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Conversion of industrial food wastes by Alcaligenes latus into polyhydroxyalkanoates.

Broader usage of biodegradable plastics in packaging and disposable products as a solution to environmental problems would heavily depend on further reduction of costs and the discovery of novel biodegradable plastics with improved properties. As the first step in our pursuit of eventual usage of industrial food wastewater as nutrients for microorganisms to synthesise environmental-friendly bioplastics, we investigated the usage of soya wastes from a soya milk dairy, and malt wastes from a beer brewery plant as the carbon sources for the production of polyhydroxyalkanoates (PHA) by selected strain of microorganism. Bench experiments showed that Alcaligenes latus DSM 1124 used the nutrients from malt and soya wastes to biosynthesise PHAs. The final dried cell mass and specific polymer production of A. latus DSM 1124 were 32g/L and 70% polymer/cells (g/g), 18.42 g/L and 32.57% polymer/cell (g/g), and 28 g/L and 36% polymer/cells (g/g), from malt waste, soya waste, and from sucrose, respectively. These results suggest that many types of food wastes might be used as the carbon source for the production of PHA.

Journal Article↗

Biodegradation of plasticizers by Rhodococcus rhodochrous.

Rhodococcus rhodochrous was grown in the presence of one of three plasticizers: bis 2-ethylhexyl adipate (BEHA), dioctyl phthalate (DOP) or dioctyl terephthalate (DOTP). None of the plasticizers were degraded unless another carbon source, such as hexadecane, was also present. When R. rhodochrous was grown with hexadecane as a co-substrate, BEHA was completely degraded and the DOP was degraded slightly. About half of the DOTP was degraded, if hexadecane were present. In all of these growth studies, the toxicity of the media, which was assessed using the Microtox assay, increased as the organism degraded the plasticizer. In each case, there was an accumulation of one or two intermediates in the growth medium as the toxicity increased. One of these was identified as 2-ethylhexanoic acid and it was observed for all three plasticizers. Its concentration increased until degradation of the plasticizers had stopped and it was always present at the end of the fermentation. The other intermediate was identified as 2-ethylhexanol and this was only observed for growth in the presence of BEHA. The alcohol was observed early in the growth studies with BEHA and had disappeared by the end of the experiment. Both the 2-ethylhexanol and 2-ethylhexanoic acid were shown to be toxic and their presence explained the increase of toxicity as the fermentations proceeded. The appearance of these intermediates was consistent with similar degradation mechanisms for all three plasticizers involving hydrolysis of the ester bonds followed by oxidation of the released alcohol.

Alkanes↗

PHA (polyhydroxyalkanoate) production potential of activated sludge treating wastewater.

The main purposes of wastewater treatment systems are to remove organic pollutants, but it would be very attractive if there were a way to recover the organic pollutants as valuable organic materials. One of the possible ways to recover organic pollutants in wastewater is to convert them into polyhydroxyalkanoates (PHAs), which are biodegradable plastics. In this study, 18 activated sludge samples in 4 wastewater treatment plants (WWTPs) in Tokyo, Japan, were evaluated for their potential to produce PHAs by aerobic batch experiments with excess supply of acetate as the sole carbon source. The activated sludge samples tested had the capability to accumulate PHA up to 18.8% of dry cell weight on average, with the range of 6.0% to 29.5%. The results showed that the maximum PHA content was dependent on the influent more than on the operational conditions of the activated sludge, and that conventional activated sludge produced PHA as much as anaerobic-aerobic activated sludge did. The PHA content achieved in this study is still low, and further improvement is needed to put into practice the recovery process of organic pollutants as biodegradable plastics by activated sludge.

Alkanes↗

Biodegradation of plasticizers by Rhodotorula rubra.

