Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by high-cell-density cultivation ofAeromonas hydrophila

Lee, Seung Hwan; Oh, Dong Hyun; Ahn, Woo Suk; Lee, Young; Choi, Jong-il; Lee, Sang Yup

doi:10.1002/(sici)1097-0290(20000120)67:2<240::aid-bit14>3.0.co;2-f

Cited by 118 publications

(85 citation statements)

References 23 publications

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“…P(HB-co-HHx) is mainly produced from related carbon sources such as fatty acids or plant oils in wild-type Aeromonas, Pseudomonas, as well as engineered Ralstonia, Escherichia, and others (Chen et al 2001;Doi et al 1995;Lee et al 2000;Park et al 2001;Qiu et al 2005). However, long-chain fatty acids or plant oils, when used as carbon sources during fermentation, can cause some undesirable problems, such as severe foaming in the fermentation process and challenge in downstream processing of the removal of residual fatty acids (Qiu et al 2005;Riedel et al 2013).…”

Section: Resultsmentioning

confidence: 99%

Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-co-HHx)) from butyrate using engineered Ralstonia eutropha

Jeon

Brigham

Kim

et al. 2014

Appl Microbiol Biotechnol

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Polyhydroxyalkanoates (PHAs), a promising family of bio-based polymers, are considered to be alternatives to traditional petroleum-based plastics. Copolymers like poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-co-HHx)) have been shown to exhibit favorable physical and mechanical properties, due to decreased crystallinity resulting from the presence of medium-chain-length 3-hydroxyhexanoate (3HHx) monomers. In this study, we produced P(HB-coHHx) using engineered Ralstonia eutropha strains containing deletions of the acetoacetyl-CoA reductase (phaB) genes and replacing the native PHA synthase with phaC2 from Rhodococcus aetherivorans I24 and by using butyrate, a short-chain organic acid, as the carbon source. Although the wild-type R. eutropha did not produce P(HB-co-HHx) when grown on mixed acids or on butyrate as the sole carbon source, we are able to produce polymer containing up to 40 wt% 3HHx monomer with the aforementioned engineered R. eutropha strains using various concentrations of just butyrate as the sole carbon source. This is the first report for the production of P(HB-co-HHx) copolymer in R. eutropha using butyrate.

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Section: Resultsmentioning

confidence: 99%

Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-co-HHx)) from butyrate using engineered Ralstonia eutropha

Jeon

Brigham

Kim

et al. 2014

Appl Microbiol Biotechnol

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“…PHA concentration and monomer composition were determined by gas chromatography (Donam Co., Seoul, Korea) equipped with a fused silica capillary column (SPB-5 film [30 m by 0.32 mm; inner diameter, 0.25 m]; Supelco, Bellefonte, Pa.) using benzoic acid as an internal standard (4). Cell concentration, defined as dry cell weight (DCW) per liter of culture broth, was determined as previously described (6,16). The residual cell concentration was defined as the cell concentration minus PHA concentration.…”

Section: Methodsmentioning

confidence: 99%

Identification and Characterization of a New Enoyl Coenzyme A Hydratase Involved in Biosynthesis of Medium-Chain-Length Polyhydroxyalkanoates in Recombinant Escherichia coli

