Computational Identification of Bacterial Communities

Stable bacterial polymorphism on a single limiting resource may appear if between the evolved strains metabolic interactions take place that allow the exchange of essential nutrients [8]. Towards an attempt to predict the possible outcome of longrunning evolution experiments, a network based on the metabolic capabilities of homogeneous populations of every single gene knockout strain (nodes) of the bacterium E. coli is reconstructed. Potential metabolic interactions (edges) are allowed only between strains of different metabolic capabilities. Bacterial communities are determined by finding cliques in this network. Growth of the emerged hypothetical bacterial communities is simulated by extending the metabolic flux balance analysis model of Varma et al [2] to embody heterogeneous cell population growth in a mutual environment. Results from aerobic growth on 10 different carbon sources are presented. The upper bounds of the diversity that can emerge from single-cloned populations of E. coli such as the number of strains that appears to metabolically differ from most strains (highly connected nodes), the maximum clique size as well as the number of all the possible communities are determined. Certain single gene deletions are identified to consistently participate in our hypothetical bacterial communities under most environmental conditions implying a pattern of growth-condition- invariant strains with similar metabolic effects. Moreover, evaluation of all the hypothetical bacterial communities under growth on pyruvate reveals heterogeneous populations that can exhibit superior growth performance when compared to the performance of the homogeneous wild-type population.

Denitrification of Wastewater Containing High Nitrate Using a Bioreactor System Packed by Microbial Cellulose

A Laboratory-scale packed bed reactor with microbial cellulose as the biofilm carrier was used to investigate the denitrification of high-strength nitrate wastewater with specific emphasis on the effect the nitrogen loading rate and hydraulic retention time. Ethanol was added as a carbon source for denitrification. As a result of this investigation, it was found that up to 500 mg/l feed nitrate concentration the present system is able to produce an effluent with nitrate content below 10 ppm at 3 h hydraulic retention time. The highest observed denitrification rate was 4.57 kg NO3-N/ (m3 .d) at a nitrate load of 5.64 kg NO3- N/(m3 .d), and removal efficiencies higher than 90% were obtained for loads up to 4.2 kg NO3-N/(m3 .d). A mass relation between COD consumed and NO3-N removed around 2.82 was observed. This continuous-flow bioreactor proved an efficient denitrification system with a relatively low retention time.

Application of Genetic Engineering for Chromium Removal from Industrial Wastewater

The treatment of the industrial wastewater can be particularly difficult in the presence of toxic compounds. Excessive concentration of Chromium in soluble form is toxic to a wide variety of living organisms. Biological removal of heavy metals using natural and genetically engineered microorganisms has aroused great interest because of its lower impact on the environment. Ralston metallidurans, formerly known as Alcaligenes eutrophus is a LProteobacterium colonizing industrial wastewater with a high content of heavy metals. Tris-buffered mineral salt medium was used for growing Alcaligenes eutrophus AE104 (pEBZ141). The cells were cultivated for 18 h at 30 oC in Tris-buffered mineral salt medium containing 3 mM disodium sulphate and 46 mM sodium gluconate as the carbon source. The cells were harvested by centrifugation, washed, and suspended in 10 mM Tris HCl, pH 7.0, containing 46 mM sodium gluconate, and 5 mM Chromium. Interaction among induction of chr resistance determinant, and chromate reduction have been demonstrated. Results of this study show that the above bacteria can be very useful for bioremediation of chromium from industrial wastewater.

Carbon Sources Utilization Profiles of Thermophilic Phytase Producing Bacteria Isolated from Hot-spring in Malaysia

Phytases (myo-inositol hexakisphosphate phosphohydrolases; EC 3.1.3.8) catalyze the hydrolysis of phytic acid (myoinositol hexakisphosphate) to the mono-, di-, tri-, tetra-, and pentaphosphates of myo-inositol and inorganic phosphate. Therrmophilic bacteria isolated from water sampled from hot spring. About 120 isolates of bacteria were successfully isolated form hot spring water sample and tested for extracellular phytase producing. After 5 passages of the screening on the PSM media, 4 isolates were found stable in producing phytase enzyme. The 16s RDNA sequencing for identification of bacteria using molecular technique revealed that all isolates those positive in phytase producing are belong to Geobacillus spp. And Anoxybacillus spp. Anoxybacillus rupiensis UniSZA-7 were identified for their carbon source utilization using Phenotype Microarray Plate of Biolog and found they utilize several kind of carbon source provided.

