• KSBB Journal
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• v.25 no.3
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• pp.239-243
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• 2010

Six strains of Ralstonia eutropha were grown to investigate characteristics of polyhydroxyalkanoates (PHA) synthesis from vegetable oils or glycerol. Poly(3-hydroxybutyrate) homopolymer was formed using soybean oil, olive oil, or glycerol as carbon source, while poly(3-hydroxybutyrate-co-4-hydroxybutyrate) or poly(3-hydroxybutyrateco-3-hydroxyvalerate) copolymers were synthesized by co-feeding $\gamma$-butyrolactone or pentanoic acid, respectively. Optimum strain was determined as R. eutropha KCTC 2662 in terms of final cell concentration and PHA content. From 20 g/L of soybean oil (optimum substrate), cell concentration and PHA content at 72 h ranged 1.7~9.2 g/L and 70~92 wt%, respectively.

• KSBB Journal
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• v.29 no.4
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• pp.244-249
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• 2014

Biosynthesis pathway of medium-chain-length (MCL) polyhydroxyalkanoates (PHA) from fatty acid ${\beta}$-oxidation pathway was constructed in recombinant Escherichia coli by introducing the Pseudomonas sp. 61-3 PHA synthase gene (phaC2) and the maoC genes from Pseudomonas putida, Sinorhizobium meliloti, and Ralstonia eutropha. The metabolic link between fatty acid ${\beta}$-oxidation pathway and PHA biosynthesis pathway was constructed by MaoC, which is homologous to P. aeruginosa (R)-specific enoyl-CoA hydratase (PhaJ1). When the E. coli W3110 strains expressing the phaC2 gene and one of the maoC genes from P. putida, Sinorhizobium meliloti, and Ralstonia eutropha were cultured in LB medium containing 2 g/L of sodium decanoate as a carbon source, MCL-PHA that mainly consists of 3-hydroxyhexanoate (3HHx), 3-hydroxyoctanoate (3HO) and 3-hydroxydecanoate (3HD), was produced. The monomer composition of PHA and PHA contents varied depending on MaoC employed for the production of PHA. The highest PHA content of 18.7 wt% was achieved in recombinant E. coli W3110 expressing the phaC2 gene and the P. putida maoC gene. These results suggest that MCL-PHA biosynthesis pathway can be constructed in recombinant E. coli strains from the b-oxidation pathway by employing MaoC able to supply (R)-3-hydroxyacyl-CoA, the substrate of PHA synthase.

• Environmental Engineering Research
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• v.20 no.3
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• pp.246-253
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• 2015

The effects of carbon-to-nitrogen (C/N) ratio in simulated rice straw hydrolysates using glucose and ammonium chloride on polyhydroxyalkanoates (PHA) accumulation by Cupriavidus necator was investigated. In general, PHA accumulation rate was higher under higher degrees of N-deficient conditions (e.g., C/N ratio of 360:1) than lower degrees of N-deficient conditions (e.g., C/N ratio of 3.6:1 and 36:1). Also, the most PHA accumulation was observed during the first 12 h after the PHA accumulation initiation. This study showed that the similar PHA accumulation could be achieved by using different accumulation periods depending on C/N ratios. N source presence was important for new cell production, supported by approximately ten times greater PHA accumulation under the N-deficient condition ($NH_4Cl$ 0.01 g/L) than the N-free (without $NH_4Cl$) condition after 96 h. C/N ratio of the rice straw hydrolysate was approximately 160:1, based on the glucose content, and this accumulated $0.36{\pm}0.0033g/L$ PHA with PHA content of $21{\pm}3.1%$ after 12 h. Since external C or N source addition for C/N ratio adjustment increases production cost, an appropriate accumulation period may be used for PHA accumulation from organic wastes, based on the PHA accumulation patterns observed at various C/N ratios and C and N concentrations.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.123-127
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• 2004

