Hong Chua

Bio-accumulation of environmental residues of rare earth elements in aquatic flora Eichhornia crassipes (Mart.) Solms in Guangdong Province of China

Scattering and bio-accumulation of rare earth elements (REEs), including the inner transition series, in the aquatic environment in southern China have resulted from increased industrial and agricultural applications. Environmental residues of REEs entered into aquatic flora, namely Eichhornia crassipes, commonly known as water hyacinth, via the root system in contaminated substrate water and could distribute to various parts of the plant. REEs could also bio-accumulate and concentrate in the leaves at a concentration ratio of approx. 3 times regardless of initial REE concentration in the substrate water. REEs could also enter into the plant via the leaves that are exposed to atmospheric contaminants. While officially permitted residual concentrations of mixed REE nitrates in foodstuffs or animal feedstocks are not available, high REE concentrations in the substrate water in which water hyacinth grew could possibly enter the human food chain and lead to adverse public health problems.

Biosorption of heavy metals by bacteria isolated from activated sludge

Twelve aerobic bacteria from activated sludge were isolated and identified. These included both Gram-positive (e.g., Bacillus) and Gram-negative (e.g., Pseudomonas) bacteria. The biosorption capacity of these strains for three different heavy metals (copper, nickel, and lead) was determined at pH 5.0 and initial metal concentration of 100 mg/L. Among these 12 isolates, Pseudomonas pseudoalcaligenes was selected for further investigation owing to its high metal biosorption capacity. The lead and copper biosorption of this strain followed the Langmuir isotherm model quite well with maximum biosorption capacity (qmax) reaching 271.7mg of Pb2+/g of dry cell and 46.8 mg of Cu2+/g of dry cell at pH 5.0. Study of the effect of pH on lead and copper removal indicated that the metal biosorption increased with increasing pH from 2.0 to 7.0. A mutual inhibitory effect was observed in the lead-copper system because the presence of either ion affected the sorption capacity of the other. Unequal inhibitions were observed in all the nickel binary systems. The increasing order of affinity of the three metals toward P. pseudoalcaligenes was Ni<Cu<Pb. The metal biosorptive potential of these isolates, especially P. pseudoalcaligenes, may have possible applications in the removal and recovery of metals from industrial effluents.

Conversion of Food Industrial Wastes into Bioplastics

The usage of plastics in packaging and disposable products, and the generation of plastic waste, have been increasing drastically. 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. In the authors’ laboratories, various carbohydrates in the growth media, including sucrose, lactic acid, butyric acid, valeric acid, and various combinations of butyric and valeric acids, were utilized as the carbon (c) sources for the production of bioplastics byAlcaligenes eutrophus. As the first step in pursuit of eventual usage of industrial food wastewater as nutrients for microorganisms to synthesize bioplastics, the authors investigated the usage of malt wastes from a beer brewery plant as the C sources for the production of bioplastics by microorganisms. Specific polymer production yield by A. Latus DSM 1124 increased to 70% polymer/cell (g/g) and 32g/L cell dry wt, using malt wastes as the C source. The results of these experiments indicated that, with the use of different types of food wastes as the C source, different polyhydroxyal-kanoate copolymers could be produced with distinct polymer properties.

Accumulation of biopolymers in activated sludge biomass.

In this study, activated sludge bacteria from a conventional wastewater treatment process were induced to accumulate polyhydroxyalkanoates (PHAs) under different carbon-nitrogen (C:N) ratios. As the C:N ratio increased from 20 to 140, specific polymer yield increased to a maximum of 0.38 g of polymer/g of dry cell mass while specific growth yield decreased. The highest overall polymer production yield of 0.11 g of polymer/g of carbonaceous substrate consumed was achieved using a C:N ratio of 100. Moreover, the composition of polymer accumulated was dependent on the valeric acid content in the feed. Copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] was produced in the presence of valeric acid. The 3-hydroxyvalerate (3HV) mole fraction in the copolymer was linearly related to valeric content in the feed, which reached a maximum of 54% when valeric acid was used as sole carbon source. When the 3HV U in the polymer increased from 0-54 mol%, the melting temperature decreased from 178 degrees to 99 degrees C. Thus, the composition, and hence the mechanical properties, of the copolymer produced from activated sludge can be controlled by adjusting the mole fraction of valeric acid in the feed medium.

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.

