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Polyhydroxyalkanoate (PHA) production from waste.

PHA (polyhydroxyalkanoate) production was attempted with SBRs from food waste. Seed microbes were collected from a sewage treatment plant with a biological nutrient removal process, and acclimated with synthetic substrate prior to the application of the fermented food waste. Laboratory SBRs were used to produce PHA with limited oxygen and nutrients. The maximum content of 51% PHA was obtained with an anaerobic/aerobic cycle with P limitation, and the yield was estimated to be about 0.05 gPHA(produced)/gCOD(applied) or 25 kg PHA/dry ton of food waste, assuming more than 40% of the PHA contents were recoverable. PHB/PHA ratios were 0.74 to 0.77 due to the higher acetate concentrations. Economical analysis seemed to suggest the PHA produced from the food waste could be an alternative material to produce the biodegradable plastic to be used for the collection bags for solid waste.

Acetates↗

A rapid method for the determination of metal toxicity to the biodegradation of water insoluble polymers.

A rapid and inexpensive test has been developed to investigate the influence of elevated heavy metal concentrations on the degradation of biodegradable plastics. Using an overlay agar technique it could be demonstrated that copper concentrations above 8 to 14 mg/L inhibit the degradation of powdered poly(3-hydroxybutyrate), (PHB), by Acidovorax delafieldii. This is the first study demonstrating the inhibiting influence of metals on the biodegradation of PHB.

Journal Article↗

Evaluation of traps and toxicants in an attract-and-kill system for Rhagoletis mendax (Diptera: Tephritidae).

A series of laboratory and field assays were performed to evaluate new methods for an attract-and-kill approach to control blueberry maggot, Rhagoletis mendax Curran. In laboratory assays, fly mortality was similar among insecticide-treated matted paper, plastic, and biodegradable sphere traps. Plastic and biodegradable traps baited with ammonium acetate captured significantly more flies than baited matted paper traps in field assays, whereas fly captures were similar in baited matted paper traps and unbaited plastic sphere traps. Treatments containing imidacloprid resulted in the highest amount of fly knockdown, and spheres coated with this insecticide still had significant knockdown after 6 wk of field exposure. Fly mortality was comparable on sphere traps coated with fipronil and imidacloprid, with both resulting in significantly more fly mortality than the control when flies were exposed to traps that had been weathered for 4 wk in the field. Spheres coated with acetamiprid resulted in fly mortality, but further evaluation is necessary to determine the potential of this compound. Deltamethrin was ineffective in causing fly knockdown at the rate tested. Our results indicate that fipronil and imidacloprid have potential as insecticidal coatings on either plastic or biodegradable spheres in an attract-and-kill system for control of R. mendax.

Acetates↗

Development of biobased products.

Research conducted over the past seven years by the biotechnology byproducts consortium (BBC) addresses its mission to investigate the opportunities to add value to agricultural products, byproducts and coproducts and to manage the wastewater arising from agribusinesses in an environmentally favorable way. Since a wide variety of research approaches have been taken, the results are collected in five topic groups: (1) bioremediation that includes anaerobic fermentations of wastes to produce methane and hydrogen, the genetics of methanogenesis and in situ remediation of contaminated aquifer systems, landfill leachates and industrial effluents; (2) land application of fermentation byproducts and their use in animal feeds; (3) biocatalytic studies of transformations of components of corn and soybean oils, peroxidases present in plant products, such as soybean hulls; (4) biochemical reactions for the production of de-icers from industrial water streams, biodiesel production from fats and greases, biodegradable plastics from polymerizable sugar derivatives, single cell foods derived from fungal growth on waste streams, and bacterial polysaccharides from Erwinia species; (5) separation and recovery of components by membrane technologies.

Animal Feed↗

Production of polyhydroxyalkanoates (PHA) by activated sludge treating municipal wastewater: effect of pH, sludge retention time (SRT), and acetate concentration in influent.

In this paper, the production of biodegradable plastics polyhydroxyalkanoates (PHA) by activated sludge treating municipal wastewater was investigated. The effect of three operational factors, i.e. the acetate concentration in influent, pH, and sludge retention time (SRT) were studied. Sludge acclimatized with municipal wastewater supplemented with acetate could accumulate PHA up to 30% of sludge dry weight, while sludge acclimatized with only municipal wastewater achieved 20% of sludge dry weight. It was found that activated sludge with an SRT of 3 days possessed better PHA production capability than sludge with an SRT of 10 days. Sludge acclimatized under pH 7 and 8 conditions in sequencing batch reactors (SBRs) exhibited similar PHA production capability. However, in PHA production batch experiments, pH value influenced significantly the PHA accumulation behavior of activated sludge. When pH of batch experiments was controlled at 6 or 7, a very low PHA production was observed. The production of PHA was stimulated when pH was kept at 8 or 9.

