• Membrane Journal
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• v.33 no.6
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• pp.390-397
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• 2023

Conventional extraction methods for polyhydroxybutyrate (PHB), a sustainable alternative to petroleum-based plastics, cause a decrease in molecular weight and a change in properties. In this work, we developed a method to extract PHB accumulated in microorganisms by physical disruption through filtration using a spiked carbon nanotube (CNT) membrane with functionalized CNT. In addition, filtration of the PHB-containing microbial solution was performed to confirm PHB extraction, which was found to be 4% more efficient than chloroform, the most used extraction method. These results indicate that the spiked CNT membrane has potential in the bioplastics recovery process.

• KSBB Journal
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• v.13 no.1
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• pp.96-100
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• 1998

Two promoters (trc and P$\rho$) were inserted in PHA operon derived from Alcaligenes eutrophus to obtain high chain molecules of polyhydroxybutyrate (PHB) in recombinant Escherichia coli. Newly designed PHA operon was used to control the gene expressions of hydroxybutyric CoA and PHA polymerization, separately. Plasmids containing new synthetic operon was transformed into E. coli DH5$\alpha$ and analyzed for PHB production. Without induction of the PHA biosynthetic operon, PHA synthase which has low activity might supply low concentration of initiator during the polymerization reaction, resulting very high molecular weight of polymer. An increase of PHB average molecular weight was observed with decreased IPTG (isopropyl $\beta$ -Dithiogalactosidase) concentration. When no IPTG was added to the culture of E. coli DH5$\alpha$ /$\rho$ SJS1 which contained two promoters in PHA operon, high chain polymer having an average molecular weight of $2.5{\times}10^7$ was achieved. Analysis of the enzyme activities of PHA biosynthetic enzymes suggests that PHA synthase, the enzyme responsible for polymerizing 3-hydroxybutyric CoA, controls the molecular weight of PHB produced in vivo.

• Journal of Plant Biotechnology
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• v.49 no.3
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• pp.257-270
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• 2022

The aim of this work was to prepare a new biodegradable polyhydroxybutyrate (PHB) submicron fiber mat loaded with hypericin-rich Hypericum perforatum raw extract by centrifugal spinning technology, an alternative approach to the traditional method of electrospinning to fabricate nanofibers or microfibers from solutions at high speed and low cost. Hypericins in methanol/acetone extract of H. perforatum were determined by UHPLC-MS/MS and HPLC/PDA. Submicron fiber mats composed of pure PHB or PHB enriched with H. perforatum extract were prepared using a pilot plant demonstrator for the centrifugal spinning technology and characterized by SEM. Singlet oxygen production was quantified by the 1,3-diphenylisobenzofuran (DPIBF) method in hexane. The results proved a significant production of singlet oxygen by the prepared submicron fiber mat. We also found a significant antibacterial activity against the bacterial strain Escherichia coli CCM 5417 by a method in accordance with JIS Z 2801/ISO 22196 standards. The H. perforatum extract-enriched PHB submicron fiber mats showed potential for the development of self-cleaning and antimicrobial air filters.

• Journal of Environmental Science International
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• v.33 no.2
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• pp.121-129
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• 2024

The study investigated the effect of thermo-alkaline pretreatment on the solubilization of polyhydroxybutyrate (PHB) and its potential to enhance of thermophilic anaerobic digestion, focusing on biochemical methane potential (BMP) and methane production rate, using two different particle sizes of PHB (1500 ㎛ and 400 ㎛). Thermo-alkaline pretreatment tests were conducted at 90 ℃ for 24 hours with varying NaOH dosages from 0-80% (w/w). BMP tests with untreated PHB exhibited methane production ranging from 150.4~225.4 mL CH₄/g COD and 21.5~24.2 mL CH₄/g VSS/d, indicating higher methane production for smaller particle sizes of PHB, 400 ㎛. Thermo-alkaline pretreatment tests achieved a 95.3% PHB solubilization efficiency when 400 ㎛ PHB particles were treated with 80% NaOH dosage at 90 ℃ for 24 hours. BMP tests with pretreated PHB showed substantial improvement in thermophilic anaerobic digestion, with an increase of up to 112% in BMP and up to 168% in methane production rate. The results suggest that a combined pretreatment process, including physical (400 ㎛ PHB particles) and thermo-alkaline (90 ℃, 40-80% NaOH dosage, and 24 hours reaction time), is required for high-rate thermophilic anaerobic digestion of PHB with enhanced methane production.