Coupling of Waste Water Treatment with Storage Polymer Production

Storage polymers in bacterial cells can be extracted and used as biodegradable thermoplastics. However, widespread applications have been limited by high production costs. In this study, activated sludge bacteria in a conventional waste water treatment system were induced, by controlling the carbon-nitrogen (C:N) ratio in the reactor liquor, to accumulate storage polymers. Specific polymer yield increased to a maximum of 0.374 g polymer/g cell when the C:N ratio was increased from from 24 to 144, whereas specific growth yield decreased with increasing C:N ratio. An optimum C:N ratio of 96 provided the highest overall polymer production yield of 0.093 g of polymer/g of carbonaceous substrate consumed, without significantly affecting the organic treatment efficiency in the waste water treatment system.

Construction of recombinant Bacillus subtilis strains for polyhydroxyalkanoates synthesis

Abstract Plastic wastes are considered to be a worldwide environmental problem and demand for biodegradable plastics has become a highly visible issue. One of the most important characteristics of microbial polyesters is that they are thermoplastic with environmentally degradable properties. In this experiment, pSG703/ pha RBC and pSG703/ pha PQRBC were cloned and transformed into φ105 prophage-based Bacillus subtilis . Construction of recombinant Bacillus was only partially successful. The pha PQRBC genes from B. megaterium were cloned into B. subtilis 1A304 (φ105 MU331). The pha genes is stable in the absence of selective pressure because the prophage is covalently inserted, in a single copy, into the host chromosome. The B. subtills 1A304 (φ105 MU331) carrying the pha genes ( pha PQRBC) was subjected to fermentation and showed PHA accumulation, which was the first report of the expression of the pha genes of B. megaterium in B. subtilis . The cells was subjected for FT-IR and GC analysis and the product was identified to be a PHB homopolymer. The recombinant B. subtilis was expressed with heat shock process also had the PHA accumulation but with the yield lower than the same strain without heat shock that might be due to the RNA polymerase bound to phage promoter and native promoters of pha PQ gene, respectively, and then was run along in the opposite direction during the transcription. The results also showed that the recombinant B. subtilis could utilize the malt waste in the medium as a carbon source better than that of glucose and thus could substantially lowered the cost of production of PHA.

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 outperfomed 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.

Biosorption and Desorption of Copper (II) Ions by Bacillus sp

Batch biosorption experiments were conducted to investigate the removal of Cu2+ ions from aqueous solutions by a series of bacterial strains isolated from a local activated sludge process. The characteristics of 12 isolates were identified and examined for their ability to bind Cu2+ ions from aqueous solution. Among the isolates, two species exhibited biosorption capacity >40 mg of Cu/g of dry cell. Isotherms for the biosorption of copper on bacterial cells were developed and compared, and the equilibrium data fitted well to the Langmuir and Freundlich isotherm models. The biosorption of copper increased significantly with increasing pH from 2.0 to 6.0 regardless of the species. More than 90% of copper sorbed on the cells of Bacillus sp. could be recovered by washing with 0.1 M HNO3 for 5 min. The performance of two different desorption processes was also tested and compared. The results show that five biosorption and desorption cycles are a better operation process than five successive biosorptions followed by one desorption to remove and recover copper from aqueous solution. The biosorbent could be used for at least five biosorptions and desorption cycles without loss of copper removal capacity. It can be concluded that the activated sludge or sludge-isolated bacteria could be a potential biosorbent for copper removal.

Removal and recovery of copper (II) ions by bacterial biosorption

Studies were conducted toinvestigate the removal and recovery of copper (II) ions from aqueous solutions by Micrococcus sp., which was isolated from a local activated sludge process. The equilibrium of copper biosorption followed the Langmuir isotherm model very well with a maximum biosorption capacity (qmax) of 36.5 mg of Cu2+/gofdry cell at pH 5.0 and 52.1 mg of Cu2+/g of dry cell at pH 6.0. Cells harvested at exponential growth phase and stationary phase showed similar biosorption characteristics for copper, Copper uptake by cells was negligible at pH 2.0 and then increased rapidly with increasing pH un til 6.0. In multim etal systems, Micrococcus sp. exhibited a preferential biosorption order: Cu−Pb>Ni−Zn. There is virtually no interference with copper uptake by Micrococcus sp. from solutions bearing high concentrations of Cl−, SO42−, and NO3/− (0–500 mg/L). Sulfuric acid (0.05 M) was the most efficient desorption medium, recovering >90% of the initial copper sorbed. The copper capacity of Micrococcus sp. remained unchanged after five successive sorption and desorption cycles. Immobilization of Micrococcus sp. in 2% calcium alginate and 10% polyacrylamide gel beads increased copper uptake by 61%. Biomass of Micrococcus sp. may be applicable to the development of potentially cost-effective biosorbent for removing and recovering copper from effluents.