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Evidence for a lectin in Kluyveromyces sp. that is involved in co-flocculation with Schizosaccharomyces pombe.

Co-flocculation is the aggregation of yeasts belonging to different genera or species. Kluyveromyces bulgaricus and Kluyveromyces lactis 5c are self-flocculent, but they can also co-flocculate with the non-flocculent yeast Schizosaccharomyces pombe 972 h(-). This co-flocculation is inhibited by D-galactose and galactose derivatives and involves the binding of a galactose-specific proteinic receptor (or lectin) of Kluyveromyces sp. to the cell wall galactomannans of S. pombe. The proteinic receptor is strongly anchored in the cell wall, it was partially purified by affinity chromatography using immobilized S. pombe galactomannans. This galactose-specific proteinic receptor does not appear to interfere in K. bulgaricus or K. lactis self-flocculation, which is mediated by another galactose-specific lectin weakly linked at the cell wall.

Binding, Competitive↗

The identification, characterization, and mapping of a gene for flocculation in Saccharomyces sp.

Genetic studies of a flocculent haploid strain of Saccharomyces have revealed the flocculation to be dominant and controlled at a single gene locus. The flocculation character of both hybrids and haploids derived from such hybrids appears to be influenced by the repression or derepression status of the culture. Mapping studies of this flocculation gene have revealed that it is linked to ade 1 and therefore located on Chromosome I. Consequently, this is a different gene to the three flocculation genes studied by other laboratories because they have found such genes to be unlinked to ade 1. The flocculation gene being discussed in this paper has been designated FLO4. FLO4 has been located 32-33 cM from the Chromosome I centromere and 37 cM from ade 1 (i.e., FLO4 is on the opposite side of the centromere to ade 1).

Chromosome Mapping↗

Use of electron microscopy to characterize the surfaces of flocculent and nonflocculent yeast cells.

The surfaces of flocculent and nonflocculent yeast cells have been examined by electron microscopy. Nonextractive preparative procedures for scanning electron microscopy allow comparison in which sharp or softened images of surface details (scars, etc.) are the criteria for relative abundance of flocculum material. Asexually flocculent budding-yeast cells cannot be distinguished from nonflocculent budding-yeast cells in scanning electron micrographs because the scar details of both are well resolved, being hard and sharp. On the other hand, flocculent fission-yeast cells are readily distinguished from nonflocculent cells because fission scars are mostly soft or obscured on flocculent cells, but sharp on nonflocculent cells. Sexually and asexually flocculent fission-yeast cells cannot be distinguished from one another as both are heavily clad in "mucilaginous" or "hairy" coverings. Examination of lightly extracted and heavily extracted flocculent fission-yeast cells by transmission electron microscopy provides micrographs consistent with the scanning electron micrographs.

Flocculation↗

Physicochemical properties of cross-linked poly-gamma-glutamic acid and its flocculating activity against kaolin suspension.

Cross-linked poly-gamma-glutamic acid (C-L gamma-PGA) was prepared with gamma-PGA irradiated with gamma-ray at various kGy values. The physicochemical properties including viscosity and water absorption capacity were compared between C-L gamma-PGA and several typical flocculating agents. The viscosity of C-L gamma-PGA increased with the dose of gamma-irradiation, although the water absorption capacity of C-L gamma-PGA did not, which showed a maximum of 1005.6 ml/g at 20 kGy. Flocculating activity against kaolin suspension was not observed for any of the test compounds when used singly. However, the activity of C-L gamma-PGA markedly increased following the addition of polyaluminum chloride. The activity increased with temperature up to 80 degrees C and remained at 80 degrees C of heat pretreatment for 1 h, but did not at more than 50 degrees C of heat pretreatment for 24 h. The activity was also observed within a pH range of 4.5-10.0. Both the water absorption capacity and flocculating activity of C-L gamma-PGA decreased in parallel with increasing NaCl concentration, suggesting that the flocculating activity of C-L gamma-PGA was associated with its water absorption capacity, rather than viscosity. An investigation of the effects of various cations on the flocculating activity of C-L gamma-PGA showed that only trivalent cations had a synergistic effect. The mechanism of C-L gamma-PGA flocculating activity is discussed based on the results of preliminary experiments.