The degradation of plasticizers by the yeast Rhodotorula rubra J-96-1 (American Type Culture Collection 9449) in the presence of glucose was studied. The plasticizers included the commonly used bis-2-ethylhexyl adipate (B(EH)A), dioctyl phthalate (DOP), and dioctyl terephthalate (DOTP), and the less commonly used dipropylene glycol dibenzoate (D(PG)DB) and diethylene glycol dibenzoate (D(EG)DB). The proposal had been made that the latter two plasticizers be used as alternatives to the first three, which have been associated with negative environmental impacts. The degradation of D(PG)DB or D(EG)DB led to a significant increase in solution toxicity, which was associated with the production of metabolites resulting from the incomplete breakdown of the plasticizers. The toxic metabolites in the D(PG)DB system were identified as isomers of dipropylene glycol monobenzoate. A pathway for the formation of this metabolite was proposed. The metabolite observed when D(EG)DB was being degraded was tentatively identified as diethylene glycol monobenzoate by analogy to the D(PG)DB system. In contrast, no metabolites were observable and toxicity did not increase in the media during the degradation of B(EH)A, DOP, or DOTP by R. rubra. Collectively, these results do not support the use of D(PG)DB and D(EG)DB as environmentally safe alternatives to B(EH)A, DOP, or DOTP.

Benzoates↗

Environmentally degradable, high-performance thermoplastics from phenolic phytomonomers.

Aliphatic polyesters, such as poly(lactic acid), which degrade by hydrolysis, from naturally occurring molecules form the main components of biodegradable plastics. However, these polyesters have become substitutes for only a small percentage of the currently used plastic materials because of their poor thermal and mechanical properties. Polymers that degrade into natural molecules and have a performance closer to that of engineering plastics would be highly desirable. Although the use of a high-strength filler such as a bacterial cellulose or modified lignin greatly increases the plastic properties, it is the matrix polymer that determines the intrinsic properties of the composite. The introduction of an aromatic component into the thermoplastic polymer backbone is an efficient method to intrinsically improve the material performance. Here, we report the preparation of environmentally degradable, liquid crystalline, wholly aromatic polyesters. The polyesters were derived from polymerizable plant-derived chemicals--in other words, 'phytomonomers' that are widely present as lignin biosynthetic precursors. The mechanical performance of these materials surpasses that of current biodegradable plastics, with a mechanical strength, sigma, of 63 MPa, a Young's modulus, E, of 16 GPa, and a maximum softening temperature of 169 degrees C. On light irradiation, their mechanical properties improved further and the rate of hydrolysis accelerated.

Journal Article↗

Production of organic acids from kitchen wastes.

This study involves the production of short-chain organic acids from kitchen wastes as intermediates for the production of biodegradable plastics. Flasks, without mixing were used for the anaerobic conversion of the organic fraction of kitchen wastes into short-chain organic acids. The influence of pH, temperature and addition of sludge cake on the rate of organic acids production and yield were evaluated. Fermentations were carried out in an incubator at different temperatures controlled at 30 degrees C. 40 degrees C, 50 degrees C, 60 degrees C and uncontrolled at room temperature. The pH was also varied at pH 5, 6, 7, and uncontrolled pH. 1.0 M phosphate buffer was used for pH control, and 1.0 M HCl and 1.0 M NaOH were added when necessary. Sludge cake addition enhanced the rate of maximum acids production from 4 days to 1 day. The organic acids produced were maximum at pH 7 and 50 degrees C i.e., 39.84 g/l on the fourth day of fermentation with a yield of 0.87 g/g soluble COD consumed, and 0.84 g/g TVS. The main organic acid produced was lactic acid (65-85%), with small amounts of acetic (10-30%), propionic (5-10%), and butyric (5-20%) acids. The results of this study showed that kitchen wastes could be fermented to high concentration of organic acids, which could be used as substrates for the production of biodegradable plastics.

Anaerobiosis↗

Production of L-lactic acid from fresh cassava roots slurried with tofu liquid waste by Streptococcus bovis.

