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1.
Sebastian L. Riedel Christopher J. Brigham Charles F. Budde Johannes Bader ChoKyun Rha Ulf Stahl Anthony J. Sinskey 《Biotechnology and bioengineering》2013,110(2):461-470
Reduced downstream costs, together with high purity recovery of polyhydroxyalkanoate (PHA), will accelerate the commercialization of high quality PHA‐based products. In this work, a process was designed for effective recovery of the copolymer poly(hydroxybutyrate‐co‐hydroxyhexanoate) (P(HB‐co‐HHx)) containing high levels of HHx (>15 mol%) from Ralstonia eutropha biomass using non‐halogenated solvents. Several non‐halogenated solvents (methyl isobutyl ketone, methyl ethyl ketone, and butyl acetate and ethyl acetate) were found to effectively dissolve the polymer. Isoamyl alcohol was found to be not suitable for extraction of polymer. All PHA extractions were performed from both dry and wet cells at volumes ranging from 2 mL to 3 L using a PHA to solvent ratio of 2% (w/v). Ethyl acetate showed both high recovery levels and high product purities (up to 99%) when using dry cells as starting material. Recovery from wet cells, however, eliminates a biomass drying step during the downstream process, potentially saving time and cost. When wet cells were used, methyl isobutyl ketone (MIBK) was shown to be the most favorable solvent for PHA recovery. Purities of up to 99% and total recovery yields of up to 84% from wet cells were reached. During polymer recovery with either MIBK or butyl acetate, fractionation of the extracted PHA occurred, based on the HHx content of the polymer. PHA with higher HHx content (17–30 mol%) remained completely in solution, while polymer with a lower HHx content (11–16 mol%) formed a gel‐like phase. All PHA in solution could be precipitated by addition of threefold volumes of n‐hexane or n‐heptane to unfiltered PHA solutions. Effective recycling of the solvents in this system is predicted due to the large differences in the boiling points between solvent and precipitant. Our findings show that two non‐halogenated solvents are good candidates to replace halogenated solvents like chloroform for recovery of high quality PHA. Biotechnol. Bioeng. 2013; 110: 461–470. © 2012 Wiley Periodicals, Inc. 相似文献
2.
Polyhydroxyalkanoates (PHAs) are biopolyesters produced by microorganisms that are environmentally friendly. PHAs can be used to replace traditional plastic to reduce environmental pollution in various fields. PHA production costs are high because PHA must be produced from a carbon substrate. The purpose of this study was to find the strain that can used the BDF by-product as the sole carbon source to produce high amounts of medium-chain-length PHA. Three isolates were evaluated for potential PHA production by using biodiesel-derived crude glycerol as the sole carbon source. Among them, Pseudomonas mosselii TO7 yielded high PHA content. The PHA produced from P. mosselii TO7 were medium-chain-length-PHAs. The PHA content of 48% cell dry weight in 48 h with a maximum PHA productivity of 13.16 mg PHAs L?1 h?1. The narrow polydispersity index value of 1.3 reflected the homogeneity of the polymer chain, which was conducive to industrial applications. 相似文献
3.
Yung-Hun Yang Christopher J. Brigham Charles F. Budde Paolo Boccazzi Laura B. Willis Mohd Ali Hassan Zainal Abidin Mohd Yusof ChoKyun Rha Anthony J. Sinskey 《Applied microbiology and biotechnology》2010,87(6):2037-2045
We employed systematic mixture analysis to determine optimal levels of acetate, propionate, and butyrate for cell growth and
polyhydroxyalkanoate (PHA) production by Ralstonia eutropha H16. Butyrate was the preferred acid for robust cell growth and high PHA production. The 3-hydroxyvalerate content in the
resulting PHA depended on the proportion of propionate initially present in the growth medium. The proportion of acetate dramatically
affected the final pH of the growth medium. A model was constructed using our data that predicts the effects of these acids,
individually and in combination, on cell dry weight (CDW), PHA content (%CDW), PHA production, 3HV in the polymer, and final
culture pH. Cell growth and PHA production improved approximately 1.5-fold over initial conditions when the proportion of
butyrate was increased. Optimization of the phosphate buffer content in medium containing higher amounts of butyrate improved
cell growth and PHA production more than 4-fold. The validated organic acid mixture analysis model can be used to optimize
R. eutropha culture conditions, in order to meet targets for PHA production and/or polymer HV content. By modifying the growth medium
made from treated industrial waste, such as palm oil mill effluent, more PHA can be produced. 相似文献
4.
