• 제목/요약/키워드: medium-chain-length poly (3-hydroxyalkanoates)

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Pseudomonas sp. EML8 균주를 이용한 폐식용류로부터 medium-chain-length poly(3-hydroxyalkanoates) 생합성 (Production of Medium-chain-length Poly (3-hydroxyalkanoates) by Pseudomonas sp. EML8 from Waste Frying Oil)

  • 김태경;김종식;정정욱
    • 생명과학회지
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    • 제31권1호
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    • pp.90-99
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    • 2021
  • 본 연구에서는 poly (3-hydroxyalkanoate) (PHA)의 생산 비용을 줄이기 위해, 토양에서 분리된 균주 Pseudomonas sp. EML8을 이용하여 폐 튀김유(waste frying oil, WFO)를 단일 탄소원으로 하여 균주의 최적 생장 및 PHA의 생합성 조건을 확립하였다. WFO를 단일 탄소원으로 이용하여 Pseudomonas sp. EML8에 의해 생합성된 PHA를 gas chromatography (GC)와 GC mass spectrometry로 분석한 결과, 7.28 mol% 3-hydrxoyhexanoate, 39.04 mol% 3-hydroxyoctanoate, 37.11 mol% 3-hydroxydecanoate 및 16.58 mol% 3-hydoryxdodecanoate의 단량체로 이루어진 medium-chain-length PHA (mcl-PHAWFO)라는 것을 확인하였다. 플라스크로 배양한 결과, Pseudomonas sp. EML8의 최대 건조세포중량(dry cell weight, DCW) 및 mcl-PHAWFO의 최대 생산수율(g/l)은 WFO 20 g/l, (NH4)2SO4 0.5 g/l, pH 7 및 25℃의 조건에서 확인되었다. 이 결과를 바탕으로 3 l 발효기를 이용하여 48시간 배양한 후에 가장 높은 DCW, mcl-PHAWFO의 함량 및 mcl-PHAWFO의 생산수율(3.0 g/l, 62 wt% 및 1.9 g/l)을 얻었다. 대조군인 신선한 튀김유(fresh frying oil, FFO) 20 g/l를 탄소원으로 사용하여 이와 유사한 DCW, mcl-PHAFFO의 함량 및 mcl-PHAFFO의 생산수율(2.7 g/l, 62 wt%, 1.6 g/l)을 확인했다. 겔 투과 크로마토그래피 분석을 통해 mcl-PHAWFO 및 mcl-PHAFFO의 평균 분자량이 165-175 kDa인 것을 확인했으며, 열중량을 분석한 결과, mcl-PHAWFO 및 mcl-PHAFFO는 각각 260 및 274.7℃의 분해온도값을 보여주었다. 결론적으로 본 연구에서는 Pseudomonas sp. ML8과 WFO는 mcl-PHA의 생산을 위한 새로운 균주와 탄소원으로서의 사용 가능성을 확인하였다.

Isolation of a Pseudomonas sp. Strain Exhibiting Unusual Behavior of Poly(3-hydroxyalkanoates) Biosynthesis and Characterization of Synthesized Polyesters

  • Chung, Chung-Wook;Kim, Yoon-Seok;Kim, Young-Baek;Bae, Kyung-Sook;Rhee, Young-Ha
    • Journal of Microbiology and Biotechnology
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    • 제9권6호
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    • pp.847-853
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    • 1999
  • A Pseudomonas sp. strain that is capable of utilizing dicarboxylic acids as a sole carbon source was isolated from activated sludge by using the enrichment culture technique. This organism accumulated polyhydroxyalkanoates (PHAs) with an unusual pattern of monomer units that depends on the carbon sources used. Polyhydroxybutyrate (PHB) homopolyester was synthesized from glucose or small $C_{-even}$ alkanoic acids, such as butyric acid and hexanoic acid. Accumulation of PHB homopolyester was also observed in the cells grown on $C_{-odd}$ dicarboxylic acids, such as heptanedioic acid and nonanedioic acid as the sole carbon sources. In contrast, a copolyester consisting of 6 mol% 3-hydroxybutyrate (3HB) and 94 mol% 3-hydroxyvalerate (3HV) was produced with a PHA content of as much as 36% of the cellular dry matter. This strain produced PHAs consisting both of the short-chain-length (SCL) and the medium-chain-length (MCL) 3-hydroxyacid units when heptanoic acid to undecanoic acid were fed as the sole carbon sources. Most interestingly, polyester consisting of significant amount of relevant fractions, 3HB, 3HV, and 3-hydroxyheptanoate (3HHp), was accumulated from heptanoic acid. According to solvent fractionation experiments, the polymer produced from heptanoic acid was a blend of poly(3HHp) and of a copolyester of 3HB, 3HV, and 3HHp units. The hexane soluble fractions contained only 3HHp units while the hexane-insoluble fractions contained 3HB and 3HV units with a small amount of 3HHp unit. The copolyester was an elastomer with unusual mechanical properties. The maximum elongation ratio of the copolyester was 460% with an ultimate strength of 10 MPa, which was very different from those of poly(3HB-co-3HV) copolyesters having similar compositions produced from other microorganisms.

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Biosynthesis of medium-chain-length poly(3-hydroxyalkanoates) by metabolically engineered Escherichia coli strains

  • 박시재;이상엽
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2001년도 추계학술발표대회
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    • pp.735-738
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    • 2001
  • Metabolically engineered Escherichia coli strains harboring a plasmid containing a novel artificial polyhydroxyalkanoate (PHA) operon consisting of the Aeromonas PHA biosynthesis related genes and Ralstonia eutropha reductase gene were developed for the production of poly(3-hydroxybutyrate-co-hydroxyhexanoate) [P(3HB-co-3HHx)] from dodecanoic acid. By applying stepwise reduction of dissolved oxygen concentration (DOC) during the fermentation, the final dry cell weight, PHA concentration, and PHA content of 79 g/L, 21.5 g/L, and 27.2 wt%, respectively, were obtained in 40.8 h, which resulted in the PHA productivity of 0.53 g/L/h. The 3HHx fraction slowly increased during the fed-batch culture to reach a final value of 10.8 mol%. The 3HHx fraction in the copolymer could be increased by three fold when the Aeromonas hydrophila orfl gene was co-expressed with the PHA biosynthesis genes.

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High Cell Density Cultivation of Pseudomonas oleovorans for the Production of Poly(3-Hydroxyalkanoates)

  • Lee, Sang-Yup
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제1권1호
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    • pp.51-53
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    • 1996
  • Fed-batch culture of Pseudomonas oleovorans was carried out for the production of medium-chain-length polyhydroxyalkanoates (MCL-PHAs) using octanoate as a carbon source. Octanoate and the salt solution containing ammounium sulfate and magnesium sulfate were intermittently fed in the course of fermentation. Cell mass and PHA concentrations of 42.8 and 16.8g/L, respectively, could be obtained in 40 h. The PHA content and the PHA productivity were 39.2% and 0.42 g PHA/L-h, respectively. The yields of cell mass and PHA were 0.71 g dry cell mass/g octanoate and 0.28g PHA/g octanoate, respectively. Therefore, octanoate can be used for the production of MCL-PHAs to a high concentration with high productivity.

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