Cupriavidus necator B-10646 growth and polyhydroxyalkanoates production on different plant oils.

The study addresses the growth of the wild-type strain Cupriavidus necator B-10646 and synthesis of polyhydroxyalkanoates by this strain on media containing plant oils with different compositions of fatty acids: palm, Siberian oilseed, and refined and unrefined sunflower seed oils. The study showed that the best carbon substrate was palm oil. Comparison of fatty acid compositions of the starting oils and unutilized residual substrates showed that C. necator B-10646 cells consumed the fatty acids from palm oil evenly while in experiments with other oils, they utilized polyenoic fatty acids first. Higher production parameters of the culture were obtained by preparation of emulsified oil medium using Tween 80 and sodium cocoyl glutamate as emulsifiers. All polyhydroxyalkanoate specimens were terpolymers that contained 3-hydroxybutyrate as the major component and minor amounts of 3-hydroxyvalerate (0.9-1.9 mol%) and 3-hydroxyhexanoate (0.5-1.1 mol%). Molecular weight of polyhydroxyalkanoate specimens depended on the type of plant oil and emulsifier.

Diclofenac inhibited the biological phosphorus removal: Performance and mechanism.

This work aims to evaluate the effect of new contaminant diclofenac (DCF) in sewage on the performance of Enhanced Biological Phosphorus Removal (EBPR) and its mechanism. The results showed that low-level DCF had no significant effect on EBPR. However, when the concentration of DCF was 2.0 mg/L, the removal efficiencies of chemical oxygen demand (COD), NH4+-N and soluble orthophosphate (SOP) decreased significantly to 71.2 ± 4.2%, 78.6 ± 2.9%, and 64.3 ± 4.2%, respectively. Mechanisms revealed that DCF promoted the ratio of protein to polysaccharide in activated sludge extracellular polymers and inhibited anaerobic phosphorus release and oxic phosphorus uptake. Intracellular polymer analysis showed that when the DCF content was 2.0 mg/L, the maximum content of polyhydroxyalkanoates (PHA) was only 2.5 ± 0.4 mmol-C/g VSS, which was significantly lower than that in the blank. Analysis of key enzyme activities indicated that the presence of DCF reduced the activities of exopolyphosphatase and polyphosphate kinase.

Production of polyhydroxyalkanoate (PHA) by Ralstonia eutropha JMP 134 with volatile fatty acids from palm oil mill effluent as precursors.

The highest volatile fatty acids (VFAs) concentration from palm oil mill effluent (POME) treated by anaerobic fermentation was achieved for a 1-day process when the main acids used were acetic, propionic and butyric acids. Polyhydroxyalkanoate (PHA) production with VFAs from POME as precursors in the fed-batch mode has advantages over batch mode, both in terms of its productivity and 3HV (3-hydroxyvalerate) composition in the produced polymer. With the fed batch, the productivity increased to 343% and contained more 3HV than those of the batch. The structures of the PHA were identified by different methods and they supported each other; the resulting products consisted of functional groups of 3HB (3-hydroxybutyrate) and 3HV.

Analysis of poly-ß-hydroxyalkonates (PHA) during the enhanced biological phosphorus removal process using FTIR spectroscopy.

Enhanced biological phosphorus removal (EBPR) is the main phosphorus removal technique for wastewater treatment. During the anaerobic-aerobic alternative process, the activated sludge experienced the anaerobic storage of polyhydroxy-ß-alkonates (PHA) and aerobic degradation, corresponding the infrared peak intensity of sludge at 1 740 cm(-1) increased in the aerobic phase and declined in the anaerobic phase. Compared with PHA standard, this peak was indentified to attribute the carbonyl of PHA. The overlapping peaks of PHA, protein I and II bands were separated using Gaussian peak fitting method. The infrared peak area ratios of PHA versus protein I had a good relationship with the PHA contents measured by gas chromatography, and the correlation coefficient was 0.873. Thus, the ratio of the peak area of PHA versus protein I can be considered as the indicator of the PHA content in the sludge. The infrared spectra of 1 480-1 780 cm(-1) was selected, normalized and transferred to the absorption data. Combined with the chromatography analysis of PHA content in the sludge sample, a model between the Fourier-transform infrared spectroscopy (ETIR) spectra of the sludge and PHA content was established, which could be used for the prediction of the PHA content in the unknown sample. The PHA content in the sludge sample could be acquired by the infrared spectra of the sludge sample and the established model, and the values fitted well with the results obtained from chromatograph. The results would provide a novel analysis method for the rapid characterization and quantitative determination of the intracellular PHA content in the activated sludge.