Acetates↗

Transformation of biodegradable polyesters into cyclic oligomers under continuous flow using an enzyme-packed column.

The enzymatic degradation of typical chemically synthesized biodegradable plastics, such as poly(butylene adipate) and poly(epsilon-caprolactone), to a repolymerizable cyclic oligomer under continuous flow using an enzyme column was studied, directed towards green chemical recycling. It was confirmed that both polymers were quantitatively transformed into the corresponding cyclic oligomers by passage through the column packed with immobilized lipase from Candida antarctica (Novozym 435) at 40 degrees C using a toluene solution. Compared to the batch treatment in a flask, a short reaction time and a low reaction temperature occurred for complete transformations of the polyesters by continuous flow through the enzyme column. The degradation activity of the enzyme column was maintained for at least six months at 40 degrees C. Conceptual scheme of the continuous degradation equipment using the enzyme column.

Biocompatible Materials↗

Production of specific copolymers of polyhydroxyalkanoates from industrial waste.

Polyhydroxyalkanoates, biodegradable plastics with the desired physical and chemical properties of conventional synthetic plastics, are extensively investigated. In this study, specific bacterial strains produced specific copolymers from food waste. Copolymers of HB and HV (poly[3-hydroxybutyrate-co-3-hydroxyvalerate]) were obtained using various ratios of butyric acid (C4) and valeric acid (C5) as carbon sources. The C4 to C5 ratio affected the melting points of the copolymers. Melting and glass transition temperatures and many other thermal properties are important parameters relative to in-service polymer applications. Higher ratios of butyrate to valerate gave higher melting points. When a mixed culture of activated sludge was employed to produce copolymers using food wastes as nutrients, the obtained copolymers showed various monomer compositions. Copolymers with a higher portion of HV were obtained using soy waste; copolymers with less HV were obtained using malt wastes. Pure strains, (i.e., Alcaligenes latus DSM 1122, and DSM 1124, Staphylococcus spp., Klebsiella spp.) produced specific copolymers from food waste. Only Klebsiella spp. produced different copolymers; the ratios of HB:HV were 93:7 and 79:21 from malt waste and soy waste, respectively. The other strains produced polymers of 100% HB. Selecting industrial food wastes as carbon sources can further reduce the cost of producing copolymers.

Alcaligenes↗

Polyhydroxyalkanoates (PHA) production using wastewater as carbon source and activated sludge as microorganisms.

Activated sludge from different full-scale wastewater treatment plants (municipal, pulp and paper industry, starch manufacturing and cheese manufacturing wastewaters) was used as a source of microorganisms to produce biodegradable plastics in shake flask experiments. Acetate, glucose and different wastewaters were used as carbon sources. Pulp and paper wastewater sludge was found to accumulate maximum concentration (43% of dry weight of suspended solids) of polyhydroxy alkanoates (PHA) with acetate as carbon source. Among the different wastewaters tested as a source of carbon, pulp and paper industry and starch industry wastewaters were found to be the best source of carbon while employing pulp and paper activated sludge for maximum accumulation of PHA. High concentration of volatile fatty acids in these wastewaters was the probable reason.

Acetates↗

Production of PHB by a Bacillus megaterium strain using sugarcane molasses and corn steep liquor as sole carbon and nitrogen sources.

Poly(hydroxybutyric acid) (PHB) and other biodegradable polyesters are promising candidates for the development of environment-friendly, totally biodegradable plastics. The use of cane molasses and corn steep liquor, two of the cheapest substrates available in Egypt, may help to reduce the cost of producing such biopolyesters. In this work, the effect of different carbon sources was studied. Maximum production of PHB was obtained with cane molasses and glucose as sole carbon sources (40.8, 39.9 per mg cell dry matter, respectively). The best growth was obtained with 3% molasses, while maximum yield of PHB (46.2% per mg cell dry matter) was obtained with 2% molasses. Corn steep liquor was the best nitrogen source for PHB synthesis (32.7 mg per cell dry matter), on the other hand, best growth was observed when ammonium chloride, ammonium sulphate, ammonium oxalate or ammonium phosphate were used as nitrogen sources.

Bacillus megaterium↗

Batch production of polyhydroxyalkanoate by low-polyphosphate-content activated sludge at varying pH.

Polyhydroxyalkanoate (PHA), a biodegradable plastic, can be produced from excess activated sludge by utilizing intracellular glycogen and polyphosphate as energy sources under growth-limiting conditions. Activated sludge of 2%, 6%, and 8% polyphosphate with similar glycogen content of 33% was investigated for batch PHA production by varying the pH values from 6 to 8. Acetate applied at 1000 mg COD/L was almost exhausted within 80 min of anaerobic stage. The remaining glycogen in the sludge was higher at a lower pH because of less energy used for acetate uptake. Highest PHA content of 51% was obtained from sludge with an 8% polyphosphate content at pH 8. PHA production occurred rapidly within the first 20 min, with a productivity rate of 2.19 g PHA/L-h. The results in this study indicate that PHA production by using activated sludge is a promising alternative to a typical pure culture approach.