The CO-resistant strain B5786 of the hydrogen-oxidizing bacterium Alcaligenes eutrophuswas found to be able to synthesize polyhydroxyalkanoates (PHAs) under the conditions of growth limitation by nitrogen deficiency (the factor that promotes PHA synthesis) and growth inhibition by carbon monoxide. The gas mixtures that contained from 5 to 20 vol % CO did not inhibit the key enzymes of PHA synthesis–-ketothiolase, acetoacetyl-CoA reductase, hydroxybutyrate dehydrogenase, and PHA synthase. In the presence of CO, cells accumulated up to 70–75 wt % PHA (with respect to the dry biomass) without any noticeable increase in the consumption of the gas substrate. Chromatographic–mass spectrometric analysis showed that the PHA synthesized by A. eutrophusis a copolymer containing more than 99 mol % -hydroxybutyrate and trace amounts of -hydroxyvalerate. The PHA synthesized under the conditions described did not differ from that synthesized by A. eutrophuscells from electrolytic hydrogen. 相似文献
5.
The production of poly--hydroxyalkanoate (PHA) in Syntrophomonas wolfei grown in pure culture or in coculture with Methanospirillum hungatei was studied. PHA was produced by S. wolfei during the exponential phase of growth under both of these cultural conditions. S. wolfei in pure culture also produced PHA in stationary phase when the medium was supplemented with high concentrations of the substrate crotonate. In S. wolfei, PHA levels decreased after growth stopped and most of the substrate was depleted. Altering the C to N ratio of the medium did not affect the amount of PHA made per mg of protein. The incorporation of labeled butyrate but not acetate into PHA during the early stages of growth of S. wolfei and the fact that some of the PHA in a pure culture of S. wolfei grown with trans-2-pentenoate was a polymer of 5-carbon monomer units indicated that a pathway exists for the synthesis of PHA without degradation of the substrate to acetyl-CoA. During the later stages of growth, PHA was made by a pathway which was in equilibrium with the acetate pool. These data indicate that PHA served as a carbon/energy reserve material in S. wolfei, but the synthesis of this polymer was regulated in a manner different from the known pathways. 相似文献
6.
A. N. Boyandin S. V. Prudnikova M. L. Filipenko E. A. Khrapov A. D. Vasil’ev T. G. Volova 《Applied Biochemistry and Microbiology》2012,48(1):28-36
Biodegradation of microbial linear polymers of hydroxyalkanoic acids (polyhydroxyalkanoates, PHAs) by soil microbial communities
of different structures has been studied during two field seasons in different weather conditions. This process was shown
to be influenced by the polymer chemical composition, temperature, humidity, and the microbial soil component. The PHA degradation
was accompanied by a decrease in the polymer molecular weight and an increase in the degree of crystallinity, indicating the
preferential destruction of the amorphous phase compared to the crystalline one. The quantity of the true PHA destructors
developing at the surface of the polymer samples was lower than the quantity of accompanying bacteria. The dominant PHA degrading
microorganisms under the test conditions were identified as bacteria of the genera Variovorax, Stenotrophomonas, Acinetobacter, Pseudomonas, Bacillus, and Xanthomonas and as micromycetes from Penicillium, Paecilomyces, Acremonium, Verticillium, and Zygosporium. 相似文献
7.
Helmut Brandl Richard A. Gross Robert W. Lenz Ramona Lloyd R. Clinton Fuller 《Archives of microbiology》1991,155(4):337-340
In recent years industrial interest has been focussed on the evaluation of poly(3-hydroxyalkanoates) (PHA) as potentially biodegradable plastics for a wide range of technical applications. Studies have been carried out in order to optimize growth and culture conditions for the intracellular formation of PHA in the phototrophic, purple, non-sulfur bacterium Rhodobacter sphaeroides. Its potential to produce polyesters other than poly(3-hydroxybutyrate) (PHB) was investigated. On an industrial scale, the use of photosynthetic bacteria could harness sunlight as an energy source for the production of these materials. R. sphaeroides was grown anaerobically in the light on different carbon sources. Under nitrogenlimiting conditions a PHA content of up to 60 to 70% of the cellular dry weight was detected. In all of the cases studied, the storage polymer contained approximately 98 mol% of 3-hydroxybutyrate (HB) and 2 mol% 3-hydroxyvalerate (HV) monomer units. Decreasing light intensities did not stimulate PHA formation. Compared to Rhodospirillum rubrum (another member of the family of Rhodospirillaceae), R. sphaeroides showed a limited flexibility in its ability to form PHA with varying monomer unit compositions. 相似文献
8.