Adsorption↗

[Performance of a novel combined flocculant HECES].

Combined flocculants with higher environmental safety and lower ecological risk are urgently required in the process of water and wastewater treatment. HECES, a kind of flocculants with high effect and high ecological safety was synthesized by natural and biodegradable polysaccharide starch of corn and inorganic aluminium salt. Flocculent performance includes sediment character and turbidity removal rate. The jar test results showed that the flocculent performance of HECES with the dose of 3.0 mg.L-1 was prior to the combined adding 4.5 mg.L-1 PAC and 1.0 mg.L-1 PAM in treating kaolin suspensions. The optimal concentration of HECES was 8.0 mg.L-1 and 4.0 mg.L-1 in treating domestic wastewater and municipal effluent respectively, with the comparison to 50% an 40% of PAC, the removal rate of turbidity reached 95% and 99%, respectively. It was also identified that the optimal concentration required to effect flocculation depended on kaolin concentration, and on the character of the wastewater within the range examined. It could be more effective to treat wastewater with higher concentration of HECES. The flocculating performance would be better at pH 4.0-9.5. The superior ecological safety of HECES was due to its lower dosage and its lower concentration of residual activated aluminium.

Flocculation↗

Performance of basaltic dust issued from an asphaltic plant as a flocculant additive for wastewater treatment.

The feasibility of using basaltic dust as a flocculant additive or coagulant aid for wastewater treatment was assessed in this research. The experimental study was divided into two stages: 1) physicochemical characterisation of the basaltic dust by applying standardised techniques, and 2) evaluation of this material as flocculant additive for the coagulation-flocculation of wastewater treated for reuse. Coagulation-flocculation experiments were carried out in the laboratory with a mixture of industrial and municipal wastewater samples collected from two points of the final discharge of the Mexico City sewerage system. Aluminium sulphate and lime were used as coagulants and the basaltic dust as flocculant additive, by applying the jar-test technique. The results of the corrosivity, reactivity, explosiveness, toxicity, inflammability and biological risk tests indicated that this material is classified as a non-hazardous waste (according to the Mexican legislation, NOM-052-ECOL-1993). The density, oxide content and particle size values of basaltic dust were similar to those reported for the flocculant additive denominated activated silica. The jar test results showed a positive effect of basaltic dust over the effluent and sludge qualities, to the extent that coagulant doses can be reduced 30% (from 150 mg/L to 110 mg/L of Al2(SO4)3).

Construction Materials↗

Screening of flocculant-producing strains by NTG mutagenesis.

Screening of new microorganism being able to produce efficiently flocculants was carried out. A new model for screening efficient flocculant-producing strains was designed and tested. The results showed that this model for screening efficient flocculant-producing strains is very reliable and can greatly shorten the screening period. 13 flocculant-producing strains were isolated from activated sludge by conventional method. A strain, designated as HHE6, produced the bioflocculant with the turbidity removal 98% for kaolin suspension. Six of 13 strains selected as the original strains were treated with NTG as mutagen, and five mutant strains (HHE-P7, HHE-A8, HHE-P21, HHE-P24, HHE-A26) with high flocculation efficiency was obtained by selection, which exhibited the flocculation rate for kaolin suspension above 90%. Strains HHE6, HHE-P7, and HHE-P24 were classified as Penicillium purpurogenum, HHE-P21 as Penicillium cyclopium, HHE-A26 as Aspergillus versicolor and HHE-A8 as Aspergillus fumigatus, and it is hitherto unreported for biofloccutant-producing strains of Penicillium. The growth of the six strains (HHE6, HHE-P7, HHE-A8, HHE-P21, HHE-P24, HHE-A26) had similar curves, i.e. firstly increasing rapidly, keeping relatively constant then and finally decreasing gradually with cultivation time. The production of bioflocculants by strains showed the similar pattern to strain growth.