Bacteria, Aerobic↗

Effects of anaerobic/aerobic incubation and storage temperature on preservation and deodorization of kitchen garbage.

To develop a garbage recycling system for the purpose of the production of lactic acid (LA) to use as raw material for producing biodegradable plastics, the preservation and deodorization of garbage during storage are very important. Anaerobic incubation (i.e., storage) was prove to be more suitable than aerobic incubation during the garbage storage in terms of concentration of LA and soluble sugar, pH value, viable bacteria counts and offensive odour substances. This difference is due to a fact that the growth of putrefactive bacteria such as coliforms and Clostridium spp. appeared to be inhibited by anaerobic fermentation during the storage, because the fermentation caused a drop of garbage pH and generated inhibitory substances, i.e., bacteriocins. Under anaerobic condition, LA concentration in the stored garbage was found to be higher in the order: 37 > 25 > 50 > 5 degrees C, and the concentration of sugar accumulated during the 50 degrees C-storage was the highest. Among the conditions employed, the optimum condition for the storage of kitchen garbage was anaerobic at 5 degrees C.

Bacteria, Aerobic↗

Effects of intermittent addition of cellulase for production of L-lactic acid from wastewater sludge by simultaneous saccharification and fermentation.

An attempt was made to create L-lactic acid, a precursor of poly-lactic acid, which is a biodegradable plastic, from wastewater sludge from the paper-manufacturing industry. The sludge contained a high percentage of cellulose and needed to be hydrolyzed to glucose by the action of the cellulase before being treating with lactic acid bacteria. Therefore, a method involving simultaneous saccharification and fermentation (SSF) was carried out. The optimum pH of the SSF for production of the lactic acid by the newly isolated lactic acid bacterium with a high selectively of L-lactic acid was found out to be around pH = 5.0, and the optimum temperature to be approximately 40 degrees C. On the basis of the measurement of the cell density changes in the lactic acid bacteria, it was ascertained that the bacterial activity could continue at a high level for a relatively long period of time, and that the L-lactic acid productivity was diminished by the rapid deactivation of the cellulase. With the intermittent addition of cellulase once daily for the sake of compensating for the cellulase deactivation, the L-lactic acid attained a maximum concentration of 16.9 g/L, i.e., a 72.2% yield based on the potential glucose contained in the sludge under optimum pH and temperature conditions.

Biodegradation, Environmental↗

Bioconversion of kitchen garbage to lactic acid by two wild strains of Lactobacillus species.

To enhance lactic acid (LA) production from kitchen garbage, which is a raw material for biodegradable plastics production, the application of high-performance lactic acid bacteria (LAB) as inocula was investigated. Two wild strains of Lactobacillus species, designated as TH165 and TD175, were isolated and screened from kitchen garbage. Strain TH165 was capable of hydrolyzing starch to produce LA; 49.5% of starch was broken down in fermentation medium containing 8.52 g/L of soluble starch, and 4.01 g/L of LA was produced after 24 h fermentation at 37 degrees C without pH control. Strain TD175 could produce 16.06 g/L of LA, 66.9% higher than that of Lactobacillus bulgaricus ACCC11058 in fermentation medium containing 2.0% glucose at 30 degrees C without pH control. Furthermore, coinoculation of strains TH165 and TD175 enhanced the LA production, resulting in 33.80 g/L of LA concentration and 0.46 g/g (DW) of LA yield from nonautoclaved kitchen garbage after 72 h fermentation with pH maintained at 5.5-6.0, values 36.9% higher than those of the fermentation without inoculum (control). This study shows that enhancement of LA production from kitchen garbage can be realized by using high-performance LAB. This recycling system is conducive to clear away pollutants and to reduce cost of LA production.

Biodegradation, Environmental↗

Restructuring upstream bioprocessing: technological and economical aspects for production of a generic microbial feedstock from wheat.

Restructuring and optimization of the conventional fermentation industry for fuel and chemical production is necessary to replace petrochemical production routes. Guided by this concept, a novel biorefinery process has been developed as an alternative to conventional upstream processing routes, leading to the production of a generic fermentation feedstock from wheat. The robustness of Aspergillus awamori as enzyme producer is exploited in a continuous fungal fermentation on whole wheat flour. Vital gluten is extracted as an added-value byproduct by the conventional Martin process from a fraction of the overall wheat used. Enzymatic hydrolysis of gluten-free flour by the enzyme complex produced by A. awamori during fermentation produces a liquid stream rich in glucose (320 g/L). Autolysis of fungal cells produces a micronutrient-rich solution similar to yeast extract (1.6 g/L nitrogen, 0.5 g/L phosphorus). The case-specific combination of these two liquid streams can provide a nutrient-complete fermentation medium for a spectrum of microbial bioconversions for the production of such chemicals as organic acids, amino acids, bioethanol, glycerol, solvents, and microbial biodegradable plastics. Preliminary economic analysis has shown that the operating cost required to produce the feedstock is dependent on the plant capacity, cereal market price, presence and market value of added-value byproducts, labor costs, and mode of processing (batch or continuous). Integration of this process in an existing fermentation plant could lead to the production of a generic feedstock at an operating cost lower than the market price of glucose syrup (90% to 99% glucose) in the EU, provided that the plant capacity exceeds 410 m(3)/day. Further process improvements are also suggested.

