• Polymer(Korea)
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• v.27 no.6
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• pp.542-548
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• 2003

Nanoparticles with unsaturated poly(hydroxyalkanoate)s (UPHAs) biosynthesized with Pseudo-monas oleovorans were prepared by spontaneous emulsification solvent diffusion method. The influence of nanoparticle formation was investigated with various experimental parameters such as sonication conditions, sol-vent, surfactant and polymer contents, etc. The physical and chemical properties of UPHAS and its nanoparticles were characterized using $^1$H- and $\^$13/C-nuclear magnetic resonance spectroscopies, attenuated total reflection infrared spectroscopy, differential scanning calorimetry and gel permeation chromatography. The morphology of particles was observed using scanning electron microscope and the size and distribution of nanoparticles were measured with electrophoretic light scattering spectrophotometer. The mean diameter of particles decreased with increasing sonication amplitude and time. The addition of ethanol into UPHAS chloroform solution decreased the particle size presumably due to increased solvent diffusion into water phase. The particle size increased with increased the concentration of UPHAS solution. Under the 2-4％ poly(vinyl alcohol) (PVA) aqueous solution the minimum mean diameter of particles was shown. The higher degree of hydrolysis and degree of polymerization of PVA increased the mean diameter of particles.

• Journal of the Korea Organic Resources Recycling Association
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• v.9 no.3
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• pp.119-126
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• 2001

A process for the production of bioplastics from wastewater with an open microbial culture was developed and evaluated. The process consists of a selection reactor to select bacteria in feast/famine regime and an accumulation reactor to produce PHA using selected bacteria. Polyhydroxyalkanoate(PHAs) accumulating bacteria could be efficiently grown in a sequencing batch reactor(SBR) without any growth limitation. For the high production of PHA limitation such as oxygen and nutrients seemed to be needed. Accumulation experiments were performed to evaluate the level of accumulation of PHA. Limited aeration had no effect, but nutrients limitation showed high accumulation. Bacteria which were selected in the SBR could accumulate PHA till 60% of cellular dry weight in accumulation experiments under nitrogen limitation. PHA accumulation rate decreased with increasing PHA content in the cells. Clearly, PHA accumulation rate has a strong correlation with the PHA content of the cells.

• Journal of Microbiology and Biotechnology
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• v.24 no.1
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• pp.59-69
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• 2014

One of the major challenges in metabolic engineering for enhanced synthesis of value-added chemicals is to design and develop new strains that can be translated into well-controlled fermentation processes using bioreactors. The aim of this study was to assess the influence of various fed-batch strategies in the performance of metabolically engineered Pseudomonas putida strains, ${\Delta}gcd$ and ${\Delta}gcd-pgl$, for improving production of medium-chain-length polyhydroxyalkanoates (mcl-PHAs) using glucose as the only carbon source. First we developed a fed-batch process that comprised an initial phase of biomass accumulation based on an exponential feeding carbon-limited strategy. For the mcl-PHA accumulation stage, three induction techniques were tested under nitrogen limitation. The substrate-pulse feeding was more efficient than the constant-feeding approach to promote the accumulation of the desirable product. Nonetheless, the most efficient approach for maximum PHA synthesis was the application of a dissolved-oxygen-stat feeding strategy (DO-stat), where P. putida ${\Delta}gcd$ mutant strain showed a final PHA content and specific PHA productivity of 67% and $0.83g{\cdot}l^{-1}{\cdot}h^{-1}$, respectively. To our knowledge, this mcl-PHA titer is the highest value that has been ever reported using glucose as the sole carbon and energy source. Our results also highlighted the effect of different fed-batch strategies upon the extent of realization of the intended metabolic modification of the mutant strains.

• 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.

• Microbiology and Biotechnology Letters
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• v.52 no.1
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• pp.102-104
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• 2024

Massilia sp. KACC 81254BP, isolated from a landfill on Jeju Island, Republic of Korea, possesses the capability to degrade polyhydroxyalkanoates (PHAs). The genomic analysis of strain KACC 81254BP consists of a circular chromosome comprising 5,028,452 base pairs with a DNA G+C content of 64.6%. This complete genome consists of a total of 4,513 genes, including those encoding the PHA degradation enzyme (PhaZ). This study offers valuable genomic insights into the enzymes responsible for degrading polyester plastics.

• Korean Journal of Microbiology
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• v.52 no.4
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• pp.455-462
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• 2016