To reduce the production cost of biodegradable plastics, the fermentation performance of L-lactic acid for a new fermentation medium, fresh cassava roots (FCRs) as a substrate slurried with tofu liquid waste (TLW) as basal medium, was investigated by batch fermentation of Streptococcus bovis. The fermentation properties of the three substrates, namely, FCR, tapioca (cassava starch) and glucose, which were independently mixed with TLW, were compared with those independently mixed with the standard basal medium, trypto-soya broth (TSB). Experiments were conducted at various sugar concentrations of the substrates with CaCO(3) as a neutralizer. The maximum L-lactic acid concentrations (C(La)) obtained using the three substrates in TLW were about 75% of those obtained using TSB caused by less nutrients in the TLW. The L-lactic acid productivities (P(La)) and the specific growth rates of S. bovis (mu) in TLW were about 1/4 to 1/3 and 1/5 to 1/4 of those in TSB, respectively. The maximum C(La), P(La) and mu were obtained at 10% w/w sugar concentration. Total yields (eta) were nearly constant up to 10% w/w sugar concentration for TSB and TLW, that is, 80% to 85% and 50% to 60%, respectively. But their total yields decreased in more than 10% w/w sugar concentration in both basal media, because of substrate inhibition. The fermentation properties (C(La), P(La), mu, and eta) were found to be in the order of: FCR > tapioca > glucose for all concentrations of the three substrates. The fermentation properties for FCR and tapioca were higher than those for glucose, in TLW or TSB, because S. bovis in a medium containing starch (FCR and tapioca) has more amylase activity than in a medium containing glucose. The nutrients in FCR with poor nutrient basal medium (TLW) more strongly affected the fermentation properties than those in FCR with rich nutrient basal medium (TSB). The proposed fermentation medium of FCR slurried with TLW is worth studying in order to reduce production cost of biodegradable plastics.

Biotransformation↗

Crystallinity in starch plastics: consequences for material properties.

The processing of starches with biodegradable additives has made biodegradable plastics suitable for a number of applications. Starch plastics are partially crystalline as a result of residual crystallinity and the recrystallization of amylose and amylopectin. Such crystallinity is a key determinant of the product's properties. This article describes the influence of processing and storage conditions on starch crystallinity and offers possible explanations for the various properties of starch plastics, in particular for the problems associated with ageing, in terms of the different crystalline structures.

Biodegradation, Environmental↗

Nature or petrochemistry?-biologically degradable materials.

Naturally occurring polymers have been utilized for a long time as materials, however, their application as plastics has been restricted because of their limited thermoplastic processability. Recently, the microbial synthesis of polyesters directly from carbohydrate sources has attracted considerable attention. The industrial-scale production of poly(lactic acid) from lactic acid generated by fermentation now provides a renewable resources-based polyester as a commodity plastic for the first time. The biodegradability of a given material is independent of its origin, and biodegradable plastics can equally well be prepared from fossil fuel feedstocks. A consideration of the overall carbon dioxide emissions and consumption of non-renewable resources over the entire life-cycle of a product is not necessarily favorable for plastics based on renewable resources with current technology-in addition to the feedstocks for the synthesis of the polymer materials, the feedstock for generation of the overall energy required for production and processing is decisive.

Bacteria↗

Priming effect as determined by adding 14C-glucose to modified controlled composting test.

The development of new biodegradable packaging materials, especially biodegradable plastics, has created a need for biodegradability testing. The European standard for controlled composting test was used in this study for assessing if the addition of a test material results in excess CO2 production in compost. This effect, designated as the priming effect, would give an erroneous result for biodegradation, which is based on CO2 formation from the test material. Glucose was selected as a test substrate because it is the degradation product of starch and cellulose, which are major compounds of many packaging materials. Both 14C-glucose and non-labelled glucose was applied to nine compost samples of variable stability and age from two weeks to 1.5 years. CO2 and 14CO2 evolution were measured during the incubation. Biodegradation of glucose in unstable composts (age <6 months) was negative and 14CO2 evolution was poor, although the respective composts without glucose produced relatively high amounts of CO2. It was concluded that a negative priming effect was observed in unstable composts, in which glucose remained mostly non-degraded and apparently inhibited the mineralization of native organic matter in the compost. In stable composts (age > or = 6 months), biodegradation of glucose was high and approximately equal to 14C-glucose mineralization, i.e., the composts showed no priming effect. Young composts were unsuitable for controlled composting test due to lack of stability. It is important to ensure that the compost inoculum used for the test is sufficiently stable.