Bee-Yong Tay Bhadravathi Eswara Lokesh Chow-Yang Lee Kumar Sudesh 《World journal of microbiology & biotechnology》2010,26(6):1015-1024
The continuous quest for bacterial strains capable of accumulating polyhydroxyalkanoate (PHA) utilizing cheaper and renewable
carbon source prompted us to explore newer and diverse environments like the gut of termites. Among the bacterial strains
isolated from the gut of higher termite Macrotermes carbonarius, three strains were found to accumulate PHA, as observed by microscopic studies and PHA production experiments. Among them,
strain MC1 with rapid growth and higher PHA accumulation was selected for further studies. API kit-50 CHB and 16S rRNA gene
sequence analysis results indicated the strain to have 99% homology with Bacillus megaterium and Bacillus flexus.
Bacillus sp. MC1 was able to accumulate PHA during the growth phase utilizing different carbon sources like glucose, fructose, sodium
acetate, sodium valerate and 1,4-butanediol. Gas chromatography analysis of the polymer has shown it to be basically composed
of poly (3-hydroxybutyrate) (PHB). Growth associated PHB biosynthesis was best in the presence of sodium acetate with 39 wt%
after 16 h of cultivation. Though previous studies provided evidence confirming the presence of PHA producing bacteria in
termite gut, isolation and characterization of these strains in pure culture has not been documented yet. Presence of other
morphotypes in the termite gut with PHA like granular inclusions was evident from the transmission electron microscopy studies.
This is a novel report and shows the feasibility of using potent strains capable of utilizing lignocellulosic degradation
products as a renewable carbon source for the production of PHA in the future. 相似文献
9.
Wang Qian Zhang Hanxing Chen Quan Chen Xiulan Zhang Yuzhong Qi Qingsheng 《World journal of microbiology & biotechnology》2010,26(6):1149-1153
A deep-sea psychrotrophic bacterium Pseudoaltermonas sp. SM9913, isolated from abyssalbenthic sediments in Bohai Sea, was found to be able to synthesize polyhydroxyalkanoate (PHA)
polymer consisting of mainly 3-hydroxydecanoate and 3-hydroxydodecanoate. PHA accumulation in Pseudoaltermonas sp. SM9913 was confirmed by transmission electron microscope, Nile red dye and gas chromatography/mass spectral. The PHA content
was determined as high as 2–3% of cell dry weight when the strain was grown in a sea water-based liquid medium. The relationship
between PHA and exopolysaccharide accumulation in this strain was discussed. 相似文献
10.
Atlić A Koller M Scherzer D Kutschera C Grillo-Fernandes E Horvat P Chiellini E Braunegg G 《Applied microbiology and biotechnology》2011,91(2):295-304
Poly(hydroxyalkanoates) (PHAs) constitute biodegradable polyesters and are considered among the most promising candidates
to replace common petrochemical plastics in various applications. To date, all commercial processes for PHA production employ
microbial discontinuous fed-batch fermentations. These processes feature drawbacks such as varying product quality and the
inevitable periods of downtime for preparation and post-treatment of the bioreactor equipment. An unprecedented approach to
PHA production was chosen in the presented work using a multistage system consisting of five continuous stirred tank reactors
in series (5-SCR), which can be considered as a process engineering substitute of a continuous tubular plug flow reactor.
The first stage of the reactor cascade is the site of balanced bacterial growth; thereafter, the fermentation broth is continuously
fed from the first into the subsequent reactors, where PHA accumulation takes place under nitrogen-limiting conditions. Cupriavidus necator was used as production strain. The focus of the experimental work was devoted to the development of a PHA production process
characterized by high productivity and high intracellular polymer content. The results of the experimental work with the reactor
cascade demonstrated its potential in terms of volumetric and specific productivity (1.85 g L−1 h−1 and 0.100 g g−1 h−1, respectively), polymer content (77%, w/w) and polymer properties (M
w = 665 kg/mol, PDI = 2.6). Thus, implementing the technology for 5-SCR production of PHB results in an economically viable
process. The study compares the outcome of the work with literature data from continuous two-stage PHA production and industrial
PHA production in fed-batch mode. 相似文献
11.
