Biopolyester Synthesis and Protein Regulations in &lt;I&gt;Ralstonia eutropha&lt;/I&gt; on Levulinic Acid and Its Derivatives from Biomass Refining

Yu, Jian; Chen, Lilian X. L.; Sato, Shun

doi:10.1166/jbmb.2009.1018

Cited by 18 publications

(6 citation statements)

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“…Because of this, economic projections indicate that LA production costs could fall to as low as $0.04–$0.10/lb depending on the scale of operation [20]–[22]. Several studies have demonstrated the use of LA as a sole carbon source or a co-substrate for cell growth and PHA biosynthesis [23]–[24]. LA can serve as a cheap alternative to conventional fermentation substrates.…”

Section: Introductionmentioning

confidence: 99%

Biosynthesis and Thermal Properties of PHBV Produced from Levulinic Acid by Ralstonia eutropha

et al. 2013

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Levulinic acid (LA) can be cost-effectively produced from a vast array of renewable carbohydrate-containing biomaterials. LA could facilitate the commercialization of the polymer poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and PHBV-based products as carbon substrates. Therefore, this paper focused on the production of PHBV by Ralstonia eutropha with LA for hydroxyvalerate (HV) production, which plays an important role in enhancing the thermal properties of PHBV. Accordingly, the HV content of PHBV varied from 0–40.9% at different concentrations of LA. Stimulation of cell growth and PHBV accumulation were observed when 2–6 g L−1 LA was supplied to the culture. The optimal nitrogen sources were determined to be 0.5 g L−1 ammonium chloride and 2 g L−1 casein peptone. It was determined that the optimal pH for cell growth and PHBV accumulation was 7.0. When the cultivation was performed in large scale (2 L fermenter) with a low DO concentration of 30% and a pH of 7.0, a high maximum dry cell weight of 15.53 g L−1 with a PHBV concentration of 12.61 g L−1 (53.9% HV), up to 81.2% of the dry cell weight, was obtained. The melting point of PHBV found to be decreased as the fraction of HV present in the polymer increased, which resulted in an improvement in the ductility and flexibility of the polymer. The results of this study will improve the understanding of the PHBV accumulation and production by R. eutropha and will be valuable for the industrial production of biosynthesized polymers.

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Section: Introductionmentioning

confidence: 99%

Biosynthesis and Thermal Properties of PHBV Produced from Levulinic Acid by Ralstonia eutropha

et al. 2013

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“…Levulinic acid is a platform chemical that can be derived from carbohydrates and cellulosic biomass via hydrolysis 19, 20. It can be used by bacteria to form short‐chain‐length (scl) PHA copolyesters consisting of 3‐hydroxybutyrate (3HB), 3‐hydroxyvalerate (3HV), and 4‐hydroxyvalerate (4HV), as shown in Scheme 21–23. The terpolyester, P(3HB‐ co ‐3HV‐ co ‐4HV), from sole levulinic acid substrate exhibits higher ductility with reduced melting temperature in comparison with P(3HB) and P(3HB‐ co ‐3HV).…”

Section: Introductionmentioning

confidence: 99%

Miscibility of natural polyhydroxyalkanoate blend with controllable material properties

Tanadchangsaeng

2013

J of Applied Polymer Sci

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Polyhydroxyalkanoate (PHA) copolyesters were synthesized by Cupriavidus necator cells in continuous feeding of cosubstrates. During the PHA accumulation phase, the composition of 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), and 4-hydroxyvalerate (4HV) of the copolyesters changed with time, resulting in a change in their miscibility. The as-produced PHA finally became a miscible blend of copolymers with a broad chemical composition distribution. The good miscibility and low crystallinity of the natural P(3HB-co-3HV-co-4HV) blend lead to a remarkable increase in ductility and elongation at break. It indicates that the material properties of copolyesters can be tailored via feeding control of cosubstrates. It was also found that the fractions of natural PHA blend exhibited distinctive thermal behavior and the overall behavior of the as-produced PHA blend was primarily dependent on a fraction of high 3HB content. The material properties of a PHA blend are therefore not determined by its overall chemical composition but more likely by the combined effect of individual copolyesters or fractions. Moreover, the degree of X-ray crystallinity of random P(3HB-co-3HV-co-4HV) blend declined significantly with the increase of 3HV and 4HV content, in contrast to the high crystallinity of well-known P(3HB-co-3HV) copolyesters.