Pseudomonas sp. MBEL21 was newly isolated from soil samples and found to accumulate medium-chain-length Polyhydroxyalkanoates(MCL-PHAs) using oleic acid as a sole carbon source. Among the various nutrient limiting conditions examined, including nitrogen, sulfur and phosphorus, only phosphorus limitation supported the accumulation of MCL-PHAs up to 15 wt％ of dry cell weight in flask cultures. MCL-PHAs produced by Pseudomonas sp. MBEL21 was mainly composed of 3-hydroxy-5-cis-tetradecenoate. Fed-batch culture of Pseudomonas sp. MBEL21 by novel feeding strategies based on cell growth charcteristics was carried out under phosphorus limitation using oleic acid as a sole carbon source. The final cell concentration and PHA content of 82 g/L and 28 wt％, respectively, were obtained. Furthermore, PHA consisted of MCL-hydroxyalkanoates and 3-hydroxybutyrate could be produced using olive oil as a sole carbon source.

• Journal of Microbiology
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• v.45 no.2
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• pp.87-97
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• 2007

Medium-chain-length polyhydroxyalkanoates (MCL-PHAs), which have constituents with a typical chain length of $C_{6}-C_{14}$, are polyesters that are synthesized and accumulated in a wide variety of Gram-negative bacteria, mainly pseudomonads. These biopolyesters are promising materials for various applications because they have useful mechanical properties and are biodegradable and biocompatible. The versatile metabolic capacity of some Pseudomonas spp. enables them to synthesize MCL-PHAs that contain various functional substituents; these MCL-PHAs are of great interest because these functional groups can improve the physical properties of the polymers, allowing the creation of tailor-made products. Moreover, some functional substituents can be modified by chemical reactions to obtain more useful groups that can extend the potential applications of MCL-PHAs as environmentally friendly polymers and functional biomaterials for use in biomedical fields. Although MCL-PHAs are water-insoluble, hydrophobic polymers, they can be degraded by microorganisms that produce extracellular MCL-PHA depolymerase. MCL-PHA-degraders are relatively uncommon in natural environments and, to date, only a limited number of MCL-PHA depolymerases have been investigated at the molecular level. All known MCL-PHA depolymerases share a highly significant similarity in amino acid sequences, as well as several enzymatic characteristics. This paper reviews recent advances in our knowledge of MCL-PHAs, with particular emphasis on the findings by our research group.

• Microbiology and Biotechnology Letters
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• v.51 no.3
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• pp.257-267
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• 2023

The potential polyhydroxyalkanoates (PHA)-producing bacteria, Bacillus megaterium PP-10, was successfully isolated and studied its feasibility for utilization of pineapple peel waste (PPW) as a cheap carbon substrate. The PPW was pretreated with 1% (v/v) H₂SO₄ under steam sterilization and about 26.4 g/l of total reducing sugar (TRS) in pineapple peel hydrolysate (PPH) was generated and main fermentable sugars were glucose and fructose. A maximum cell growth and PHA concentration of 3.63 ± 0.07 g/l and 1.98 ± 0.09 g/l (about 54.58 ± 2.39%DCW) were received in only 12 h when grown in PPH. Interestingly, PHA productivity and biomass yield (Y_x/s) in PPH was about 4 times and 1.5 times higher than in glucose. To achieve the highest DCW and PHA production, the optimal culture conditions e.g. carbon to nitrogen ratios of 40 mole/mole, incubation temperature at 35℃ and shaking speed of 200 rpm were performed and a maximum DCW up to 4.24 ± 0.04 g/l and PHA concentration of 2.68 ± 0.02 g/l (61% DCW) were obtained. The produced PHA was further examined its monomer composition and found to contain only 3-hydroxybutyrate (3HB). This finding corresponded with the presence of class IV PHA synthase gene. Finally, certain thermal properties of the produced PHA i.e. the melting temperature (T_m) and the glass transition temperature (T_g) were about 176℃ and -4℃, respectively whereas the M_w was about 1.07 KDa ; therefore, the newly isolated B. megaterium PP-10 is a promising bacterial candidate for the efficient conversion of low-cost PPH to PHA.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.207-210
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• 2000

Pseudomonas oleovorans was cultivated to produce medium chain length polyhydroxyalkanoates (MCL-PHA) fram octanoic acid and ammonium nitrate as carbon and nitrogen source, respectively, by a pH-stat fed-batch culture technique. The octanoate concentration of the culture broth was maintained below 4 g/L by feeding the mixture of octanoic acid and ammonium nitrate when the culture pH rose above high limit. The effect of the ratio of octanoic acid to ammonium nitrate (C/N ratio) in the feed on the PHA production was examined. The final cell concentrations of 62.5, 54.7, and 9.5 g/L, PHA contents of 62.9, 75.1, and 67.6% of dry cell weight, and productivities of 1.03, 0.632, and 0.161 g/L/h were obtained when the C/N ratio in the feed were 10, 20, and 100 g octanoic acid/g ammonium nitrate, respectively.