Bacteria↗

[Advances in research and application of water-treatment flocculants].

Water-treatment flocculants are being developed from traditional to inorganic, from inorganic to organic, from natural organic to synthetic organic, from synthetic organic to modified natural organic, from inorganic or organic to inorganic-organic, and from chemical to biological with ecological safety. In order to promote the rapid development and practical application of water-treatment flocculants, the research progress and application situation of flocculants for water treatment at home and abroad were reviewed. In particular, the characteristics of two important flocculant types, chemical and microbiological flocculants, as well as their application processes in water treatment were summarized, their development orientations were analyzed and evaluated, and the future research directions aimed to overcome the current shortages in developing and applying flocculants in China were put forward.

Ecosystem↗

Flocculation of sulfamerazine suspensions by a cationic polymer.

Flocculation by a cationic polymer of sulfamerazine suspensions containing a wetting agent was evaluated. Suspensions with sufficient surfactant concentrations to ensure complete wetting were deflocculated. When the anionic surfactant, dioctyl sodium sulfosuccinate, was used as a wetting agent, the suspensions were flocculated over a limited polymer concentration range. Flocculation was attributed to simultaneous interaction of a polymer molecule with more than one particle. At higher polymer concentrations, the particles were covered completely with polymer, leading to repulsion between the particles and deflocculation of the suspensions. The polymer concentration required for flocculation provided evidence for interaction between the anionic surfactant and the cationic polymer. Suspensions containing a nonionic surfactant also were flocculated using various polymer concentrations. When a surfactant mixture was employed in the suspensions, the peak sedimentation volume of flocculated systems and the concentration of polymer at the peak depended on the surfactant mixture composition.

Cations↗

Review: the dominant flocculation genes of Saccharomyces cerevisiae constitute a new subtelomeric gene family.

The quality of brewing strains is, in large part, determined by their flocculation properties. By classical genetics, several dominant, semidominant and recessive flocculation genes have been recognized. Recent results of experiments to localize the flocculation genes FLO5 and FLO8, combined with the in silicio analysis of the available sequence data of the yeast genome, have revealed that the flocculation genes belong to a family which comprises at least four genes and three pseudogenes. All members of this gene family are located near the end of chromosomes, just like the SUC, MEL and MAL genes, which are also important for good quality baking or brewing strains. Transcription of the flocculation genes is repressed by several regulatory genes. In addition, a number of genes have been found which cause cell aggregation upon disruption or overexpression in an as yet unknown manner. In total, 33 genes have been reported that are involved in flocculation or cell aggregation.

Amino Acid Sequence↗

Flocculation behavior of asphaltenes in solvent/nonsolvent systems.

Diffusion coefficients of asphaltenes dissolved in two aromatic solvents, toluene-d(8) or ethylbenzene-d(10), were measured with the pulsed-field gradient spin echo nuclear magnetic resonance (PFG-SE NMR) technique upon addition of flocculant (pentane-d(12) or heptane-d(16)). It was observed that the change in the diffusion coefficients, as a function of amount of added flocculant, was small in the concentration interval studied (up to 30 wt% alkane). Complementary kinetic flocculation studies were made at alkane additions above 55 wt%. The initial change in turbidity upon the addition of alkane was measured with an UV-VIS spectrophotometer. The obtained stability ratio, W, showed that asphaltenes were least stable in the ethylbenzene-pentane system and most stable in the toluene-heptane system. These findings were in agreement with the PFG-SE NMR. When combining the results from the two different techniques it appeared as if there was a dramatic increase in flocculation above a certain "threshold concentration" of added alkane. Furthermore, the flocculation appeared to be reaction controlled until as much as 63 wt% of n-pentane or, alternatively, 68 wt% of n-heptane had been added to the systems, after which the flocculation became primarily diffusion controlled. Finally, careful relaxation measurements showed that the asphaltenes displayed two distinctly different transverse (T(2)) relaxation times (most probably averages), one at 0.6 ms and the other at 7 ms.