Using Artificial Neural Network and Leudeking-Piret Model in the Kinetic Modeling of Microbial Production of Poly-β- Hydroxybutyrate

Poly-β-hydroxybutyrate (PHB) is one of the most famous biopolymers that has various applications in production of biodegradable carriers. The most important strategy for enhancing efficiency in production process and reducing the price of PHB, is the accurate expression of kinetic model of products formation and parameters that are effective on it, such as Dry Cell Weight (DCW) and substrate consumption. Considering the high capabilities of artificial neural networks in modeling and simulation of non-linear systems such as biological and chemical industries that mainly are multivariable systems, kinetic modeling of microbial production of PHB that is a complex and non-linear biological process, the three layers perceptron neural network model was used in this study. Artificial neural network educates itself and finds the hidden laws behind the data with mapping based on experimental data, of dry cell weight, substrate concentration as input and PHB concentration as output. For training the network, a series of experimental data for PHB production from Hydrogenophaga Pseudoflava by glucose carbon source was used. After training the network, two other experimental data sets that have not intervened in the network education, including dry cell concentration and substrate concentration were applied as inputs to the network, and PHB concentration was predicted by the network. Comparison of predicted data by network and experimental data, indicated a high precision predicted for both fructose and whey carbon sources. Also in present study for better understanding of the ability of neural network in modeling of biological processes, microbial production kinetic of PHB by Leudeking-Piret experimental equation was modeled. The Observed result indicated an accurate prediction of PHB concentration by artificial neural network higher than Leudeking- Piret model.

Sophorolipids Production by Candida Bombicola using Synthetic Dairy Wastewater

Sophorolipids (SLs) production by the yeast Candida bombicola was studied in batch shake flasks using synthetic dairy wastewaters (SDWW) with or without any added external carbon and nitrogen sources. A maximum SLs production of 38.76 g/l was observed with the SDWW supplemented with low cost substrate of sugarcane molasses at 50 g/l and soybean oil at 50 g/l. When the SDWW was supplemented with more costly glucose, yeast extract, urea and soybean oil, the production, however, got lowered to only 29.49 g/l, but with a maximum biomass production of 17.38 g/l together with a complete utilization of the carbon sources.

A Green Chemical Technique for the Synthesis of Magnetic Nanoparticles by Magnetotactic Bacteria

Bacterial magnetic nanoparticles have great useful potential in biotechnological and biomedical applications. In this study, a liquid growth medium was modified for cultivation a fastidious magnetotactic bacterium that has been isolated from Anzali lagoon, Iran in our previous research. These modifications include change in vitamin, mineral, carbon sources and etcetera. In our experience, the serum bottles and designed air-tight laboratory bottles were used to create microaerobic conditions in order to development of a method for scale-up experiment. This information may serve as a guide to green chemistry based biological protocols for the synthesis of magnetic nanoparticles with control over the chemical composition, morphology and size.

Microbial Oil Production by Isolated Oleaginous Yeast Torulaspora globosa YU5/2

Microbial oil was produced by soil isolated oleaginous yeast YU5/2 in flask-batch fermentation. The yeast was identified by molecular genetics technique based on sequence analysis of the variable D1/D2 domain of the large subunit (26S) ribosomal DNA and it was identified as Torulaspora globosa. T. globosa YU5/2 supported maximum values of 0.520 g/L/d, 0.472 g lipid/g cells, 4.16 g/L, and 0.156 g/L/d for volumetric lipid production rate, and specific yield of lipid, lipid concentration, and specific rate of lipid production respectively, when culture was performed in nitrogen-limiting medium supplemented with 80g/L glucose. Among the carbon sources tested, maximum cell yield coefficient (YX/S, g/L), maximum specific yield of lipid (YP/X, g lipid/g cells) and volumetric lipid production rate (QP, g/L/d) were found of 0.728, 0.237, and 0.619, respectively, using sweet potato tubers hydrolysates as carbon source.

Regulatory Effects of Carbon Sources on Tabtoxin Production (A β-lactam Phytotoxin of Pseudomonas syringae pv. tabaci)

The effects of divers carbon substrates were investigated for the tabtoxin production of an isolated pathogenic Pseudomonas syringae pv. tabaci, the causal agent of wildfire of tobacco and are discussed in relation to the bacterium growth. The isolated organism was grown in batch culture on Woolley's medium (28°C, 200 rpm, during 5 days). The growth has been measured by the optical density (OD) at 620 nm and the tabtoxin production quantified by Escherichia coli (K-12) bioassay technique. The growth and the tabtoxin production were both influenced by the substrates (sugars, amino acids, organic acids) used, each, as a sole carbon source and as a supplement for the same amino acids. The most significant quantities of tabtoxin were obtained in presence of some amino acids used as sole carbon source and/or as supplement.