The non-conventional yeast Arxula adeninivorans was equipped with the genes phbA, phbB and phbC of the polyhydroxyalkanoate (PHA) biosynthetic pathway of Ralstonia eutropha, which encode -ketothiolase, NADPH-linked acetoacetyl-CoA reductase and PHA synthase, respectively. Arxula strains transformed solely with the PHA synthase gene (phbC) were able to produce PHA. However, the maximum content of the polymer detected in these strains was just 0.003% poly-3-hydroxybutyrate (PHB) and 0.112% poly-3-hydroxyvalerate (PHV). The expression of all three genes (phbA, phbB, phbC) resulted in small increases in the PHA content of the transgenic Arxula cells. However, under controlled cultivation conditions with minimal medium and ethanol as the carbon source, the recombinant yeast was able to accumulate up to 2.2% PHV and 0.019% PHB. Possible reasons for these differences are discussed. 相似文献
12.
Buchnericin-LB adsorbs to gram-positive but not to gram-negative bacteria. The tested gram-positive bacteria were species of Lactobacillus, Pediococcus, Leuconostoc, Enterococcus, Lactococcus, Listeria, Bacillus, Staphylococcus; gram-negative bacteria belonged to the genera Salmonella, Escherichia, Yersinia and Pseudomonas. Buchnericin-LB adsorption depended on pH but not on time and temperature. Also some anions of salts and lipoteichoic acid reduced or inhibited its adsorption. Treatment of cells and cell walls of sensitive bacteria with detergents, organic solvents or enzymes did not affect subsequent binding of buchnericin-LB. Treatment with buchnericin-LB caused sensitive cells to lose high amounts of intracellular K+ ions and UV-absorbing materials and became more permeable to o-nitrophenol-β-D-galactopyranoside. Buchnericin-LB (640-2560 AU/ml) decreased the colony forming units (99%) and absorbance values of Listeria monocytogenes and Bacillus cereus. These results indicate that the mode of action of buchnericin-LB is bactericidal and its lethal effect is very rapid. 相似文献
13.
Aim: To boost short‐chain‐length‐long‐chain‐length polyhydroxyalkanoate (SCL‐LCL‐PHA) co‐polymer yield in Pseudomonas aeruginosa MTCC 7925 by manipulating culture conditions. Methods and Results: Stationary phase culture of P. aeruginosa MTCC 7925 was subjected to various doses of different carbons and, N and P deficiencies. Enhanced co‐polymer yield was recorded under ethanol‐ and glucose‐supplemented cultures. Interaction of ethanol with N‐deficiency boosted co‐polymer accumulation maximally. Conclusions: The sludge‐isolated P. aeruginosa MTCC 7925 demonstrated good capability to synthesize SCL‐LCL‐PHA co‐polymer from unrelated carbon sources. Supplementation of ethanol under N‐deficiency boosted the co‐polymer yield up to 69% of dry cell weight, which is significantly higher when compared with other SCL‐MCL‐PHA co‐polymer accumulating bacterial species. Significance and Importance of the Study: This is the first report on P. aeruginosa MTCC 7925, which is capable of accumulating LCL 3‐hydroxyhexadecanoate and 3‐hydroxyoctadecanoate units with SCL 3‐hydroxybutyrate and 3‐hydroxyvalerate as constituents of PHAs. This opens up new possibilities for various industrial applications owing to the superior properties of this new co‐polymer. 相似文献
14.
15.
Tobella LM Bunster M Pooley A Becerra J Godoy F Martínez MA 《Journal of industrial microbiology & biotechnology》2005,32(9):397-401
Poly-β-hydroxyalkanoates (PHA) polymer is synthesized by different bacterial species. There has been considerable interest
in the development and production of biodegradable polymers; however, the high cost of PHA production has restricted its applications.