Journal Article↗

Comparative extraction procedures for a galactose-specific lectin involved in flocculation of Kluyveromyces lactis strains.

The extraction of a lectinic factor involved in yeast flocculation, from two Kluyveromyces lactis strains (a flocculent K. lactis 5c and a non-flocculent K. lactis 5a strain) was performed using EDTA and two surfactants, Hecameg and HTAC. The properties of the different extracts were tested by haemagglutination and reflocculation of deflocculated K. lactis 5c cells. Hecameg gave the highest yields of active lectinic extract but the extraction with EDTA seemed more specific. HTAC extracts showed a very low activity. The possibilities of extraction of the agglutinating factor, either by an ion chelator or by surfactants, suggest that this factor may be anchored in the cell envelope, i.e. the cell wall and the membrane, by different mechanisms. All the assays revealed a galactose-specific lectinic activity was present that in the flocculent as well as in the non-flocculent strain. This indicates that the absence of flocculation with K. lactis 5a is mainly due to a defect in the ligands of the lectin rather than to a loss of the lectinic factor itself.

Calcium↗

Temperature influence of nonionic polyethylene oxide and anionic polyacrylamide on flocculation and dewatering behavior of kaolinite dispersions.

Nonionic polyethylene oxide (PEO) and anionic polyacrylamide (PAM) flocculation of kaolinite dispersions has been investigated at pH 7.5 in the temperature range 20-60 degrees C. The surface chemistry (zeta potential), particle interactions (shear yield stress), and dewatering behavior were also examined. An increase in the magnitude of zeta potential of kaolinite particles, in the absence of flocculant and at a fixed PEO and PAM concentration, with increasing temperature was observed. The zeta potential behavior of the flocculated particles indicated a decrease in the adsorbed polymer layer thickness, while at the same time, however, the adsorbed polymer density showed a significant increase with increasing temperature. These results suggest that polymer adsorption was accompanied by temperature-influenced conformation changes. The hydrodynamic diameter and supernatant solution viscosity of both polymers decreased with increasing temperature, consistent with a change in polymer-solvent interactions and conformation, prior to adsorption. The analysis of the free energy (DeltaG(ads)) of adsorption showed a strong temperature dependence and the adsorption process to be more entropically than enthalpically driven. The polymer conformation change and increased negative charge at the kaolinite particle surface with increasing temperature resulted in decreased polymer bridging and flocculation performance. Consequently, the shear yield stress and the rate and the extent of dewatering (consolidation) of the pulp decreased significantly at higher temperatures (>40 degrees C). The temperature effect was more pronounced in the presence of PEO than PAM, with 40 and 20 degrees C indicated as the optima for enhanced performance of the latter and former flocculants, respectively. The results demonstrate that a temperature-induced conformation change, together with polymer structure type, plays an important role in flocculation and dewatering behavior of kaolinite dispersions.

Journal Article↗

Organo-modified cationic silica nanoparticles/anionic polymer as flocculants.

The synthesis of quaternized silica nanoparticles and its application to fine clay flocculation were investigated. N-trimethoxysilylpropyl-N,N,N-trimethylammonium chloride was used as a cationic reagent to introduce quaternary amine groups onto the surfaces of silica nanoparticles via the formation of covalent bonds between the methoxy groups of the cationic reagents and the silanol groups in the silica surface. The zeta potential, zeta, and charge density of the silica particles modified under various reaction conditions were determined. Dynamic clay flocculation experiments using a photometric dispersion analyzer (PDA) showed that the cationic silica alone contributed little to the flocculation. However, the cationic silica, in conjunction with an anionic polymer of high M(w) and low charge density, led to a significant improvement in the flocculation of fine clay particles. The mechanism of flocculation was explored by a systematic investigation of interaction between cationic silica and anionic polymers as well as of their adsorption behavior on clay surfaces. The influence of factors such as pH and electrolyte concentration on clay flocculation was also investigated.

