• KSBB Journal
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• 제25권3호
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• pp.239-243
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• 2010

식물성 오일 또는 glycerol로부터 Ralstonia eutropha 여섯 균주의 polyhydroxyalkanoates (PHA) 합성 특성을 조사하였다. 탄소원으로 soybean oil, olive oil, 또는 glycerol 만을 공급시 poly(3-hydroxybutyrate) homopolymer가 생성되었으며, $\gamma$-butyrolactone 또는 pentanoic acid를 함께 공급시 poly(3-hydroxybutyrate-co-4-hydroxybutyrate) 또는 poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer가 각각 합성되었다. 최종 균체농도 및 PHA 함량면에서 최적인 균주는 R. eutropha KCTC 2662로 결정되었으며, 최적 기질인 soybean oil 20 g/L로부터 72 h에 균체농도 1.7~9.2 g/L, PHA 함량 70~92 wt%를 얻을 수 있었다.

• KSBB Journal
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• 제29권4호
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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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• 제20권3호
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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.

• 한국미생물·생명공학회지
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• 제32권2호
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• pp.123-127
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• 2004

토양으로부터 oleic acid 를 탄소원으로 이용하여 MCL-PHA의 생산능이 우수한 균주를 분리하여 Pseudomonas sp. MBEL21이라 명명하였다. Pseudomonas sp. MBEL21의 PHA함량을 증가시키기 위하여 여러 가지 필수 영양분의 제한 조건에서 배양한 결과, 인산이 제한된 조건에서 가장 높은 함량의 PHA를 축적하였다. 고농도의 MCL-PHA생산을 위한 Pseudomonas sp. MBEL21의 유가식 배양 전략을 개발하여 28.1 wt％의 함량으로 23 g/L. MCL-PHA를 생산하였다. 또한, 분리된 Pseudomonas sp. MBEL21을 olive oil을 탄소원으로 이용하여 배양한 결과 9.3 wt％의 PHA가 축적되어졌으며, MCL-hydroxyalkanoate와 함께 3-hyoxy-butyrate 의 단량체들로 이루어진 PHA가 확인되어졌다. 이러한 결과들은 분리된 신규 Pseudomonas sp. MBEL21이 저가의 olive oil로부터 생분해성 탄성체의 응용분야를 갖는 MCL-PHA를 효율적으로 생산할 수 있다는 것을 보여준다.

• Journal of Microbiology
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• 제45권2호
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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.

• 한국미생물·생명공학회지
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• 제51권3호
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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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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.207-210
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• 2000

P. oleovorans의 유가식 배양에서 탄소원으로 octanoic acid, 질소원으로 $NH_4NO_3$를 이용한 혼합기질을 배양액의 pH 변화에 따라 공급하는 pH-stat 기질공급전략을 개발하였다. 공급기질의 탄소원/질소원 비 (C/N 비)를 변화시킴으로써 최종 균체농도, PHA 농도, PHA 함량 등을 변화시킬 수 있었으며, 최대 균체농도는 C/N 비가 10 (g octanoic acid/g $NH_4NO_3$)일 때 65 g/L, 최대 PHA 농도는 C/N 비가 20일 때 41 g/L, 최대 PHA 함량은 C/N 비가 20일 때 75%였으며 최대 PHA 생산성은 C/N 비가 10일 때 1.03 g/L/h였다.

• Journal of Microbiology and Biotechnology
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• 제31권2호
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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.

• 생명과학회지
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• 제33권8호
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• pp.680-691
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• 2023

오일 피크, 기후변화 그리고 미세 플라스틱과 관련된 석유합성 플라스틱의 생산과 사용은, 지속 가능한 인류 생활을 위협하는 범세계적 이슈로 대두되고 있다. 이러한 문제를 해결하기 위해 다양한 생분해성 친환경 바이오플라스틱 소재가 대안이 되고 있으며, 그중 주목받고 있는 소재 중 하나는 polyhydroxyalkanoates (PHA) 생분해성 바이오플라스틱이다. 본 총설은 PHA 생산공정에 이용될 수 있는 해양 미생물의 산업적 활용 이점과 현재까지 밝혀진 해양미생물의 생산성을 비교하고, 이들이 산업에서 활용되기 위해 필요한 연구개발 현황에 대해 조사하였다. 조사 결과 해양미생물로부터 생산된 PHA는 석유합성 플라스틱이 보유한 다양한 물리적 특성을 띄는 중합체 소재로의 대체 가능성과, 배지 제조 시 해수를 사용할 수 있는 장점, 특별한 멸균 과정이 필요치 않은 장점, 그리고 분리 정제 과정 등에서 비용 절감의 장점을 제공할 수 있음을 확인하였다. 하지만 해양미생물의 PHA 생산성은 육상에서 분리된 상용화 균주에 비해 다소 떨어지는 효율을 보이는 것을 확인하였다. 이러한 문제점을 해결하기 위해 최근 선진 omics 기반 합성 미생물학 기반 기술을 활용하여, 고밀도 연속배양기술 개발, PHA 효율 증진 및 다양한 시장 요구에 필요한 생분해성 플라스틱 소재 개발이 미래 플라스틱 대체 소재 개발에 필요한 연구임을 확인하였다.

• KSBB Journal
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• 제19권4호
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• pp.331-334
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• 2004

재조합 E. coli를 이용한 MCL-PHA의 생산에서 fatty acid pathway로부터 PHA 생합성 전구체 물질들이 만들어진다는 사실과 함께 이에 관여하는 enzymes이 밝혀지고 있다. 본 논문에서는 protein homology search로부터 탐색된 paaG와 ydbU genes의 PHA 생합성에서의 역할을 확인하기 위하여 paaG와 ydbU gene이 각각 knock-out된 mutant E. coli strains 를 제작하였다. 제작된 mutant E. coli들은 모균주들보다 낮은 PHA 농도와 함량을 가졌으며, 이러한 결과들로부터 paaG와 ydbU는 fatty acid pathway에서 PHA synthesis의 전구체 물질들을 공급한다는 사실을 확인하였다. 또한, 새로운 FadB homologous enzyme YgfG를 탐색하였으며, ygfG gene이 overexpression된 균주와 ygfG mutant를 제작하여 PHA 합성을 실험한 결과 ygfG도 paaG와 ydbU와 유사한 역할을 한다는 사실을 밝혔다. 이러한 연구결과들은 E. coli에서의 MCL-PHA 단량체들의 합성 경로를 확인하여 효과적인 PHA 생산 균주를 제작할 수 있게 할 것이다.