• 한국미생물·생명공학회지
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• 제28권2호
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• pp.71-79
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• 2000

Two typess of copolyesters, poly(3-hydroxybutyric acid-co-4-hydroxy-butyric acid)[P(3HB-co-4HB] and poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid)[P(3HB-co-3HV)], with various monomer ratios and different degree of microstructural heterogeneity were synthesized from Ralstonia eutropha H16 and Hydrogenophaga pseudoflava by using ${\gamma}$-butyrolactone and ${\gamma}$-valerolactone, respectively. The two bacteria showed a large difference in the utilization of ${\gamma}$-butyrolactone for cell growth and PHA synthesis. H. pseudoflava synthesized P(3HB-co-4HB) copolyesters with a wide range of 4HB content from 13 to 96 mol% depending on culture conditions, whiel R. eutropha H16 was able to synthesize the copolyesters containing less than 20 mol% of 4HB. An increase in the 4HB content in the P(3HB-co-4HB) copolyesters synthesized by H. pseud-oflava induced an lowering of their melting temperatures as well as their enthalpies of fusion. The increase in the 4HB content, however, increased the rate of degradation by an extracellular P(3HB) depolymerase. NMR spectros-copy and differential scanning calorimetry showed that the P(3HB-co-4HB) copolyesters from H. pseudoflava were generally microstructurally heterogeneous. The P(3HB-co-4HB) copolyesters) synthesized by R. eutropha H16 were rather random copolymers showing less microstructural heterogeneity than those synthesized by H. pseudoflava. The NMR D value analysis suggested that the monomer distribution of the P(3HB-co-3HV) copolymers from the two bacteria were relatively random.

• 대한환경공학회지
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• 제28권11호
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• pp.1222-1227
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• 2006

중금속으로 오염된 폐 광산에서 중금속(카드뮴, 구리)에 내성을 가지는 총 9개의 균주를 순수 분리한 후, 스크린 테스트를 거쳐 고효율의 중금속 제거 균주 JH1을 선별하였다. JH1 균주는 형태학적, 생리 생화학적 특징, 지방산 분석 그리고 16S rDNA 분석결과 Ralstonia eutropha JH1으로 동정되었다. 카드뮴으로 오염된 토양을 구연산(10 mM, pH 6.0)으로 세척 한 후, 그 세척수(카드뮴 110 mg/L)에서 균 농도(0.5, 1.0, 2.0, 4.0 g/L)에 따른 카드뮴 제거율을 확인한 결과, 5일 동안 각각 49.9, 84.4, 89.7, 89.9% 제거되었다. 또한 카드뮴(110 mg/L)과 구리(100 mg/L)를 함유하고 있는 구연산-세척수에서 Ralstonia eutropha JH1을 1.0 g/L로 접종했을 때, 6일 동안 카드뮴과 구리가 100% 제거 되었다. 저농도(10, 30, 60 mg/L)의 카드뮴과 구리를 각각 함유한 세척수 내에 Ralstonia eutropha JH1을 1.0 g/L로 접종하고 시간에 따른 제거율을 확인한 결과, 카드뮴과 구리 모두 10, 30, 60 mg/L에서 각각 12, 18, 48시간에서 100%제거되었다.

• Journal of Microbiology and Biotechnology
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• 제29권10호
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• pp.1656-1664
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• 2019

Isoprene has the potential to replace some petroleum-based chemicals and can be produced through biological systems using renewable carbon sources. Ralstonia eutropha can produce value-added compounds, including intracellular polyhydroxyalkanoate (PHA) through fatty acid and lipid metabolism. In the present study, we engineered strains of R. eutropha H16 and examined the strains for isoprene production. We optimized codons of all the genes involved in isoprene synthesis by the mevalonate pathway and manipulated the promoter regions using pLac and pJ5 elements. Our results showed that isoprene productivity was higher using the J5 promoter ($1.9{\pm}0.24{\mu}g/l$) than when using the lac promoter ($1.5{\pm}0.2{\mu}g/l$). Additionally, the use of three J5 promoters was more efficient ($3.8{\pm}0.18{\mu}g/l$) for isoprene production than a one-promoter system, and could be scaled up to a 5-L batch-cultivation from a T-flask culture. Although the isoprene yield obtained in our study was insufficient to meet industrial demands, our study, for the first time, shows that R. eutropha can be modified for efficient isoprene production and lays the foundation for further optimization of the fermentation process.

• Journal of Microbiology and Biotechnology
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• 제18권8호
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• pp.1408-1415
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• 2008

To produce polyhydroxyalkanoate (PHA) from inexpensive substrates by bacteria, vegetable-oil-degrading bacteria were isolated from a rice field using enrichment cultivation. The isolated Pseudomonas sp. strain DR2 showed clear orange or red spots of accumulated PHA granules when grown on phosphate and nitrogen limited medium containing vegetable oil as the sole carbon source and stained with Nile blue A. Up to 37.34% (w/w) of intracellular PHA was produced from corn oil, which consisted of three major 3-hydroxyalkanoates; octanoic (C8:0, 37.75% of the total 3-hydroxyalkanoate content of PHA), decanoic (C10:0, 36.74%), and dodecanoic (C12:0, 11.36%). Pseudomonas sp. strain DR2 accumulated up to 23.52% (w/w) of $PHA_{MCL}$ from waste vegetable oil. The proportion of 3-hydroxyalkanoate of the waste vegetable-oil-derived PHA [hexanoic (5.86%), octanoic (45.67%), decanoic (34.88%), tetradecanoic (8.35%), and hexadecanoic (5.24%)] showed a composition ratio different from that of the corn-oil-derived PHA. Strain DR2 used three major fatty acids in the same ratio, and linoleic acid was the major source of PHA production. Interestingly, the production of PHA in Pseudomonas sp. strain DR2 could not occur in either acetate- or butyrate-amended media. Pseudomonas sp. strain DR2 accumulated a greater amount of PHA than other well-studied strains (Chromobacterium violaceum and Ralstonia eutropha H16) when grown on vegetable oil. The data showed that Pseudomonas sp. strain DR2 was capable of producing PHA from waste vegetable oil.

• Journal of Microbiology and Biotechnology
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• 제29권5호
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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.