Aspergillus↗

Production of microbial polyesters: fermentation and downstream processes.

Poly(3-hydroxyalkanoates) (PHAs) constitute a large and versatile family of polyesters produced by various bacteria. PHAs are receiving considerable attention because of their potential as renewable and biodegradable plastics, and as a source of chiral synthons since the monomers are chiral. Industrial PHA production processes have been developed for poly(3-hydroxybutyrate) (poly(3HB)) and poly(3-hydroxybutyrate-co-3-valerate) (poly(3HB-co-3HV). More than 100 other poly(3HAMCL)s, characterized by monomers of medium chain length, have been identified in the past two decades. These monomers typically contain 6-14 carbon atoms, are usually linked via-3-hydroxy ester linkages, but can occasionally also exhibit 2-, 4-, 5-, or 6-hydroxy ester linkages. Such polyesters are collectively referred to as medium chain length PHAs poly(3HAMCL)s. The vast majority of these interesting biopolyesters have been studied and produced only on the laboratory scale. However, there have been several attempts to develop pilot scale processes, and these provide some insight into the production economics of poly(3HAMCL)s other than poly(3HB) and poly(3HB-co-3HV). These processes utilize diverse fermentation strategies to control the monomer composition of the polymer, enabling the tailoring of polymer material properties to some extent. The best studied of these is poly(3-hydroxyoctanoate) (poly(3HO)), which contains about 90% 3-hydroxyoctanoate. This biopolyester has been produced on the pilot scale and is now being used in several experimental applications.

Chemical Engineering↗

[Memory effect of resorbable polymers].

BACKGROUND: The present study analyzed the memory effect of resorbable polymers. Depending on temperature, this effect describes the ability of different materials to "remember" their original form after mechanical deformation. Resorbable polymers serve as materials to stabilize and fix bone fractures. Compared to metal transplants, resorbable polymers are able to undergo thermoplastic deformation. The precise adaptation of the transplant to the surrounding bone guarantees an exact anatomical reconstruction. However, during normal applications, it was observed that these biodegradable plastic materials tend to revert to their original form at body temperature. This "memory effect" could result in negative consequences for the medical treatment. METHODS: By the process of compression molding, geometrically formed specimens (lattice, rod, plate) consisting of different polyglycolides and polylactides were prepared. After warming up the specimens to 50 degrees C (water bath) they were deformed into definite angles. Following this procedure, the specimens were put in a water bath at 37 degrees C to mimic the adaptation of the transplant at body temperature. The retroflexion of the material (memory effect) was measured using an XY-measuring desk. RESULTS: The present study clearly reveals that highly deformed specimens react with stronger retroflexions. In addition, the results indicate that the memory effect depends on geometrical design as well as on chemical composition. All tested polymers showed a strong initial memory effect that decreased with time.

Absorbable Implants↗

The Pseudomonas aeruginosa RhlA enzyme is involved in rhamnolipid and polyhydroxyalkanoate production.

Pseudomonas aeruginosa produces the biosurfactant rhamnolipid, which has several potential biotechnological applications. The synthesis of this surfactant is catalyzed by rhamnosyltransferase 1, composed of the proteins RhlA and RhlB. Here we report that RhlA plays a role not only in surfactant synthesis, but also in the production of polyhydroxyalkanoates, polymers that can be used for the synthesis of biodegradable plastics.

Bacterial Proteins↗

Synthesis of PHAs from waster under various C:N ratios.

Polyhydroxyalkanoates (PHAs) production was carried out under various C:N ratios. A ratio of 100 resulted best polymer yield. C-source was an important factor in synthesis. For example, as the ratio of valeric acid (C5) to butyric acid (C4) in N-free medium was increased, the mole fraction of HV in the copolymer increased. When soy waste was used as a C-source a copolymer, a high HV mole fraction (HB:HV, 75:25) was produced while when malt waste was used, a much lower HV mole fraction (HB:HV, 90:10) was generated. It was concluded that activated sludge bacteria could be induced to produce PHAs using food wastes as C-sources and this could be the basis for production of biodegradable plastics.

Alcaligenes↗