Analysis of Poly-β-Hydroxybutyrate in
            <i>Rhizobium japonicum</i>
            Bacteroids by Ion-Exclusion High-Pressure Liquid Chromatography and UV Detection

Karr, Dale B.; Waters, James K.; Emerich, David W.

doi:10.1128/aem.46.6.1339-1344.1983

Cited by 283 publications

(116 citation statements)

References 16 publications

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“…Glucose and extracellular metabolites including ethanol, glycerol, and acetate were quantified in the culture supernatant using an Ultimate 3000 HPLC (Dionex, Sunnyvale, CA) equipped with an Aminex HPX 87H ion exclusion column (300 mm Â 7.8 mm, Bio-Rad Laboratories, Hercules, CA) which was operated at 458C and a flow rate of 0.6 mL/min of 5 mM H 2 SO 4 using a refractive index detector and UV detector for analysis of sugars and organic acids, respectively. PHB was analyzed as described previously (Karr et al, 1983;Tyo et al, 2006); 10-20 mg of dried cells were weighed and boiled in 1 mL of concentrated sulfuric acid for 60 min and then diluted with 4 mL of 14 mM H 2 SO 4 . Samples were centrifuged (15 min, 16,000g) to remove cell debris, and the supernatant was analyzed using an Ultimate 3000 HPLC (Dionex) equipped with an Aminex HPX-87H ion exclusion column (300 Â 7.8 mm; Bio-Rad Laboratories) and UV detector.…”

Section: Metabolite Analysismentioning

confidence: 99%

Improved polyhydroxybutyrate production by Saccharomyces cerevisiae through the use of the phosphoketolase pathway

Kocharin

Siewers

Nielsen

2013

Biotech & Bioengineering

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The metabolic pathways of the central carbon metabolism in Saccharomyces cerevisiae are well studied and consequently S. cerevisiae has been widely evaluated as a cell factory for many industrial biological products. In this study, we investigated the effect of engineering the supply of precursor, acetyl-CoA, and cofactor, NADPH, on the biosynthesis of the bacterial biopolymer polyhydroxybutyrate (PHB), in S. cerevisiae. Supply of acetyl-CoA was engineered by over-expression of genes from the ethanol degradation pathway or by heterologous expression of the phophoketolase pathway from Aspergillus nidulans. Both strategies improved the production of PHB. Integration of gapN encoding NADP(+) -dependent glyceraldehyde-3-phosphate dehydrogenase from Streptococcus mutans into the genome enabled an increased supply of NADPH resulting in a decrease in glycerol production and increased production of PHB. The strategy that resulted in the highest PHB production after 100 h was with a strain harboring the phosphoketolase pathway to supply acetyl-CoA without the need of increased NADPH production by gapN integration. The results from this study imply that during the exponential growth on glucose, the biosynthesis of PHB in S. cerevisiae is likely to be limited by the supply of NADPH whereas supply of acetyl-CoA as precursor plays a more important role in the improvement of PHB production during growth on ethanol.

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Section: Metabolite Analysismentioning

confidence: 99%

Improved polyhydroxybutyrate production by Saccharomyces cerevisiae through the use of the phosphoketolase pathway

Kocharin

Siewers

Nielsen

2013

Biotech & Bioengineering

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“…For quantitative chemical determinations, the dried sample may be hydrolyzed in concentrated sulfuric acid to convert total PHB to crotonic acid [8]. The crotonic acid can then be isolated and quantified [6][7][8][9] with a sensitivity limit of 10 ng PHB. In using this procedure, it is necessary to rule out or evaluate the contribution of the monomer, /3-hydroxybutyrate.…”

Section: Analysis Of Cphbmentioning

confidence: 99%

Biological complexes of poly-β-hydroxybutyrate

Reusch¹

1992

FEMS Microbiology Letters

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“…Presence of PHAs in the extracted sample was confirmed by Fourier transform infrared (FT-IR) spectroscopic analysis. PHAs quantification was carried out by HPLC (Shimadzu LC10 A) according to procedure outlined by Karr et al [26]. Standard curve was plotted using pure co-polymer poly-3(hydroxybutyrate-co-hydroxyvalerate) purchased from Aldrich company.…”

Section: Biopolymer Extractionmentioning

confidence: 99%

Bacterial synthesis of polyhydroxyalkanoates using dark fermentation effluents: Comparison between pure and enriched mixed cultures

Reddy

Amulya

Mohan

2015

Engineering in Life Sciences

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Polyhydroxyalkanoates (PHAs) are naturally occurring biopolymeric materials synthesized by a wide group of microorganisms. PHAs are known for their use as bioplastics as they have similar physicochemical properties to those of conventional plastics. In the present scenario, PHAs are produced by mixed bacterial cultures or pure cultures using various types of substrates. The present study aimed at investigating the ability of bacteria isolated from an enriched mixed culture to produce PHAs using both synthetic wastewater and effluents collected from hydrogen‐producing reactor as substrates. PHAs production was performed using both the enriched mixed culture and the isolated strain Serratia ureilytica under nitrogen limitation aiming to compare PHAs accumulation capacity. The use of enriched mixed culture and synthetic wastewater resulted in the highest PHAs accumulation, i.e. 54 ± 3% dry cell weight (DCW). However, when S. ureilytica was used, PHAs accumulation decreased, i.e. 51 ± 2% DCW with synthetic acids, and 31 ± 2% DCW with effluents collected from hydrogen‐producing reactor. Produced polymer contained the co‐polymer with monomer composition of poly‐3(hydroxybutyrate‐co‐hydroxyvalerate). Food waste as substrate contributes in reducing the production cost of both hydrogen as well as PHAs production embedded with waste valorization.

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Analysis of Poly-β-Hydroxybutyrate in Rhizobium japonicum Bacteroids by Ion-Exclusion High-Pressure Liquid Chromatography and UV Detection

Cited by 283 publications

References 16 publications

Improved polyhydroxybutyrate production by Saccharomyces cerevisiae through the use of the phosphoketolase pathway

Improved polyhydroxybutyrate production by Saccharomyces cerevisiae through the use of the phosphoketolase pathway

Biological complexes of poly-β-hydroxybutyrate

Bacterial synthesis of polyhydroxyalkanoates using dark fermentation effluents: Comparison between pure and enriched mixed cultures

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