Pulsed feeding strategy is more favorable to medium-chain-length polyhydroxyalkanoates production from waste rapeseed oil.

This article presents the results of production and characterization of medium-chain-length polyhydroxyalkanoates (mcl-PHAs) using Pseudomonas sp. Gl01. Studies have been carried out to find suitable feeding strategies for mcl-PHAs production and, for the first time, to investigate in-depth the properties of biopolyesters obtained under controlled conditions with waste rapeseed oil as a substrate. Up to 44% mcl-PHAs of cell dry weight was produced at 41 h of biofermentor culture by employing pulsed feeding of waste rapeseed oil. GC analysis showed a polymer composition with monomer length of C6 to C12 with C8 and C10 as the principal monomers. The monomeric structure of the extracted polyesters did not depend on the cultivation time and the feeding strategy. Molecular weight of the mcl-PHAs was found to be ranging from 57 to 154 kDa. Thermal analyses showed the obtained mcl-polyhydroxyalkanaotes to be semi-crystalline biopolymer with promising thermal stability, having a glass transition temperature of -38 to -50°C.

Temperature influence on biological phosphorus removal induced by aerobic/extended-idle regime.

Previous researches have demonstrated that biological phosphorus removal (BPR) from wastewater could be driven by the aerobic/extended-idle (A/EI) regime. This study further investigated temperature effects on phosphorus removal performance in six A/EI sequencing batch reactors (SBRs) operated at temperatures ranging from 5 to 30 °C. The results showed that phosphorus removal efficiency increased with temperature increasing from 5 to 20 °C but slightly decreased when temperature continually increased to 30 °C. The highest phosphorus removal rate of 97.1 % was obtained at 20 °C. The biomass cultured at 20 °C contained more polyphosphate accumulating organisms (PAO) and less glycogen accumulating organisms (GAO) than that cultured at any other temperatures investigated. The mechanism studies revealed that temperature affected the transformations of glycogen and polyhydroxyalkanoates, and the activities of exopolyphosphatase and polyphosphate kinase activities. In addition, phosphorus removal performances of the A/EI and traditional anaerobic/oxic (A/O) SBRs were compared at 5 and 20 °C, respectively. The results showed the A/EI regime drove better phosphorus removal than the A/O regime at both 5 and 20 °C, and more PAO and less GAO abundances in the biomass might be the principal reason for the higher BPR in the A/EI SBRs as compared with the A/O SBRs.

Production of polyhydroxyalkanoates (PHAs) with canola oil as carbon source.

Wautersia eutropha was able to synthesize medium chain length polyhydroxyalkanoates (PHAs) when canola oil was used as carbon source. W. eutropha was cultivated using fructose and ammonium sulphate as carbon and nitrogen sources, respectively, for growth and inoculum development. The experiments were done in a laboratory scale bioreactor in three stages. Initially, the biomass was adapted in a batch culture. Secondly, a fed-batch was used to increase the cell dry weight and PHA concentration to 4.36 g L(-1) and 0.36 g L(-1), respectively. Finally, after the addition of canola oil as carbon source a final concentration of 18.27 g L(-1) PHA was obtained after 40 h of fermentation. With canola oil as carbon source, the polymer content of the cell dry matter was 90%. The polymer was purified from dried cells and analyzed by FTIR, NMR and DSC using PHB as reference. The polymer produced by W. eutropha from canola oil had four carbon monomers in the structure of the PHA and identified by 1H and 13C NMR analysis as 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), 3-hydroxyoctanoate (3HO), and 3-hydroxydodecanoate (3HDD).

Biological treatment of two-phase olive mill wastewater (TPOMW, alpeorujo): polyhydroxyalkanoates (PHAs) production by Azotobacter strains.