Kraft cellulose industry effluents containing 2,4,6-trichlorophenol (10 or 20 μg ml−1) were used by the bacteria Sphingopyxis chilensis S37 and Wautersia sp. PZK to synthesize PHA. In this condition, S. chilensis S37 was able to grow and degrade 2,4,6-trichlorophenol (ca. 60%) and 80% of these cells accumulated PHA. Wautersia PZK completely degraded 2,4,6-TCP and more than 90% of the cells accumulated PHA in 72 h. The PHA detection was performed
by flow cytometry and polyester composition was characterized by gas chromatography-mass spectroscopy (GC-MS), indicating
that these polymers are made by 3-hydroxybutyric acid and 3-hydroxyhexadecanoic acid for S37 and PZK strains, respectively.
Results demonstrated that strains’ growth and PHA production and composition are not modified in cellulose effluents with
or without 2,4,6-TCP (10–20 μg ml−1). Therefore, our results indicate that S. chilensis S37 and Wautersia sp. PZK are able to degrade a toxic compound such as a 2,4,6-TCP and simultaneously produce a valuable biopolymer using low-value
substrates. 相似文献
16.
Christian Capello Stefanie Hellweg Konrad Hungerbühler 《Journal of Industrial Ecology》2008,12(1):111-127
A comparison of various waste‐solvent treatment technologies, such as distillation (rectification) and incineration in hazardous‐waste‐solvent incinerators and cement kilns, is presented for 45 solvents with respect to the environmental life‐cycle impact. The environmental impact was calculated with the ecosolvent tool that was previously described in Part I of this work. A comprehensive sensitivity analysis was performed, and uncertainties were quantified by stochastic modeling in which various scenarios were considered. The results show that no single treatment technology is generally environmentally superior to any other but that, depending on the solvent mixture and the process conditions, each option may be optimal in certain cases. Nevertheless, various rules of thumb could be derived, and a results table is presented for the 45 solvents showing under which process conditions and amount of solvent recovery distillation is environmentally superior to incineration. On the basis of these results and the ecosolvent tool, an easily usable framework was developed that helps decision makers in chemical industries reduce environmental burdens throughout the solvent life cycle. With clear recommendations on the environmentally optimized waste‐solvent treatment technology, the use of this framework contributes to more environmentally sustainable solvent management and thus represents a practical application of industrial ecology. 相似文献
17.
Virginia Martínez Edouard Jurkevitch José Luis García María Auxiliadora Prieto 《Environmental microbiology》2013,15(4):1204-1215
Bdellovibrio bacteriovorus HD100 is an obligate predator that invades and grows within the periplasm of Gram‐negative bacteria, including mcl‐polyhydroxyalkanoate (PHA) producers such as Pseudomonas putida. We investigated the impact of prey PHA content on the predator fitness and the potential advantages for preying on a PHA producer. Using a new procedure to control P. putida KT2442 cell size we demonstrated that the number of Bdellovibrio progeny depends on the prey biomass and not on the viable prey cell number or PHA content. The presence of mcl‐PHA hydrolysed products in the culture supernatant after predation on P. putida KT42Z, a PHA producing strain lacking PhaZ depolymerase, confirmed the ability of Bdellovibrio to degrade the prey's PHA. Predator motility was higher when growing on PHA accumulating prey. External addition of PHA polymer (latex suspension) to Bdellovibrio preying on the PHA minus mutant P. putida KT42C1 restored predator movement, suggesting that PHA is a key prey component to sustain predator swimming speed. High velocities observed in Bdellovibrio preying on the PHA producing strain were correlated to high intracellular ATP levels of the predator. These effects brought Bdellovibrio fitness benefits as predation on PHA producers was more efficient than predation on non‐producing bacteria. 相似文献
18.
A novel lipase-catalyzed synthesis of aliphatic polyesters has been achieved successfully by using Candida sp.99-125 with diethyl sebacate and 1,4-butanediol as starting substrates in absence of organic solvents. The lipase from Candida sp.99-125 was employed for the first time to catalyze synthesis of poly(butylene sebacate) and showed high catalytic activity for bulkpolymerization under mild reaction conditions. The eco-friendly processes, without any environmental pollution, avoided both phase separation of the reactants and the use of toxic solvents effectively. The polycondensation reactions were performed in two stages: first stage reaction under atmospheric pressure to convert the monomers to oligomers followed by second stage reaction under vacuum process, with a highest molecular weight of 15,800 obtained in molar ratio of 1:1 at 70 °C. The corresponding polyesters determined by thermogravimetric analysis (TGA) were much thermally stable up to 300 °C. Differential scanning calorimetry (DSC) and wide-angle X-ray diffraction (WAXS) analysis demonstrated the polymers had a strong ability to crystallize with the degree of crystallinity up to 66%. 相似文献
19.