Bacteria↗

Construction of recombinant Escherichia coli strains for production of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate).

Plastic wastes constitute a worldwide environmental problem, and the demand for biodegradable plastics has become high. One of the most important characteristics of microbial polyesters is that they are thermoplastic with environmentally degradable properties. In this study, pUC19/PHA was cloned and transformed into three different Escherichia coli strains. Among the three strains that were successfully expressed in the production of polyhydroxyalkanoates (PHA), E. coli HMS174 had the highest yield in the production of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) (P[HB-HV]). The cell dry weight and PHA content of recombinant HMS174 reached as high as 10.27 g/L and 43% (w/w), respectively, in fed-batch fermentor culture. The copolymer of PHA, P(HB-HV), was found in the cells, and the biopolymers accumulated were identified and analyzed by gas chromatography, proton nuclear magnetic resonance spectroscopy, and differential scanning calorimetry. We demonstrated clearly that the E. coli host for PHA production has to be carefully selected to obtain a high yield. The results obtained indicated that a superior E. coli with high PHA production can be constructed with a desirable ratio of P(HB-HV), which has potential applications in industry and medicine.

Biodegradation, Environmental↗

Production of polyhydroxybutyrate by Bacillus species isolated from municipal activated sludge.

Plastic wastes are considered to be severe environmental contaminants causing waste disposal problems. Widespread use of biodegradable plastics is one of the solutions, but it is limited by high production cost. Biologic wastewater treatment generates large quantities of biomass as activated sludge. Only a few reports focus on the potential of utilizing resident Bacillus species from activated sludge in polyhydroxbutyrate (PHB) production as well as the production of PHB from food wastes. They have attractive properties such as short generation time, absence of endotoxins, and secretion of both amylases and proteinases that can well utilize food wastes for nutrients, which can further reduce the cost of production of polyhydroxyalkanoates (PHAs). Two PHA-producing strains, HF-1 and HF-2, were isolated from activated sludge. HF-1 outperformed HF-2 in terms of growth and PHB production in hydrolyzed soy and malt wastes. The isolated bacteria was characterized by DNA sequence alignment. Cell extracts of HF-1 were also compared to Bacillus megaterium cell extracts on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The biopolymers accumulated were analyzed by gas chromatography, nuclear magnetic resonance, and Fourier transform infrared methods.

Bacillus↗

Materials derived from biomass/biodegradable materials.

Interest in biodegradable plastics made from renewable resources has increased significantly in recent years. PHBV (polyhydroxybutyrate-polyhydroxyvalerate) copolymers are good examples of this type of materials. This paper provides an overview of the manufacturing process, properties, biodegradability, and application/commercial issues associated with PHBV copolymers. They are naturally produced by bacteria from agricultural raw materials, and they can be processed to make a variety of useful products, where their biodegradability and naturalness are quite beneficial. PHBV copolymers are still in the first stage of commercialization. But they are presented in this paper as an example of how new technology can help meet society's needs for plastics and a clean environment.

Alcaligenes↗

Lipase-catalyzed transformation of poly(butylene adipate) and poly(butylene succinate) into repolymerizable cyclic oligomers.

The enzymatic degradation and repolymerization were carried out with the objectives of developing the chemical recycling of aliphatic polyester-type plastics, such as the poly(butylene adipate) (PBA), poly(butylene succinate), and poly(butylene adipate-co-succinate) copolymers which are typical biodegradable plastics. They were degraded by lipase in an organic solvent solution containing a small amount of water to produce cyclic oligomers mainly consisting of the cyclic diester. The produced cyclic oligomer was readily repolymerized by lipase to produce a polyester having an equal or higher molecular weight compared to the parent polymer. As an example, PBA having an Mw of 22,000 was almost quantitatively transformed by lipase CA (Novozym 435) in water-containing toluene at 50 degrees C into the corresponding cyclic oligomers mainly consisting of dimers. Thus, the obtained oligomers were readily polymerized by lipase CA to produce the PBA with an Mw of 52,000.

