Bioflocculant produced by Chlamydomonas reinhardtii

Bioflocculants of Chlamydomonas reinhardtii were investigated under axenic conditions. C. reinhardtii was found to produce significant amounts of bioflocculants. Flocculating activity by C. reinhardtii began in the linear phase of growth and continued until the end of the stationary phase. The highest flocculating efficiency of the culture broth was 97.06%. The purified C. reinhardtii bioflocculant was composed of 42.1% (w/w) proteins, 48.3% carbohydrates, 8.7% lipids, and 0.01% nucleic acid. The optimum condition for bioflocculant production of C. reinhardtii was as follows: under temperature of 15°C to 25°C, pH 6–10 and illumination of 40–60 μmol photons m^?2 s^?1. The bioflocculants produced by C. reinhardtii showed maximum activity in pH ranges from 2 to 10. The flocculating activity was significantly enhanced by the addition of CaCl₂ as a co-flocculant at an optimal concentration of 4.5 mM.     

Characterization of bioflocculants from biologically aerated filter backwashed sludge and its application in dying wastewater treatment

In this study, the feasibility of bioflocculant extraction from backwashing sludge to reduce its production costs was investigated. Results showed that ultrasound and base treatment could significantly enhance bioflocculant extraction efficiency, however, flocculating activity was affected. It was observed that bioflocculants extracted from sludge of pH 11.0 had no flocculating activity. In contrast, bioflocculants extracted from sludge of pH 5.0, named as M-1, had good flocculating activity. To further study the flocculating activity of M-1, factors such as bioflocculant dosage, temperature and pH of the reaction solution were tested. The optimal conditions were 6.0 mg/l bioflocculant dosage and pH 5.0, at a temperature of 10 °C. Under these conditions, the flocculating rate of kaolin clay was 92.67%. The effectiveness of such bioflocculants in the decolorization of synthetically dyed wastewater was then examined. In flocculating methylene blue and fast blue in aqueous solutions, decolorization efficiency levels were 82.9% and 77.8%, respectively.     

Bioconversion of lignocellulose and simultaneous production of cellulase,ligninase and bioflocculants by Alcaligenes faecalis-X3

Lignocelluloses have been used as carbon sources for bioflocculant production. However, the low bioconversion efficiency of lignocellulose to bioflocculants is a major challenge. In this study, a lignocellulolytic strain of Alcaligenes faecalis-X3 was cultivated in ramie bio-degumming wastewater. Optimal production of ligninase, cellulase and bioflocculants (MBF-X3) was evaluated. The highest activity of MBF-X3 under the optimal conditions of pH 6.0 at 48 h of fermentation was 95.44%, with the maximum production of ligninase and cellulase (0.27 and 0.12 U/mL, respectively). The crude ligninase and cellulase had optimum activities at pH 5.0 and 40 °C and pH 6.0 and 50 °C, respectively. The cellulase activity was increased by Mn²⁺, Ca²⁺, Zn²⁺, and Mg²⁺ at 1 mM. The ligninase activity was significantly enhanced in the presence of Zn²⁺ at 10 mM. The flocculating activity of MBF-X3 was not changed by the addition of any metal cation. The results demonstrated that A. faecalis possesses an excellent enzyme system for the efficient bioconversion of lignocellulose into MBF-X3. Additionally, MBF-X3 has a high flocculating efficiency of Disperse Blue-2BLN (85.7%) at a dose of 1.0 g/L.     

Characterization of metal-binding bioflocculants produced by the cyanobacterial component of mixed microbial mats.

Mixed-species microbial mats that were dominated by the cyanobacterium Oscillatoria sp. and contained heterotrophic and purple autotrophic bacteria were constructed for specific bioremediation applications. When the mats were challenged with metals, production and secretion of metal-binding extracellular polysaccharide bioflocculants were observed. The concentration of these negatively charged polysaccharides was correlated with the removal of manganese from the water column beneath a surface microbial mat. Bioflocculants from an Oscillatoria sp. that was isolated from the mat were collected and concentrated for characterization. A chromatographic analysis revealed a heterogeneous population of polysaccharides with respect to charge density and molecular size. The subpopulation of polysaccharides which exhibited the highest level of flocculating activity was polyanionic and had a molecular weight of more than 200,000. A glycosyl analysis of the bioflocculants revealed the presence of galacturonic acid (2.2%) and glucuronic acid (1.86%). The presence of these components, which were negatively charged at the pH levels generated by the mats during photosynthesis (pH > 7.5), may account for the metal-binding properties of the mats.     