Park

Lee

2003

J Bacteriol

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The biosynthetic pathway of medium-chain-length (MCL) polyhydroxyalkanoates (PHAs) from fatty acids has been established in fadB mutant Escherichia coli strain by expressing the MCL-PHA synthase gene. However, the enzymes that are responsible for the generation of (R)-3-hydroxyacyl coenzyme A (R3HA-CoAs), the substrates for PHA synthase, have not been thoroughly elucidated. Escherichia coli MaoC, which is homologous to Pseudomonas aeruginosa (R)-specific enoyl-CoA hydratase (PhaJ1), was identified and found to be important for PHA biosynthesis in a fadB mutant E. coli strain. When the MCL-PHA synthase gene was introduced, the fadB maoC double-mutant E. coli WB108, which is a derivative of E. coli W3110, accumulated 43% less amount of MCL-PHA from fatty acid compared with the fadB mutant E. coli WB101. The PHA biosynthetic capacity could be restored by plasmid-based expression of the maoC Ec gene in E. coli WB108. Also, E. coli W3110 possessing fully functional ␤-oxidation pathway could produce MCL-PHA from fatty acid by the coexpression of the maoC Ec gene and the MCL-PHA synthase gene. For the enzymatic analysis, MaoC fused with His 6 -Tag at its C-terminal was expressed in E. coli and purified. Enzymatic analysis of tagged MaoC showed that MaoC has enoyl-CoA hydratase activity toward crotonyl-CoA. These results suggest that MaoC is a new enoyl-CoA hydratase involved in supplying (R)-3-hydroxyacyl-CoA from the ␤-oxidation pathway to PHA biosynthetic pathway in the fadB mutant E. coli strain.Polyhydroxyalkanoates (PHAs) are polyesters of (R)-hydroxyalkanoic acids accumulated in numerous bacteria as an energy and carbon storage material under nutrient limiting condition in the presence of excess carbon source (1,17,20). PHAs have been attracting much attention as they can be used as biodegradable polymers (20) and as the sources of chiral pools for the synthesis of fine chemicals (18). The metabolic pathways for the biosynthesis and degradation of PHAs have been well examined in many bacteria (17,20). For example, in short-chain-length-PHA-producing bacteria such as Ralstonia eutropha and Alcaligenes latus, two acetyl coenzyme A (acetylCoA) moieties derived from various carbon sources are condensed to acetoacetyl-CoA by 3-ketothiolase (PhaA) and sequentially converted to (R)-3-hydroxybutyryl-CoA (R3HB-CoA) by acetoacetyl-CoA reductase (PhaB). Then, R3HB-CoA is added to the growing chain of poly(3-hydroxybutyrate) [P(3HB)] by the short-chain-length PHA synthase (PhaC) (29).In pseudomonads belonging to the rRNA homology group I, the intermediates of fatty acid metabolism including enoylCoA, (S)-3-hydroxyacyl-CoA, 3-ketoacyl-CoA, and 3-hydroxyacyl-acyl carrier protein (ACP) are major precursors for medium-chain-length (MCL) PHAs (17,20,36). The metabolic links between the fatty acid metabolism and PHA biosynthesis are mediated by various enzymes such as enoyl-CoA hydratase (7,8,9,23,34,35), 3-ketoacyl-ACP reductase (22, 27, 33), epimerase (20), and 3-hydroxyacyl-ACP:CoA transacylase (12, 26). The genes encoding the...

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“…The short chain length polyhydroxyalkanoates (scl-PHAs), which consist of C 3 -C 5 atoms, medium chain length polyhydroxyalkanoates (mcl-PHAs) consisting of C 6 -C 14 atoms and long chain length polyhydroxyalkanoates (lcl-PHAs) containing >C 14 monomers, and PHAs containing both scl-and mcl-monomer units have been identified in several bacteria (59)(60)(61)(62)(63)(64). In addition, the random copolymer poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(3HB-3HH) or P(HB-HH) was found to be accumulated in some Aeromonas strains (64). This grouping is due to the substrate specificity of the PHA synthesis that only accepts 3-hydroxyalkanoates (3HAs or HAs) of a certain range of carbon length (26).…”

Section: Chemical Structurementioning

confidence: 99%

Bacterial polyhydroxyalkanoates-eco-friendly next generation plastic: Production, biocompatibility, biodegradation, physical properties and applications

Muhammadi

Shabina

Afzal

et al. 2015

Green Chemistry Letters and Reviews

270

139

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Polyhydroxyalkanoates (PHAs) are intracellular aliphatic polyesters synthesized as energy reserves, in the form of water-insoluble, nano-sized discrete and optically dense granules in cytoplasm by a diverse bacteria and some archae under conditions of limiting nutrients in the presence of excess carbon source. Bacteria synthesize different PHAs from coenzyme A thioesters of respective hydroxyalkanoic acid, and degrade intracellularly for reuse and extracellularly in natural environments by other microorganisms. In vivo, PHAs exist as amorphous mobile liquid and water-insoluble inclusions but in vitro, exhibit material and mechanical properties, ranging from stiff and brittle crystalline to elastomeric and molding, similar to petrochemical thermoplastics. Further, they are hydrophobic, isotactic, biocompatible and exhibit piezoelectric properties. But as commodity plastics their applications are limited by high production cost, low yield, in vivo degradation, complexity of technology and difficulty of extraction. Therefore, to replace the conventional plastic with PHAs, it is prerequisite to standardize the PHA production systems.

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Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by high-cell-density cultivation ofAeromonas hydrophila

Cited by 118 publications

References 23 publications

Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-co-HHx)) from butyrate using engineered Ralstonia eutropha

Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-co-HHx)) from butyrate using engineered Ralstonia eutropha

Identification and Characterization of a New Enoyl Coenzyme A Hydratase Involved in Biosynthesis of Medium-Chain-Length Polyhydroxyalkanoates in Recombinant Escherichia coli

Bacterial polyhydroxyalkanoates-eco-friendly next generation plastic: Production, biocompatibility, biodegradation, physical properties and applications

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