Adipates↗

Ability of the phototrophic bacterium Rhodospirillum rubrum to produce various poly (beta-hydroxyalkanoates): potential sources for biodegradable polyesters.

Studies have been carried out in order to optimize growth and culture conditions for the intracellular formation of poly(beta-hydroxyalkanoates) (PHA) in the phototrophic, purple, non-sulphur bacterium Rhodospirilum rubrum. Its potential to produce novel copolymers was investigated. Recently, it has become of industrial interest to evaluate these polyesters as potentially biodegradable plastics for a wide range of possible applications. On an industrial scale, the use of photosynthetic bacteria could harness sunlight as an energy source for the production of these materials. R. rubrum was grown anaerobically in the light on different linear and branched beta-hydroxycarboxylic acids and various n-alkanoic acids. Under nitrogen-limiting conditions a PHA content of up to 45% of cellular dry weight was detected. When R. rubrum was grown on different concentrations of various n-alkanoic acids, intracellular PHA production was detected on all acids used. In most of the cases, the storage polymer contained beta-hydroxybutyrate (HB) and beta-hydroxyvalerate (HV) monomer units. Grown on n-alkanoic acids with a chain length of four carbon atoms and more, R. rubrum produced a copolymer containing the beta-hydroxyhexanoate (HC) repeating unit in addition to the HB and HV monomer. Using beta-hydroxyheptanoic acid as the carbon source, a polyester which contained HB, HV, HC, and beta-hydroxyheptanoate was formed. These copolyesters represent a novel class of biodegradable thermoplastics. The results demonstrate the metabolic flexibility of R. rubrum to form many different types of polyesters which might substitute plastics synthesized from petrochemicals.

Ammonia↗

A two step chemo-biotechnological conversion of polystyrene to a biodegradable thermoplastic.

A novel approach to the recycling of polystyrene is reported here; polystyrene is converted to a biodegradable plastic, namely polyhydroxyalkanoate (PHA). This unique combinatorial approach involves the pyrolysis of polystyrene to styrene oil, followed by the bacterial conversion of the styrene oil to PHA by Pseudomonas putida CA-3 (NCIMB 41162). The pyrolysis (520 degrees C) of polystyrene in a fluidized bed reactor (Quartz sand (0.3-0.5 mm)) resulted in the generation of an oil composed of styrene (82.8% w/w) and low levels of other aromatic compounds. This styrene oil, when supplied as the sole source of carbon and energy allowed for the growth of P. putida CA-3 and PHA accumulation in shake flask experiments. Styrene oil (1 g) was converted to 62.5 mg of PHA and 250 mg of bacterial biomass in shake flasks. A 1.6-fold improvement in the yield of PHA from styrene oil was achieved by growing P. putida CA-3 in a 7.5 liter stirred tank reactor. The medium chain length PHA accumulated was comprised of monomers 6, 8, and 10 carbons in length in a molar ratio of 0.046:0.436:1.126, respectively. A single pyrolysis run and four fermentation runs resulted in the conversion of 64 g of polystyrene to 6.4 g of PHA.

Biodegradation, Environmental↗

Optimal production of polyhydroxyalkanoates in activated sludge biomass.

Polyhydroxyalkanoates (PHAs) have been recognized as good candidates for biodegradable plastics, but their high price compared with conventional plastics has limited their use. In this study, activated sludge microorganisms from a conventional wastewater treatment process were induced, by controlling the carbon:nitrogen (C:N) ratio in the reactor liquor, to accumulate PHAs. In addition, an intermittent nitrogen feeding program was established to optimize the volumetric PHA productivity in a wastewater treatment process. The optimal overall polymer production yield of 0.111 g of polymer/g of carbonaceous substrate consumed was achieved under a C:N ratio of 96:1 by feeding nitrogen in the reactor liquor once every four cycles. At the same time, the amount of excess sludge generated from the wastewater treatment process was reduced by 22.9%.

Bacteria↗