Clemente T Shah D Tran M Stark D Padgette S Dennis D Brückener K Steinbüchel A Mitsky T 《Applied microbiology and biotechnology》2000,53(4):420-429
A 3.0-kb genomic fragment has been isolated from Rhodospirillum rubrum (ATCC 25903) that contains an open reading frame (ORF) with strong homology to other known polyhydroxyalkanoate (PHA) synthase
genes. This ORF has lower homology to the R. rubrum strain Ha PHA synthase than would be expected within the same species. We have conducted a series of heterologous expression
studies evaluating the in vivo substrate specificity of PHA synthase genes from Rhodobacter sphaeroides, Ralstonia eutropha (formerly Alcaligenes eutrophus), Thiocystis violacea, and Nocardia corrallina, within the PHA-synthase-negative hosts, Ralstonia eutropha DSM541 and Pseudomonas putida GpP104. The N. corrallina PHA synthase incorporated the highest percentage of C5 monomers in the polymer when fermented in medium supplemented with
0.1% heptanoate as the sole carbon source. When the T. violacea and R. sphaeroides were expressed in the PHA-negative host DSM541, a greater percentage of C5 monomer was observed in the polymer as compared
to the expression of the PHA synthase of R. eutropha, when the transconjugants were fermented in medium supplemented with 0.4% propionate. Evaluation for preference of medium-chain-length
monomers demonstrated the flexibility of the N. corrallina, T. violacea, and R. eutropha synthase genes to polymerize a copolyester composed of short- and medium-chain-length monomers when the respective transconjugants
were fermented in medium supplemented with 0.5% octanoate. These studies demonstrate that the PHA synthase from N. corrallina, T. violacea, and R. eutropha are able to polymerize a copolyester composed of short- and medium-chain-length monomers, while the PHA synthase from R. sphaeroides lacks this ability and only produces a short-chain-length polymer. These observations suggest that the composition of the
PHA from the PHA-producing organisms does not necessarily reflect the inherent specificity of the PHA synthase.
Received: 16 March 1999 / Received revision: 24 August 1999 / Accepted: 24 September 1999 相似文献
20.
Conjugated Polymers Based on Difluorobenzoxadiazole toward Practical Application of Polymer Solar Cells
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Junyi Wang Shiliang Wang Chunhui Duan Fallon J. M. Colberts Jiangquan Mai Xi Liu Xiao'e Jia Xinhui Lu René A. J. Janssen Fei Huang Yong Cao 《Liver Transplantation》2017,7(22)
To advance polymer solar cells (PSCs) toward real‐world applications, it is crucial to develop materials that are compatible with a low‐cost large‐scale manufacturing technology. In this context, a practically useful polymer should fulfill several critical requirements: the capability to provide high power conversion efficiencies (PCEs) via low‐cost fabrication using environmentally friendly solvents under mild thermal conditions, resulting in an active layer that is thick enough to minimize defects in large‐area films. Here, the development of new photovoltaic polymers is reported through rational molecular design to meet these requirements. Benzodithiophene (BDT)‐difluorobenzoxadiazole (ffBX)‐2‐decyltetradecyl (DT), a wide‐bandgap polymer based on ffBX and BDT emerges as the first example that fulfills the qualifications. When blended with a low‐cost acceptor (C60‐fullerene derivative), BDT‐ffBX‐DT produces a PCE of 9.4% at active layer thickness over 250 nm. BDT‐ffBX‐DT devices can be fabricated from nonhalogenated solvents at low processing temperature. The success of BDT‐ffBX‐DT originates from its appropriate electronic structure and charge transport characteristics, in combination with a favorable face‐on orientation of the polymer backbone in blends, and the ability to form proper phase separation morphology with a fibrillar bicontinuous interpenetrating network in bulk‐heterojunction films. With these characteristics, BDT‐ffBX‐DT represents a meaningful step toward future everyday applications of polymer solar cells. 相似文献