Azotobacter chroococcum H23 (CECT 4435), Azotobacter vinelandii UWD, and Azotobacter vinelandii (ATCC 12837), members of the family Pseudomonadaceae, were used to evaluate their capacity to grow and accumulate polyhydroxyalkanoates (PHAs) using two-phase olive mill wastewater (TPOMW, alpeorujo) diluted at different concentrations as the sole carbon source. The PHAs amounts (g/l) increased clearly when the TPOMW samples were previously digested under anaerobic conditions. The MNR analysis demonstrated that the bacterial strains formed only homopolymers containing beta-hydroxybutyrate, either when grown in diluted TPOMW medium or diluted anaerobically digested TPOMW medium. COD values of the diluted anaerobically digested waste were measured before and after the aerobic PHA-storing phase, and a clear reduction (72%) was recorded after 72 h of incubation. The results obtained in this study suggest the perspectives for using these bacterial strains to produce PHAs from TPOMW, and in parallel, contribute efficiently to the bioremediation of this waste. This fact seems essential if bioplastics are to become competitive products.

PHA(MCL) biosynthesis systems in Pseudomonas aeruginosa and Pseudomonas putida strains show differences on monomer specificities.

The production of PHA from plant oils by Pseudomonas species soil isolated from a sugarcane crop was evaluated. Out of 22 bacterial strains three were able to use efficiently plant oils to grow and to accumulate PHA. Pseudomonas putida and Pseudomonas aeruginosa strains produced PHA presenting differences on monomer composition compatible with variability on monomer specificity of their PHA biosynthesis system. The molar fraction of 3-hydroxydodecanoate detected in the PHA was linearly correlated to the oleic acid supplied. A non-linear relationship between the molar fractions of 3-hydroxy-6-dodecenoate (3HDdDelta(6)) detected in PHA and the linoleic acid supplied was observed, compatible with saturation in the biosynthesis system capability to channel intermediate of beta-oxidation to PHA synthesis. Although P. putida showed a higher 3HDdDelta(6) yield from linoleic acid when compared to P. aeruginosa, in both species it was less than 10% of the maximum theoretical value. These results contribute to the knowledge about the biosynthesis of PHA with a controlled composition from plant oils allowing in the future establishing the production of these polyesters as tailor-made polymers.

Polyhydroxyalkanoate biosynthesis in Bacillus cereus SPV under varied limiting conditions and an insight into the biosynthetic genes involved.

AIMS: A new strain of Bacillus, Bacillus cereus SPV, was found to be capable of using a wide range of carbon sources for the production of polyhydroxyalkanoates (PHAs) (Valappil et al. 2007b). Limiting nutrient in the culture conditions is crucial for PHA production. In this study, B. cereus SPV was grown in different culture conditions with limitation of potassium, nitrogen, sulphur and phosphorous to establish the impact of nutritional limitation on PHA production. METHODS AND RESULTS: The PHA yields obtained were found to be 13.4, 38, 13.15 and 33.33% dcw for potassium, nitrogen, sulphur and phosphorus limitations, respectively. Gas chromatography-mass spectrometry analysis of the isolated polymers showed the presence of P(3HB) under nitrogen, sulphur and phosphate-limiting conditions and P(3HB-3HV) copolymer under potassium limiting conditions. This ability of B. cereus SPV to accumulate different PHA monomers from structurally unrelated carbon sources led to an interest in the molecular analysis of PHA biosynthesis in this organism. To achieve this, PCR was used to identify the polyhydroxyalkanoate biosynthetic genes in B. cereus SPV. CONCLUSION: Sequence analysis of the PCR products from B. cereus SPV revealed the sequence of the putative biosynthetic genes, and possible regions involved in substrate binding. The nucleotide sequence reported in this paper is in the GenBank nucleotide sequence database under accession number DQ486135. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report comparing the capability of B. cereus SPV to produce PHAs under different culture conditions of potassium, nitrogen, sulfur and phosphate limitations. The results in this study suggest the unique ability of B. cereus SPV to supply both 3HB and 3HV monomers from a structurally unrelated carbon source, glucose.