Enhance Halorespiration in Rhodopseudomonas palustris with Cytochrome P450cam System from Pseudomonas putida

To decompose organochlorides by bioremediation, co-culture biohydrogen producer and dehalogenation microorganisms is a useful method. In this study, we combined these two characteristics from a biohydrogen producer, Rhodopseudomonas palustris, and a dehalogenation microorganism, Pseudomonas putida, to enchance halorespiration in R. palustris. The genes encoding cytochrome P450cam system (camC, camA, and camB) from P. putida were expressed in R. palustris with designated expression plasmid. All tested strains were cultured to log phase then presented pentachloroethane (PCA) in media. The vector control strain could degrade PCA about 78% after 16 hours, however, the cytochrome P450cam system expressed strain, CGA-camCAB, could completely degrade PCA in 12 hours. While taking chlorinated aromatic, 3-chlorobenzoate, as sole carbon source or present benzoate as co-substrate, CGA-camCAB presented faster growth rate than vector control strain.

Growth Behaviors, Thermostable Direct Hemolysin Secretion and Fatty Acid Profiles of Acid-adapted and Non-adapted Vibrio parahaemolyticus

Three strains of Vibrio parahaemolyticus (690, BCRC 13023 and BCRC 13025) implicated in food poisoning outbreaks in Taiwan were subjected to acid adaptation at pH 5.5 for 90 min. The growth behaviors of acid-adapted and non-adapted V. parahaemolyticus in the media supplemented with various nitrogen and carbon sources were investigated. The effects of acid adaptation on the thermostable direct hemolysin (TDH) secretion and fatty acid profiles of V. parahaemolyticus were also examined. Results showed that acid-adapted and non-adapted V. parahaemolyticus 690, BCRC 13023 and BCRC 13025 grew similarly in TSB-3% NaCl and basal media supplemented with various carbon and nitrogen sources during incubation period. Higher TDH secretion was noted with V. parahaemolyticus 690 among the three strains. However, acid-adapted strains produced less amounts of TDH than non-adapted strains when they were grown in TSB-3% NaCl. Additionally, acid adaptation increased the ratio of SFA/USFA in cells of V. parahaemolyticus strains.

Process Optimization for Enhanced Production of Cell Biomass and Metabolites of Fluorescent Pseudomonad R81

The fluorescent pseudomonad strain R81 is a root colonizing rhizobacteria which promotes the growth of many plants by various mechanisms. Its broth containing siderophore (ironchelating compound) and 2,4- diacetyl phloroglucinol (DAPG) is used for preparing bioinoculant formulations for agronomical applications. Glycerol was found to be the best carbon source for improved biomass production. Splitting of nitrogen source to NH4Cl and urea had a stabilizing effect on pH during batch cultivation. Ltryptophan at 0.5 % in the medium increased the siderophore production to 850 mg/l. During batch cultivation of the strain in a bioreactor, a maximum of 4 g/l of dry cell mass, 1.8 g/l of siderophore and 20 mg/l of DAPG was achieved when glycerol was 15 g/l and C/N ratio was maintained at 12.5. In case of intermittent feeding of fresh medium during fed-batch cultivation, the dry cell mass was increased to 25 g/l with improved production of DAPG to 70 mg/l.

Optimal Conditions for Carotenoid Production and Antioxidation Characteristics by Rhodotorula rubra

This study aims to screen out and to optimize the major nutrients for maximum carotenoid production and antioxidation characteristics by Rhodotorula rubra. It was found that supplementary of 10 g/l glucose as carbon source, 1 g/l ammonium sulfate as nitrogen source and 1 g/l yeast extract as growth factor in the medium provided the better yield of carotenoid content of 30.39 μg/g cell dry weight the amount of antioxidation of Rhodotorula rubra by DPPH, ABTS and MDA method were 1.463%, 34.21% and 34.09 μmol/l, respectively.

Starch Based Biofilms for Green Packaging

This current research focused on development of degradable starch based packaging film with enhanced mechanical properties. A series of low density polyethylene (LDPE)/tapioca starch compounds with various tapioca starch contents were prepared by twin screw extrusion with the addition of maleic anhydride grafted polyethylene as compatibilizer. Palm cooking oil was used as processing aid to ease the blown film process, thus, degradable film can be processed via conventional blown film machine. Studies on their characteristics, mechanical properties and biodegradation were carried out by Fourier Transform Infrared (FTIR) spectroscopy and optical properties, tensile test and exposure to fungi environment respectively. The presence of high starch contents had an adverse effect on the tensile properties of LDPE/tapioca starch blends. However, the addition of compatibilizer to the blends improved the interfacial adhesion between the two materials, hence, improved the tensile properties of the films. High content of starch amount also was found to increase the rate of biodegradability of LDPE/tapioca starch films. It can be proved by exposure of the film to fungi environment. A growth of microbes colony can be seen on the surface of LDPE/tapioca starch film indicates that the granular starch present on the surface of the polymer film is attacked by microorganisms, until most of it is assimilated as a carbon source.