• Proceedings of the Microbiological Society of Korea Conference
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• 1998.04a
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• pp.47.1-47.1
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• 1998

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.199-199
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• 2005

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.198-198
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• 2005

• Journal of Microbiology and Biotechnology
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• v.29 no.5
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• pp.776-784
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• 2019

Polyhydroxybutyrate (PHB), the most well-known polyhydroxyalkanoate, is a bio-based, biodegradable polymer that has the potential to replace petroleum-based plastics. Lignocellulose hydrolysate, a non-edible resource, is a promising substrate for the sustainable, fermentative production of PHB. However, its application is limited by the generation of inhibitors during the pretreatment processes. In this study, we investigated the feasibility of PHB production in E. coli in the presence of inhibitors found in lignocellulose hydrolysates. Our results show that the introduction of PHB synthetic genes (bktB, phaB, and phaC from Ralstonia eutropha H16) improved cell growth in the presence of the inhibitors such as furfural, 4-hydroxybenzaldehyde, and vanillin, suggesting that PHB synthetic genes confer resistance to these inhibitors. In addition, increased PHB production was observed in the presence of furfural as opposed to the absence of furfural, suggesting that this compound could be used to stimulate PHB production. Our findings indicate that PHB production using lignocellulose hydrolysates in recombinant E. coli could be an innovative strategy for cost-effective PHB production, and PHB could be a good target product from lignocellulose hydrolysates, especially glucose.

• Journal of Environmental Science International
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• v.31 no.7
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• pp.609-616
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• 2022

In this study, the effect of different reaction times for thermal-alkaline pretreatment on the solubilization and biogasification of polyhydroxybutyrate (PHB) were evaluated. Thermal-alkaline pretreatment tests were performed at 73 ℃ and pH 13 at 0-120 h reaction times. The mesophilic anaerobic batch tests were performed with untreated and pretreated PHB samples. The increase in the pretreatment reaction time results in a 52.8-98.8% increase of the abiotic solubilization efficiency of the PHB samples. The reaction time required to achieve solubilization efficiencies of 50%, 90%, and 95% were 10.5, 52.0, and 89.6 h, respectively. The biogasification of the untreated PHB samples achieved a specific methane production rate of 3.6 mL CH₄/g VSS/d and require 101.3 d for complete biogasification. The thermal-alkaline pretreatment significantly improved specific methane production rate (10.2-16.0 time increase), lag time (shortened by 76-81%), and time for complete biogasification (shortened by 21-83%) for the biogasification of the PHB samples when compared to those of the untreated PHB samples. The improvement was higher as the reaction time of the thermal-alkaline pretreatment increased. The findings of this study could be used as a valuable reference for the optimization of the biogasification process in the treatment of PHB wastes.

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.700-709
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• 2024

Polyhydroxybutyrate (PHB) production from lignocellulosic biomass is economically beneficial. Because lignocellulosic biomass is a mixture rich in glucose and xylose, Escherichia coli, which prefers glucose, needs to overcome glucose repression for efficient biosugar use. To avoid glucose repression, here, we overexpressed a xylose regulator (xylR) in an E. coli strain expressing bktB, phaB, and phaC from Cupriavidus necator and evaluated the effect of xylR on PHB production. XylR overexpression increased xylose consumption from 0% to 46.53% and produced 4.45-fold more PHB than the control strain without xylR in a 1% sugar mixture of glucose and xylose (1:1). When the xylR-overexpressed strain was applied to sugars from lignocellulosic biomass, cell growth and PHB production of the strain showed a 4.7-fold increase from the control strain, yielding 2.58 ± 0.02 g/l PHB and 4.43 ± 0.28 g/l dry cell weight in a 1% hydrolysate mixture. XylR overexpression increased the expression of xylose operon genes by up to 1.7-fold. Moreover, the effect of xylR was substantially different in various E. coli strains. Overall, the results showed the effect of xylR overexpression on PHB production in a non-native PHB producer and the possible application of xylR for xylose utilization in E. coli.