Bioflocculants’ production in a biomass-degrading bacterium using untreated corn stover as carbon source and use of bioflocculants for microalgae harvest

Background

Bioflocculation has been developed as a cost-effective and environment-friendly method to harvest multiple microalgae. However, the high production cost of bioflocculants makes it difficult to scale up. In the current study, low-cost bioflocculants were produced from untreated corn stover by a biomass-degrading bacterium Pseudomonas sp. GO2.

Results

Pseudomonas sp. GO2 showed excellent production ability of bioflocculants through directly hydrolyzing various biomasses. The untreated corn stover was selected as carbon source for bioflocculants’ production due to its highest flocculating efficiency compared to that when using other biomasses as carbon source. The effects of fermentation parameters on bioflocculants’ production were optimized via response surface methodology. According to the optimal model, an ideal flocculating efficiency of 99.8% was obtained with the fermentation time of 130.46 h, initial pH of 7.46, and biomass content of 0.64%. The relative importance of carboxymethyl cellulase and xylanase accounted for 51.8% in the process of bioflocculants’ production by boosted regression tree analysis, further indicating that the bioflocculants were mainly from the hydrolysates of biomass. Biochemical analysis showed that it contained 59.0% polysaccharides with uronic acid (34.2%), 32.1% protein, and 6.1% nucleic acid in the bioflocculants, which had an average molecular weight as 1.33 × 10⁶ Da. In addition, the bioflocculants showed the highest flocculating efficiency at a concentration of 12.5 mg L^?1 and were stable over broad ranges of pH and temperature. The highest flocculating efficiencies obtained for Chlorella zofingiensis and Neochloris oleoabundans were 77.9 and 88.9%, respectively.

Conclusions

The results indicated that Pseudomonas sp. GO2 can directly utilize various untreated lignocellulolytic biomasses to produce low-cost bioflocculants, which showed the high efficiency to harvest two green microalgae in a low GO2 fermentation broth/algal culture ratio.

     

Evaluation of the flocculation potential and characterization of bioflocculant produced by Micrococcus sp. Leo

Bioflocculants are safe, biodegradable and environmentally friendly biopolymeric materials. These merits portend it as preferred alternative to inorganic and organic synthetic polymeric flocculants. The culture conditions optimal for the production of bioflocculant by Micrococcus sp. Leo with subsequent evaluation of the properties of the produced compound were investigated. Optimum culture conditions for bioflocculant production included 2% (vol/vol) inoculum size, incubation temperature of 28°C, agitation speed of 160 rpm and initial pH of 4.0. Glucose and (NH₄)₂SO₄ and Al³⁺ were the best as sole carbon, nitrogen and cation sources, respectively. The purified bioflocculant flocculated kaolin suspension optimally at a dosage of 0.2 mg/mL following jar test, and flocculating activity of about 70% was retained after heat treatment of 100°C. Chemical analysis showed that the bioflocculant was composed of 28.4% polysaccharide, 2.6% protein and 9.7%. uronic acid. Thermogravimetric analysis demonstrated that the bioflocculant could not decompose completely at 400°C. FTIR spectra revealed the presence of hydroxyl, carboxyl and amino groups as the main functional groups. The bioflocculant produced by Micrococcus sp. Leo appears to hold promise as an alternative to conventional flocculants commonly used in water/wastewater treatment.     

Novel bioflocculant produced by salt-tolerant,alkaliphilic strain Oceanobacillus polygoni HG6 and its application in tannery wastewater treatment

The optimized production of MBF-HG6, which is a novel salt-tolerant alkaliphilic bioflocculant produced by Oceanobacillus polygoni with its application in tannery wastewater treatment was investigated in this study. It was found the optimal carbon source, nitrogen source, cation, and initial pH of the medium for bioflocculant production were starch, urea, Fe²⁺, and pH 9.0, respectively. The best stability in the temperature range was from 0 to 80°C and the purified MBF-HG6 contained polysaccharides of 81.53% and proteins of 9.98%. The carboxyl, hydroxyl, and amino groups were determined in bioflocculants, while the optimized bioflocculating activity was observed as 90.25% for the dosages of 6.96mL MBF-HG6, 4.77mL CaCl₂ (1%, m/v), and 19.24g/L NaCl using response surface methodology. In addition, SS and turbidity removal rates of the tannery wastewater (4g/L MBF-HG6) could, respectively, reach 46.49% and 91.08%, indicating that the great potential was emerged in enhancement of tannery wastewater treatment by MBF-HG6.     