Fabrication of Carbon Doped TiO2 Nanotubes via In-situ Anodization of Ti-foil in Acidic Medium

Highly ordered TiO2 nanotube (TNT) arrays were fabricated onto a pre-treated titanium foil by anodic oxidation with a voltage of 20V in phosphoric acid/sodium fluoride electrolyte. A pretreatment of titanium foil involved washing with acetone, isopropanol, ethanol and deionized water. Carbon doped TiO2 nanotubes (C-TNT) was fabricated 'in-situ' with the same method in the presence of polyvinyl alcohol and urea as carbon sources. The affects of polyvinyl alcohol concentration and oxidation time on the composition, morphology and structure of the C-TN were studied by FE-SEM, EDX and XRD techniques. FESEM images of the nanotubes showed uniform arrays of C-TNTs. The density and microstructures of the nanotubes were greatly affected by the content of PVA. The introduction of the polyvinyl alcohol into the electrolyte increases the amount of C content inside TiO2 nanotube arrays uniformly. The influence of carbon content on the photo-current of C-TNT was investigated and the I-V profiles of the nanotubes were established. The preliminary results indicated that the 'in-situ' doping technique produced a superior quality nanotubes compared to post doping techniques.

Microalgal Lipid Production by Microalgae Chlorella sp. KKU-S2

The objective of this work is to produce heterotrophic microalgal lipid in flask-batch fermentation. Chlorella sp. KKU-S2 supported maximum values of 0.374 g/L/d, 0.478 g lipid/g cells, and 0.112 g/L/d for volumetric lipid production rate, and specific yield of lipid, and specific rate of lipid production, respectively when culture was performed on BG-11 medium supplemented with 50g/L glucose. Among the carbon sources tested, maximum cell yield coefficient (YX/S, g/L), maximum specific yield of lipid (YP/X, g lipid/g cells) and volumetric lipid production rate (QP, g/L/d) were found of 0.728, 0.237, and 0.619, respectively, using sugarcane molasses as carbon source. The main components of fatty acid from extracted lipid were palmitic acid, stearic acid, oleic acid and linoleic acid which similar to vegetable oils and suitable for biodiesel production.

The Composition of Rice Bran Hydrolysate and Its Possibility to Use in the Ethanol Production by Zymomonas mobilis Biofilm

Rice bran has been abandoned as agricultural waste for million tonnes per year in Thailand, therefore they have been proposed to be utilized as a rich carbon source in the production of bioethanol. Many toxic compounds are possibly released during the pretreatment of rice bran prior the fermentation process. This study aims to analyze on the availability of toxic compounds and the amount of glucose obtained from 2 different pretreatments using sulfuric acid and mixed cellulase enzymes (without and with delignification/ activated charcoal). The concentration of furfural, 5- hydroxymethyl furfural (5-HMF), levulinic acid, vanillin, syringaldehyde and4-hydroxybenzaldehyde (4-HB) and the percent acetic acid were found to be 0.0517 ± 0.049 mg/L, 0.032 ± 0.06 mg/L, 21074 ± 1685.62 mg/L, 126.265 ± 6.005 mg/L, 2.89 ± 0.30 mg/L, 0.37 ± 0.031mg/L and 0.72% under the pretreatment process without delignification/ activated charcoal treatment and 384.47 ± 99.02 g/L, 0.068 mg/L, 142107.62 ± 8664.6 mg/L, 0.19 mg/L, 5.43 ± 3.29 mg/L, 4.80 ± 0.76 mg/L and 0.254% under the pretreatment process with delignification/ activated charcoal treatment respectively. The presence of high concentration of acetic acid was found to impede the growth of Zymomonas mobilis strain TISTR 551 despite the present of high concentration of levulinic acid. Z. mobilis strain TISTR 551 was found to produce 8.96 ± 4.06 g/L of ethanol under 4 days fementation period in biofilm stage in which represented 40% theoretical yield.

Effect of Fermentation Time on Xanthan Gum Production from Sugar Beet Molasses

Xanthan gum is a microbial polysaccharide of great commercial significance. The purpose of this study was to select the optimum fermentation time for xanthan gum production by Xanthomonas campestris (NRRL-B-1459) using 10% sugar beet molasses as a carbon source. The pre-heating of sugar beet molasses and the supplementation of the medium were investigated in order to improve xanthan gum production. Maximum xanthan gum production in fermentation media (9.02 g/l) was observed after 4 days shaking incubation at 25°C and 240 rpm agitation speed. A solution of 10% sucrose was used as a control medium. Results indicated that the optimum period for xanthan gum production in this condition was 4 days.