growth conditions. The optimal PHB accumulation was 1.0% (vIv) of methanol and 0.26gll of $NH_4Cl$, C/N of 50.8 at pH 6.0. To overcome methanol inhibition on cell growth, intermittent feeding fed-batch culture technique was employed, and the cell concentration as high as 14gll with 40% of PHB was achieved. The purified PHB was identified using IR and $1^H NMR$ as homopolymer of 8hydroxybutyric acid. The absorption spectrum of extracted pink colored microbial pigment was alsa investigated.

• KSBB Journal
• /
• v.10 no.2
• /
• pp.176-182
• /
• 1995

The environmental and physiological factors affecting the production of exopolysaccharide (Methylan) and Poly-${\beta}$-hydroxybutyrate(PHB) by Methylobacterium organophilum were investigated. The maximum PHB content was obtained at $38^{\circ}C$ whereas maximum polysaccharide concentration was $3.54g/\ell$ at $30^{\circ}C$. Optimum pH was pH 7-8 for PHB production and pH 6-7 for polysaccharide production, respectively. Under the condition of $Mo^{2+}, Mg^{2+} or Mn^{2+}$ limitation with nitrogenlimitation, the PHB accumulation was increased, whereas the polysaccharide production was decreased as compared with that of solenitrogenlimitation. Under the condition of sole K+ limitation, cell growth was significantly inhibited and no polysaccharide was produced. However, the PHB content was as high as 60% of dry cell weight. Effect of C/N ratios (methanol/ammonium) in the feeding solution was examined for the simultaneous production of polysaccharide and PHB. The higher ratio of C/N showed the lower cell growth, higher content of PHB in cells, and higher yield of polysaccharide.

• Journal of Microbiology and Biotechnology
• /
• v.11 no.2
• /
• pp.310-316
• /
• 2001

Poly-3-hydroxybutyrate (PHB) synthase catalyzed the last enzymic step to synthesize the intracellular PHB of Rhodobacter sphaeroides. No PHB was detected when the phbC coding for PhB synthase was interrupted, and its expression was regulated at the level of transcription. The cellular PHB content increased about four- to six-fold during the growth transition from the exponential to the early stationary phase under both aerobic and photoheterotrophic conditions. The PHB content during the aerobic growth seemed to be determined by the PhB synthase activity. However, the PHB synthase activity of photoheterotrophically grown cells did not correlate with the PhB content, suggesting a photoheterotrophic regulation different from the aerobic control. Thus, the PHB content of R. sphaeroides was regulated at the transcription level only under aerobic conditions.

• Korean Chemical Engineering Research
• /
• v.55 no.3
• /
• pp.363-368
• /
• 2017

Methylotrophy is able to use reduced one-carbon compound, such as methanol and methylamine, as a sole carbon source. Methylobacterium extorquens AM1 is the most extensively studied methylotroph utilizing serine-isocitrate lyase cycle. Because the Poly 3-hydroxybutyrate (PHB) synthesis pathway in M. extorquens AM1 is likely to interlink with EMCP (ethylmalonyl-CoA pathway), glyoxylate, and TCA cycles, regulation of PHB production is needed to produce EMCP-derived acid or TCA acids. To adjust carbon flux to PHB production, PhaR, which seems to have function of regulator of PHB synthesis and acetyl-CoA flux, was knocked out in M. extorquens AM1 by using markerless gene deletion methods. As a result, PHB granules were remarkably reduced in the knockout strain ${\Delta}phaR$ compared to parental strain. Although lag phase was extended for 12h, ${\Delta}phaR$ showed similar cell growth and methanol consumption rate compared to wild type.

• Journal of Microbiology and Biotechnology
• /
• v.33 no.8
• /
• pp.1076-1083
• /
• 2023

Poly(3-hydroxybutyrate) (PHB) is a biodegradable and biocompatible bioplastic. Effective PHB degradation in nutrient-poor environments is required for industrial and practical applications of PHB. To screen for PHB-degrading strains, PHB double-layer plates were prepared and three new Bacillus infantis species with PHB-degrading ability were isolated from the soil. In addition, phaZ and bdhA of all isolated B. infantis were confirmed using a Bacillus sp. universal primer set and established polymerase chain reaction conditions. To evaluate the effective PHB degradation ability under nutrient-deficient conditions, PHB film degradation was performed in mineral medium, resulting in a PHB degradation rate of 98.71% for B. infantis PD3, which was confirmed in 5 d. Physical changes in the degraded PHB films were analyzed. The decrease in molecular weight due to biodegradation was confirmed using gel permeation chromatography and surface erosion of the PHB film was observed using scanning electron microscopy. To the best of our knowledge, this is the first study on B. infantis showing its excellent PHB degradation ability and is expected to contribute to PHB commercialization and industrial composting.

• Journal of Microbiology and Biotechnology
• /
• v.13 no.3
• /
• pp.477-481
• /
• 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.

• 한국생물공학회:학술대회논문집
• /
• 2000.11a
• /
• pp.175-178
• /
• 2000

Biodegradability of the $poly({\beta}-hydroxybutyrate-co-3 -hydroxyvalerate)$ [PHB/V] containing 0, 10 and 15mol% hydroxyvalerate [HV] was studied. Bioderadability of PHB/V was evaluated at $30^{\circ}C$ for 58 days and $55^{\circ}C$ for 33 days by monitoring the time-dependent changes in weight loss(erosion) of aerobic conditions in a temperature-controlled microcosms containing the earthworm cast($30^{\circ}C$) and compost ($55^{\circ}C$). It was found that PHB/V biodegradability occurred with increasing HV monomer concentration from 0 mol% to 15 mol%.

• Korean Journal of Microbiology
• /
• v.36 no.4
• /
• pp.299-305
• /
• 2000

Degradation behavior of the three commercial biodegradable polymers, namely poly(3-hydroxybutyrate) (PHB) Sky-Green/sup R/ (SG) and Mater-Bi/sup R/ (MB) was investigated using bacteria isolated from activated sludge and farm soil. Three PHB degrading bacteria, three SG degrading bacteria and one MB degrading bacteria were isolated. The PHB degrading bacteria were identified to be Flavimonas oryzihabitans, Corynebacterium pseudodiphtheriticum and Micrococcus diversus, while Pseudomonas vesicuraris, Pasteurlla multocida and Flavobacterium odoratum were identified as SG degrading bacteria. As for MB, Pseudomonas vesicuraris was isolated. The shake flask test for 28 days indicated that the rate of biodegradation of PHB, SG and MB in terms of weight loss were about 44∼69% 25∼32% and 29% respectively. The surface morphology of PHB, SG andMB films before and after degradation by microorganisms in an activated sludge soil was observed under SEM, demonstrating that the film surface had a very porous structure, and that microorganisms colonized heavily on the film surface. TOC and pH variation as a result of abiotic hydrolysis, or microbial growth in the absence of the polymers were compared to those due to degradation by F. oryzihabitans. Abiotic hydrolysis of PHB was three times as fast as that of SG and MB. Addition of yeast extract to the basal liquid medium accelerated the biodegradation of the polymers. Biodegradation of PHB was always faster than that of SG and MB irrespectively of the presence of yeast extract in the basal liquid medium.

• Microbiology and Biotechnology Letters
• /
• v.24 no.3
• /
• pp.352-356
• /
• 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.