• Journal of Microbiology and Biotechnology
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• v.31 no.2
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• pp.338-347
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• 2021

Polyhydroxyalkanoates (PHA) are a family of microbial polyesters that are used as biodegradable plastics in replacement of conventional plastics for various applications. However, the high production cost is the barrier for PHA market expansion. This study aimed to utilize food waste as low-cost feedstock to produce poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) by Haloferax mediterranei. The effects of acetate (Ac), propionate (Pr), butyrate (Bu), and the short-chain carboxylates derived from food waste were examined on the microbial growth and PHBV production. Results showed that a mixture of carboxylates provided a 55% higher PHBV yield than glucose. The food-waste-derived nutrients achieved the yields of 0.41 to 0.54 g PHBV/g Ac from initial loadings of 450 mg/l to 1,800 mg/l Ac of total carboxylates. And the consumption of individual carboxylate varied between different compositions of the carbon source. The present study demonstrates the potential of using food waste as feedstock to produce PHBV by Haloferax mediterranei, which can provide economic benefits to the current PHA industry. Meanwhile, it will also help promote organic waste reduction in landfills and waste management in general.

• Journal of Life Science
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• v.33 no.8
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• pp.680-691
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• 2023

Peak oil, climate change, and microplastics caused by the production and usage of petroleum-based plastics have threatened the sustainability of our daily life, and this has emerged as a recent global issue. To solve this global issue, the production and usage of biodegradable eco-friendly bioplastics such as polyhydroxyalkanoates (PHAs) has been suggested as an alternative. Therefore, in this review, the present status of global PHA manufacturers, the advantages of the production of PHAs using marine-origin microorganisms (with their productivity potential) and further required research and development strategies for cost-competitive production of PHAs using marine-based microorganisms were investigated. In this review, PHAs produced from marine microorganisms were found to have similar physical properties to petroleum-based plastics but with several advantages that can reduce the costs of PHA production. Those advantages include, seawater used in the medium preparation step, and osmotic-based cell lysis technology used in the separation and purification steps. However, the PHA productivities from marine microorganisms showed somewhat lower efficiencies than those from the commercial strains isolated from terrestrial environments. In order to solve the problem, further research strategies using synthetic microbiology-based technology, the development of long-term continuous culture technology, and solutions to improve PHA efficiency are required to meet future market demands for alternative bioplastics.

• KSBB Journal
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• v.19 no.4
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• pp.331-334
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• 2004

In vivo characterization of FadB homologous enzymes including PaaG, YdbU and YgfG for medium-chain-length (MCL) polyhydroxyalkanoate (PHA) biosynthesis was carried out in fadB mutant Escherichia coli. Previously, it was reported that amplification of FadB homologous enzymes such as PaaG and YdbU in fadB mutant E. coli resulted in enhanced biosynthesis of MCL-PHA by greater than two fold compared with control strain. In this study, we constructed paaG fadB double mutant E. coli WB114 and ydbU fadB double mutant E. coli WB115 to investigate the roles of PaaG and YdbU in biosynthesis of MCL-PHA. Inactivation of paaG and ydbU genes in fadB mutant E. coli harboring Pseudomonas sp. 61-3 phaC2 gene reduced the MCL-PHA production to 0.16 and 0.16 PHA g/L, respectively from 2 g/L of sodium decanoate, which are much lower than 0.43 PHA g/L obtained with fadB mutant E. coli WB101 harboring the phaC2 gene. Also, we identified new FadB homologous enzyme YgfG, and examined its roles by overexpression of ygfG and construction of ygfG fadB double mutant E. coli WB113.