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“…Nowadays, proteomics could determine the entire protein complement of a cell, tissue, or organism under a specific, defined set of conditions [ 19 ]. A number of previous research have investigated the protein regulations during the C. necator grown on various carbon source exposure such as formic acid [ 20 ], gas mixture of H 2 /O 2 /CO 2 [ 18 ], short chain organic acid [ 21 ], as well as levulinate and glucose [ 22 ] via proteomic examination. In this study, a proteomic examination was employed to elucidate the key proteins regulating the pathway shift when the bacterial cells grown on glycerol started producing glucose for maintaining cell growth and PHA formation.…”

Section: Introductionmentioning

confidence: 99%

Proteomic Examination for Gluconeogenesis Pathway-Shift during Polyhydroxyalkanoate Formation in Cupriavidus necator Grown on Glycerol

Tanadchangsaeng

Roytrakul

2020

Bioengineering

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Because of availability and inexpensiveness, glycerol can be considered as a suitable raw material for polyhydroxyalkanoate (PHA) production with bacterial fermentation. Nevertheless, compared to the production of glucose as a raw precursor, PHA produced from glycerol by Cupriavidus necator was found to produce lower PHA with low bacterial growth rates. According to our study, C. necator was able to synthesize glucose-like intermediates from glycerol via gluconeogenesis. This resulted in a decrease of the cell dry weight and the yield of PHA polymers, especially in the active cell growth phase. It was indicated that glycerol used as a carbon source of the PHA synthesis pathway has glucogenesis-shift, which causes a decrease of the PHA content and productivity. In this research, we investigated the proteins that were closely expressed with the increase of the intracellular PHA and glucose content. For solving the above problem, the proteins inside the bacterial cells were analyzed and compared to the database proteins via mass spectrometry. The proteins were isolated by 1-D SDS-polyacrylamide gel electrophoresis (PAGE) technique and identified by the liquid chromatography mass spectrometry (LC-MS) technique. By using bioinformatics validation, a total number of 1361 proteins were examined and found in the culture bacterial cells. Selective protein expression was correlated with the amount of PHA at each cultivation time and generating glucose by studying the 1361 proteins was elucidated in proteomic information. The results of the cluster of proteins were found to contain 93 proteins using the multiple array viewer (MEV) program with the KMS data analysis model. Protein species with the same expression pattern for PHA and six proteins with similar expression patterns were found to be correlated with generating glucose content. The associations of the two protein groups were then determined through a Stitch program. The protein and chemical associations were analyzed both directly and indirectly through different databases. The proteins of interest were found with research data linked between glycerol and glucose. Five protein types are connecting to glucose and glycerol shift pathway, two of which are glycosyl hydrolase (H16_B1563) and short-chain dehydrogenase (H16_B0687), both of which are enzymes used to break the bonds of complex sugars, possibly related to the partial conversion of glycerol to glucose. The two proteins found in the strains used in the Cupriavidus necator H16 experiment give rise to the break down the bonds of α,α-1,1-glucoside of malto-oligosyltrehalose and short-chain sugar molecules such as mannitol (C6H14O6), respectively. In this research, finding the associated expression proteins which is involved in changing the pathway of gluconeogenesis shift to PHA synthesis will be useful information on genetically modifying microorganisms to produce PHA more efficiently, leading to reduction of the production costs.

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Biopolyester Synthesis and Protein Regulations in <I>Ralstonia eutropha</I> on Levulinic Acid and Its Derivatives from Biomass Refining

Cited by 18 publications

References 0 publications

Biosynthesis and Thermal Properties of PHBV Produced from Levulinic Acid by Ralstonia eutropha

Biosynthesis and Thermal Properties of PHBV Produced from Levulinic Acid by Ralstonia eutropha

Miscibility of natural polyhydroxyalkanoate blend with controllable material properties

Proteomic Examination for Gluconeogenesis Pathway-Shift during Polyhydroxyalkanoate Formation in Cupriavidus necator Grown on Glycerol

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