Pseudomonas aeruginosa P-5 is an unusual organism capable of synthesizing polyhydroxyalkanoates (PHAs) consisting of 3-hydroxyvalerate (3HV) and medium-chain-length (MCL) 3-hydroxyalkanoate (3HA) monomer units when C-odd alkanoic acids are fed as the sole carbon source. Evaluation of the substrate chain-length specificity of two P. aeruginosa P-5 PHA synthases ($PhaC1_{P-5}$ and $PhaC2_{P-5}$) by heterologous expression of $PhaC1_{P-5}$ and $PhaC2_{P-5}$ genes in Pseudomonas putida GPp104 revealed that $PhaC2_{P-5}$ incorporates both 3HV and MCL 3HAs into PHA, whereas $PhaC1_{P-5}$ favors only MCL 3HAs for polymerization. In order to obtain $PhaC2_{P-5}$ mutants with altered substrate specificity, site-specific mutagenesis for $PhaC2_{P-5}$ was conducted. Amino acid substitutions of $PhaC2_{P-5}$ at two positions (Ser326Thr and Gln482Lys) were very effective for synthesizing copolymers with a higher 3HV fraction. When recombinant P. putida GPp104 harboring double mutated $phaC2_{P-5}$ gene ($phaC2_{P-5}QKST$) was grown on nonanoic acid, 2.5-fold increase of copolymer content with 3.8-fold increase of 3HV fraction was observed. The $phaC2_{P-5}QKST$-containing Ralstonia eutropha PHB-4 supplemented with valeric acid also produced copolymers consisting of 3HV and 3-hydroxyheptanoate with a high 3HV fraction. These results suggest that recombinants containing $phaC2_{P-5}QKST$ could be useful for production of new PHA copolymers with improved material properties.

• Journal of the Korea Organic Resources Recycling Association
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• v.14 no.1
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• pp.160-168
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• 2006

This study was performed to evaluate the effect of the addition of vegetable oils on Fenton treatment of PAHs-contaminated soil. Fenton reaction can be proceeded in the soils contaminated with PHAs only in the presence of $H_2O_2$ because of Fe content in the soil. In this case, optimum $H_2O_2$ concentration was 3%. When 17.5 mM $FeSO_4$(III) was added with 3% $H_2O_2$, the removal rate was increased up to 25%, whereas 19% of PAHs was removed with $H_2O_2$ alone. The addition of 1% of olive oil to the contaminated soil before the Fenton reaction or simultaneously increased the removal rate about 15%, compared to the case of Fenton reaction only. There were no significant differences in the removal rates of PAHs, regardless of different kinds and concentration of oils. (olive oil, soybean oil, and used-vegetable oil). The used-vegetable oils were not different from the new, expensive oils in the removal rate, so their use will be desirable in saving the money. In addition of 1% of olive oil after the reaction of 3% $H_2O_2$ and 2.5 mM $FeSO_4$(III), the removal rates of 3~4 and 5~6 ring compounds were increased 13% and 17%, respectively, compared to the case of Fenton reaction only.

• Korean Journal of Microbiology
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• v.38 no.2
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• pp.144-151
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• 2002

For elucidating the relationship between the biosynthetic pathways for polyhydroxyslkanoates (PHAs) and 5-aminolevulinic acid (ALA), culture conditions for the production of these two biomaterials by Rhodopseudomonas sp. KCTC 1437 were investigated. Of the carbon substrates tested, acetic acid was the best carbon source for cell growth and PHA biosynthesis. When succinic acid was added as a co-substrate into culture medium, cell growth and PHA production were greatly increased up to 2.5 g/ι and 73% of dry cell weight, respectively. The PHA obtained from the carbon substrates tested was homopolyester of 3-hydroxybutyrate, while valeric acid was only effective for the production of copolyester consisting of 3-hydroxybutyrate and 3-hydroxyvalerate. Anaerobic light culture condition was better for PHA production and cell growth than anaerobic dark or aerobic dark culture condition. The organism was capable of synthesizing ALA when glycine and succinic acid were added to the culture medium. ALA was produced to ca.400 mg/ι when levulinic acid, soccinic acid, and glycine were repeatedly added with a reductant (sodim thioglycolate). However, the presence of glycine, levulinic acid and sodium glycolate inhibited the cell growth and the conversion of carbon substrates to PHA. From these results it is apparent that the production yields of PHA and ALA could not be increased simultaneously because the optimal conditions for the production of PHA and ALA are opposed to each other.

• 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.

• KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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• v.30 no.1
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• pp.1-14
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• 2024

As awareness of environmental issues continues to grow, there is an escalating demand for recycling and repurposing byproducts of agricultural and food production processes and their conversion to high-value products. Coffee is the most widely consumed beverage globally; during coffee beverage processing and consumption, byproducts such as coffee silverskin (CS), spent coffee grounds (SCGs), and oil are generated. Despite containing beneficial materials such as cellulose, hemicellulose, lignin, lipids, and bioactive substances, these byproducts are typically discarded in landfills or incinerated. The utilization of CS, SCGs, and oil in the development of packaging materials holds significant potentials toward the realization of a sustainable society. To this end, considerable research efforts have been dedicated to the development of high-value materials derived from coffee byproducts, including functional fillers, polymer composites, and biodegradable polymers. Notably, CS and SCGs have been employed as functional fillers in polymer composites. Additionally, lipids extracted from SCGs have been used as plasticizers for polymers and cultured with microorganisms to produce biodegradable polymers. This review focuses on the research and development of polymer/CS and polymer/SCG composites as well as cellulose extraction and utilization from CS and SCGs and its applications, oil extraction from SCGs, and cultivation with microorganisms using extracted oil for polyhydroxyalkanoates(PHA) production.