Journal Article↗

Preparation of high activity whole cell biocatalyst by permeabilization of recombinant flocculent yeast with alcohol.

Flocculent yeast Saccharomyces cerevisiae YF234 (MATa ura3-52 trp1Delta2 his ade 2-1 can1-100 sta1 FLO8) cells overexpressing glyoxalase I and having strong flocculation ability were permeabilized with isopropyl alcohol and ethanol under various conditions. The treatment with 40% isopropyl alcohol significantly improves the initial reaction rates of recombinant flocculent yeast cells. Moreover, the reactivity of permeabilized flocculent yeast cells was similar to that of dispersed cells with EDTA. On the other hand, the flocculation ability of yeast cells was not affected by the treatment with alcohol solutions of various concentrations and treatment time length. Therefore, the recombinant flocculent yeast cells permeabilized with alcohol are very effective whole cell biocatalysts.

Journal Article↗

Characterization of Macromolecular Flocculants Produced by Phormidium sp. Strain J-1 and by Anabaenopsis circularis PCC 6720.

Several benthic cyanobacteria were found to produce significant amounts of extracellular flocculants. The macromolecular flocculants produced by Phormidium sp. strain J-1 and Anabaenopsis circularis PCC 6720 were characterized. The Phormidium flocculant is a sulfated heteropolysaccharide to which fatty acids and protein are bound. The polysaccharide backbone is composed of uronic acids, rhamnose, mannose, and galactose. The A. circularis flocculant is also an acidic polysaccharide containing keto acid residues and neutral sugars, but to which no fatty acids, proteins, or sulfates are linked. Both flocculants could be recovered from growth medium by precipitation with cetyltrimethylammonium bromide and were found to bind the cationic dye Alcian-blue in a linear proportion to their concentration in solution. The latter property was used to quantify flocculant concentrations in culture supernatants and natural water samples and to compute their anion densities.

Journal Article↗

Repression and induction of flocculation interactions in Saccharomyces cerevisiae.

The biological control of flocculation interactions by factors related to growth under different conditions of aeration was documented with a new assay for flocculence. The degree of flocculence expressed in a genetically defined Saccharomyces cerevisiae strain (FLO1/FLO1 ade1/ade1) remained constant during aerobic growth but varied with aeration. Flocculence was repressed in anaerobically growing cells but was induced in stationary cells or cells returned to aerobic growth. Repression was correlated with the selective inactivation of cell surface lectin-like components. The changes in flocculence were accompanied by changes in 16 extractable proteins separated by electrophoresis; however, a clear correlation between specific protein bands and flocculence could not be established. The study clearly demonstrated that the phenotypic expression of FLO1 could be reproducibly manipulated for experimental purposes by aeration alone.

Aerobiosis↗

Fungal fimbriae. III. The effect on flocculation in Saccharomyces.

Flocculent strains of Saccharomyces cerevisiae and S. carlsbergensis (S. uvarum) produced many short (0.5 mum) hairs (fimbriae) on the outer cell walls. Non-flocculent strains produce few fimbriae. Cells that are flocculent in wort but not in defined medium produce fimbriae only in the former medium. Cells treated with pronase lose both their fimbriae and the ability to flocculate. Cells treated with alpha-amylase retain some fimbriae but lose the ability to flocculate. It is suggested that the fimbriae may be the surface mannan-protein complexes known to be involved in flocculation and also in protein secretion. Haploid cells of both mating types also produced fimbriae, some of which apparently have a terminal knob.

Amylases↗