Characterization of a compound bioflocculant produced by mixed culture of <Emphasis Type="Italic">Rhizobium radiobacter</Emphasis> F2 and <Emphasis Type="Italic">Bacillus sphaeicus</Emphasis> F6

A compound bioflocculant CBF-F26, produced by mixed culture of Rhizobium radiobacter F2 and Bacillus sphaeicus F6, was investigated with regard to its physicochemical and flocculating properties. It was identified as a polysaccharide bioflocculant composed of rhamnose, mannose, glucose, and galactose, respectively, in a 1.3: 2.1: 10.0: 1.0 molar ratio. The average molecular weight was determined as 4.79 × 10⁵ Da by gel-permeation chromatography. Infrared spectrum and X-ray photoelectron spectroscopy revealed the presence of carboxyl, hydroxyl and amino groups in its structure. Thermostability test suggested that CBF-F26 was thermostable and high flocculating activity was maintained. Thermogravimetric property, intrinsic viscosity and surface morphology of CBF-F26 were also studied. CBF-F26 was effective under neutral and weak alkaline conditions (pH 7.0–9.0), and flocculating activities of higher than 90% were obtained in the concentration range of 8–24 mg l⁻¹ at pH 8.0. The flocculation could be stimulated by cations Ca²⁺, Zn²⁺, Fe²⁺, Al³⁺, and Fe³⁺. In addition, the probable flocculation mechanisms were proposed.     

Characterization of bioflocculant MBF3-3 produced by an isolated <Emphasis Type="Italic">Bacillus </Emphasis>sp.

Summary  The characteristics of bioflocculant MBF3-3 produced by Bacillus sp. BF3-3 were investigated here. MBF3-3 showed excellent flocculating activity on real and synthetic wastewaters, and consumed a much lower dosage than that of the widely used polyaluminum chloride (PAC) when flocculating brewery wastewater. Except Fe³⁺, metal ions, including Al³⁺, Mg²⁺, Ca²⁺, K⁺ and Na⁺, can stimulate the flocculating activity of MBF3-3 obviously, and the stimulating effects increased in the order: monovalent < bivalent < trivalent. MBF3-3 was mainly composed of acidic polysaccharide (66.1%) and protein (29.3%), in which acidic polysaccharide was the main effective flocculating component. OH and COO– groups may play a vital role in the flocculation of suspended particles.     

Flocculation properties of xanthan produced by Xanthomonas campestris

Summary Xanthan had a flocculating activity in a kaolin suspension and high flocculating activity was obtained in the suspension (pH 7.0) adding Al³⁺, Fe³⁺ or Fe²⁺. Xanthan had high flocculating activity not only in other inorganic suspensions such as active carbon and acid clay but also in organic suspensions of cellulose and yeast. From these flocculation properties, xanthan is anticipated to be utilized in wide areas as a new biodegradable, harmless biopolymer flocculant.     

Characterization and flocculating properties of an extracellular biopolymer produced from a Bacillus subtilis DYU1 isolate

Biopolymer DYU500 produced from Bacillus subtilis DYU1 was found to have excellent flocculating ability. With the addition of 40 mg-DYU500/L and 50 mM CaCO₃, the optimum temperature for flocculation performance of DYU500 was 30 °C, giving the highest flocculating activity and rate of 13.5 and 97%, respectively. Analysis with Fourier transform infrared spectrophotometry (FT-IR), nuclear magnetic resonance spectrometry (NMR) and amino acid identification shows that the DYU500 biopolymer mainly possesses the structure of poly-glutamic acid (PGA). The average molecular weight of DYU500 was about (3.16–3.20) × 10⁶ Da as determined by gel permeation chromatography. The major components of biopolymer DYU500 were total sugars, uronic acids, proteins and polyamides (homopolymer of glutamic acid), accounting for a weight ratio of approximate 14.9, 2.7, 4.4 and 48.7% (w/w), respectively. The flocculating activity of DYU500 in the kaolin suspension was markedly stimulated by the addition of bivalent cations Ca²⁺ or Mg²⁺ in optimum concentration ranges of about 0.15–0.90 and 0.10–0.90 mM, respectively. The synergistic effect of cations was most effective at a weak acidic or neutral pH (6.0–7.0). The flocculating activity of DYU500 linearly decreased with an increase in incubation temperature and the activity was completely lost when heating upon 120 °C, arising from the destruction of the polyamides structure of DYU500. Moreover, mechanisms describing the flocculation process with DYU500 were proposed based on the experimental observations.     

