Possibilities for controlling a PHB accumulation process using various analytical methods

Wendlandt, K.‐D.; Geyer, W.; Mirschel, Gabriele; Hemidi, F.A.-H.

doi:10.1016/j.jbiotec.2005.01.007

Cited by 49 publications

(27 citation statements)

References 19 publications

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“…cultivated on rice, canola, sunflower, soybean, corn and hydrolyzed linseed oils [6,18]. Polymer accumulation, indicated by Nile blue A (Nile blue sulphate, basic blue) is consistent with previous reports and was a good indicator since stained PHA granules are easily identifiable by their characteristic orange fluorescence [16,47]. Neutral lipids do not affect identification of PHA since these lipids are liquids at the staining temperature (55°C) and have no affinity for Nile red (the oxidized form of Nile blue); therefore, cell membranes and other lipid containing cell components do not absorb enough dye to yield a detectable fluorescence at a wavelength of 460 nm [46].…”

Section: Discussionsupporting

confidence: 82%

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Non-Edible Vernonia galamensis Oil and Mixed Bacterial Cultures for the Production of Polyhydroxyalkanoates

Allen¹

2014

Mod Chem Appl

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Since the oil crisis of the 1970s much attempt has been made, albeit with varying degrees of success, to source the ideal substrate and bacteria for the production of PHA. The non-edible, naturally epoxidized seed oil from Vernonia galamensis and mixed cultures consisting of Alcaligenes latus (ATCC 29712), Cupriavidus necator (ATCC 17699), Escherichia coli (DH5α) and Pseudomonas oleovorans (ATCC 29347), were evaluated for PHA production under batch and fed-batch fermentations. PHA production, optimized by the mixed culture of E. coli and C. necator, was 0.4-19% (%wt/wt, cdw) for batch and fed-batch fermentations. Analyses of PHA by Matrix Assisted Laser Desorption IonizationTime of Flight Mass Spectrometry (MALDI-TOF MS) and Gas Chromatography Mass Spectrometry (GC/MS) identified the 3-hydroxybutyrate (3HB) monomeric unit. The PHA ester bond stretching vibration (C=O), was confirmed at absorption 1740.66 cm -1 , using Fourier Transform Infrared Spectroscopy (FTIR). Gel Permeation Chromatography (GPC) indicated peak molecular weights between 3.8×10 3 -1.12×10 6 Da with melting points (T m ), 60-90°C. The data further illustrates that inedible oils could be the ideal carbon source for the production of PHA.

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Section: Discussionsupporting

confidence: 82%

“…Throughout fermentations, 500 µl broth was evaluated for PHA accumulation using Nile blue A and the hypochlorite assay [46][47][48]. Smears were stained with Nile blue A (1%) at 55°C for 10 min and slides washed with water and 8% acetic acid for 1 min then evaluated.…”

Section: Batch/fed-batch Fermentation Extraction and Purification Ofmentioning

confidence: 99%

Non-Edible Vernonia galamensis Oil and Mixed Bacterial Cultures for the Production of Polyhydroxyalkanoates

Allen¹

2014

Mod Chem Appl

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“…At the large scale considered here, operating the bioreactors at the dilution rate of 0.17 /h reported by Wendlandt et al (24,25) may be unrealistic due to mass transfer limitations, so a conservative value of 0.085 /h was assumed. The bacteria will store low levels of PHB in the growth phase (assumed PHA content in biomass of 3 wt%).…”

Section: Bioreactorsmentioning

confidence: 99%

Techno-economic assessment of poly-3-hydroxybutyrate (PHB) production from methane—The case for thermophilic bioprocessing

Levett

Birkett

Davies

et al. 2016

Journal of Environmental Chemical Engineering

121

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, Techno-economic assessment of poly-3-hydroxybutyrate (PHB) production from methane-The case for thermophilic bioprocessing, Journal of Environmental Chemical Engineering http://dx.doi.org/10.1016/j.jece. 2016.07.033 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. AbstractA major obstacle preventing the large scale production of polyhydroxyalkanoates (PHAs) has been the lack of a reliable, low cost, large volume feedstock. The abundance and relatively low price of methane therefore marks it as a substrate of interest. This paper presents a technoeconomic assessment of the production of poly-3-hydroxybutyrate (PHB) from methane.ASPEN Plus was used for process design and simulation. The design and economic evaluation is presented for production of 100,000 t/a PHB through methanotrophic fermentation and acetone-water solvent extraction. Production costs were estimated at $4.1-$6.8/kg PHA, which compares against a median price of $7.5/kg from other studies. Raw material costs are reduced from 30-50% of production for sugar feedstocks, to 22% of production for methane. A feature of the work is the revelation that heat removal from the two-stage bioreactor process contributes 28% of the operating cost. Thermophilic methanotrophs could allow the use of cooling water instead of refrigerant, reducing production costs to $3.2-5.4/kg PHA; it is noted that PHB producing thermophilic methanotrophs are yet to be isolated. Energy consumption for air compression and biomass drying were also identified as significant capital and operating costs and therefore optimisation of bioreactor height and pressure and biomass moisture content should be considered in future research.3

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“…Assays of poly-b-hydroxybutyrate (PHB), dry cell weight and glucose PHB was measured according to the methods of Wendlandt et al (2005). CDW was determined by drying the cells at 105°C to a constant weight.…”

Section: Analysis Of Extracellular Metabolitesmentioning

confidence: 99%

A fundamental dual regulatory role of citrate on the biosyntheses of thuringiensin and poly-β-hydroxybutyrate in Bacillus thuringiensis YBT-032

et al. 2007

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The production of alpha-ketoglutarate, adenine, thuringiensin production rate and thuringiensin yield on glucose consumed increased by 22%, 36%, 40% and 40%, respectively, in presence of 2 g citrate/l. However, citrate decreased pyruvate production, poly-beta-hydroxybutyrate (PHB) production rate and PHB yield by 62%, 31% and 45%, respectively. The activities of pyruvate kinase and glucose-6-phosphate dehydrogenase were 36%-45% lower and 50%-120% higher than those of the control, respectively. The results suggest that citrate regulated the carbon flux to synthesis of adenine present in thuringiensin with a higher efficiency of utilization of glucose by decreasing PHB synthesis.

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Possibilities for controlling a PHB accumulation process using various analytical methods

Cited by 49 publications

References 19 publications

Non-Edible Vernonia galamensis Oil and Mixed Bacterial Cultures for the Production of Polyhydroxyalkanoates

Non-Edible Vernonia galamensis Oil and Mixed Bacterial Cultures for the Production of Polyhydroxyalkanoates

Techno-economic assessment of poly-3-hydroxybutyrate (PHB) production from methane—The case for thermophilic bioprocessing

A fundamental dual regulatory role of citrate on the biosyntheses of thuringiensin and poly-β-hydroxybutyrate in Bacillus thuringiensis YBT-032

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