Production,Characterization, and Flocculation Mechanism of Cation Independent,pH Tolerant,and Thermally Stable Bioflocculant from Enterobacter sp. ETH-2

Synthetic high polymer flocculants, frequently utilized for flocculating efficiency and low cost, recently have been discovered as producing increased risk to human health and the environment. Development of a more efficient and environmentally sound alternative flocculant agent is investigated in this paper. Bioflocculants are produced by microorganisms and may exhibit a high rate of flocculation activity. The bioflocculant ETH-2, with high flocculating activity (2849 mg Kaolin particle/mg ETH-2), produced by strain Enterobacter sp. isolated from activated sludge, was systematically investigated with regard to its production, characterization, and flocculation mechanism. Analyses of microscopic observation, zeta potential and ETH-2 structure demonstrates the bridging mechanism, as opposed to charge neutralization, was responsible for flocculation of the ETH-2. ETH-2 retains high molecular weight (603 to 1820 kDa) and multi-functional groups (hydroxyl, amide and carboxyl) that contributed to flocculation. Polysaccharides mainly composed of mannose, glucose, and galactose, with a molar ratio of 1∶2.9∶9.8 were identified as the active constituents in bioflocculant. The structure of the long backbone with active sites of polysaccharides was determined as a primary basis for the high flocculation activity. Bioflocculant ETH-2 is cation independent, pH tolerant, and thermally stable, suggesting a potential fit for industrial application.     

Characteristics of a biopolymer flocculant produced by Bacillus sp. PY-90

A biopolymer flocculant-producing bacterium, strain PY-90, was isolated and considered to belong to Bacillus subtilis. For the production of biopolymer flocculant by strain PY-90, a medium containing 2 to 5% l-glutamic acid as a nitrogen source was suitable. The biopolymer flocculant was a homopolymer composed of glutamic acid residues and was presumed to be poly(γ-glutamic acid). In kaolin suspension, the highest flocculating activity was attained at the biopolymer flocculant concentration of 20 mg/l. The flocculating activity was increased by the addition of Ca²⁺, and the optimum concentration of which was about 2 to 8 mM. The flocculating activity was high in an acidic pH range of 3.0 to 5.0, and decreased upon heating at 100°C.     

Study on the flocculability of the Arthrobacter sp., an actinomycete resuscitated from the VBNC state

A bioflocculant with high flocculating activity, LC13-SF, produced by strain LC13^T which was in a viable but nonculturable (VBNC) state, and which was woken up by Rpf (resuscitation promoting factor), was systematically investigated with regard to its fermentation conditions and flocculating activity. The key parameters influencing the bioflocculant LC13-SF were investigated through measuring the optical density at 660 (OD₆₆₀) of the fermentation liquid and the optical density at 550 (OD₅₅₀) of the centrifugal supernatant. The flocculating efficiency and the Zeta potentials were chosen as the response variables for the study of the flocculating activity. The results showed that the optimal conditions for bioflocculant LC13-SF production were a fermentation time of 72 h, an initial pH of 7.0, a fermentation temperature of 30°C and a shaking speed of 150 r/min. The optimized flocculating process was as follows: a final volume percentage of bioflocculant LC13-SF and 0.5% (w/w) CaCl₂ were 1.5 and 5%, respectively in a 4 g/L Kaolin suspension, and the system pH was adjusted to 8.0. Under these conditions, the flocculating efficiency and the absolute value of the Zeta potential reached 94.83% and 4.37, respectively.     

Removal of Mercury from Chloralkali Electrolysis Wastewater by a Mercury-Resistant Pseudomonas putida Strain

A mercury-resistant bacterial strain which is able to reduce ionic mercury to metallic mercury was used to remediate in laboratory columns mercury-containing wastewater produced during electrolytic production of chlorine. Factory effluents from several chloralkali plants in Europe were analyzed, and these effluents contained total mercury concentrations between 1.6 and 7.6 mg/liter and high chloride concentrations (up to 25 g/liter) and had pH values which were either acidic (pH 2.4) or alkaline (pH 13.0). A mercury-resistant bacterial strain, Pseudomonas putida Spi3, was isolated from polluted river sediments. Biofilms of P. putida Spi3 were grown on porous carrier material in laboratory column bioreactors. The bioreactors were continuously fed with sterile synthetic model wastewater or nonsterile, neutralized, aerated chloralkali wastewater. We found that sodium chloride concentrations up to 24 g/liter did not inhibit microbial mercury retention and that mercury concentrations up to 7 mg/liter could be treated with the bacterial biofilm with no loss of activity. When wastewater samples from three different chloralkali plants in Europe were used, levels of mercury retention efficiency between 90 and 98% were obtained. Thus, microbial mercury removal is a potential biological treatment for chloralkali electrolysis wastewater.     

Lipase from Pseudomonas aeruginosa LP602: biochemical properties and application for wastewater treatment

Lipase from Pseudomonas aeruginosa LP602, a bacterial strain isolated from a domestic wastewater sample, was preliminarily characterized. The enzyme exhibited maximum lipolytic activity at pH 8.0 where it was also stably maintained. At 55°C, the lipase had the highest activity but not stability. The enzyme was insensitive to EDTA and to many ions tested except Zn²⁺. It was sensitive to SDS but not to Tween-20, Tween-80 or Triton X-100. The enzyme was active towards a number of commercial food grade fats and oils. A suitable medium formula for lipase production was MMP containing 6.25% whey as a carbon source, 1% soybean oil as inducer and 0.5% yeast extract supplement. The culture was fed with glucose to a final concentration of 0.1% at the 15th hour of incubation. Lipase production under this condition was 3.5 U ml⁻¹. Both P. aeruginosa LP602 cells and the lipase were shown to be usable for lipid-rich wastewater treatment. Received 21 April 1998/ Accepted in revised form 6 August 1998     

Bioflocculant production by Bacillus sp. Gilbert isolated from a marine environment in South Africa

In our previous study we reported on the bioflocculant production by a Bacillus species isolated from sediment samples of Algoa Bay in the Eastern Cape Province of South Africa. In the current study we carried out further evaluation on the effect of different culture conditions on the bioflocculant production, as well as characterised the bioflocculant produced in detail. The bacteria produced bioflocculant optimally under the following conditions: using sodium carbonate (95.2% flocculating activity) and potassium nitrate (76.6% flocculating activity) as carbon and nitrogen sources, respectively; inoculum size of 3% (v/v); initial pH 9.0; and Al³⁺ as coagulant aid. The crude bioflocculant retained 44.2% residual flocculating activity after heating at 100°C for 15 min. Chemical analysis of the Bacillus sp. Gilbert purified bioflocculant demonstrated that it was composed mainly of polysaccharide. Fourier transform infrared spectroscopy analysis revealed the presence of hydroxyl, carboxyl and methylene groups in the bioflocculant and energy-dispersive X-ray analysis detected the elemental composition in mass proportion (% w/w) of C, N, O, S and P as 4.12:7.40:39.92:3.00:13.91. Scanning electron micrograph image of the bioflocculant revealed an amorphous compound.     

Harvesting the microalgae Phaeodactylum tricornutum with polyaluminum chloride, aluminium sulphate, chitosan and alkalinity-induced flocculation

The purpose of this study was to explore efficient methods of harvesting the microalga Phaeodactylum tricornutum. Natural sedimentation experiments, performed at different light and temperature conditions, did not yield significant improvements in efficiency even after 1?week. When alkalinity-induced flocculation was performed, both the flocculation efficiency and the concentration factor dramatically improved at pH?=?9.75 (0.5–0.7 units over the original pH of the culture) after 10?min settling time. Sedimentation rates are documented at pH ranging between pH?9.75 and 11.0. The results of the application of two conventional flocculants used in wastewater treatment, polyaluminium chloride and aluminium sulphate, are also presented. Chitosan was also used as a natural flocculating agent to improve possible contamination problems in the downstream process. pH was adjusted in order to determine optimum flocculation efficiency of chitosan in combination with a high concentration factor. Satisfactory results were found with chitosan at an adjusted pH of 9.9 using concentrations as low as 20?mg?L^?1, after testing a flocculant range of 5–200?mg?L^?1.     

Distribution of chitin/chitosan-like bioflocculant-producing potential in the genus Citrobacter

Some strains belonging to the genera Citrobacter and Enterobacter have been reported to produce chitin/chitosan-like bioflocculants (BFs) from acetate. In this study, to investigate the distribution of the BF-producing potential in the genus Citrobacter and to screen stably and highly BF-producing strains, we obtained 36 Citrobacter strains from different culture collection centers, which were distributed among seven species in the genus, and tested for the flocculating activities of their culture supernatants using a kaolin suspension method. As a result, 21 strains belonging to C. freundii (17 strains in 23 strains tested), C. braakii (two in two), C. youngae (one in one), and C. werkmanii (one in two) showed flocculating activity, but this ability was limited to cells grown on acetate. Gas chromatography/mass spectrometry (GC/MS) analysis of the hydrolysates from the BFs of five selected strains indicated that they consisted of glucosamine and/or N-acetylglucosamine, such as the chitin/chitosan-like BF (BF04) produced by Citrobacter sp. TKF04 (Fujita et al. J Biosci Bioeng 89: 40–46, 2000). Gel filtration chromatography using a high-performance liquid chromatography system revealed that the molecular weight ranges of these BFs varied, but the average sizes were all above 1.66?×?10⁶?Da.