Surface properties of bacteria from activated sludge in relation to bioflocculation

Two bacterial stains were isolated from the activated sludge and identified as Leucobacter sp. and Alcaligenesfaecalis by 16S rDNA sequencing. Pure cultures of these two strains, representing well or poorly settled bacteria, were used to investigate the mechanism of bioflocculation in activated sludge. Based on the analyses of the characteristics of cells hydrophobicity, ζ-potential, flocculation ability and extracellular polymeric substance （EPS） composition under different growth stages, it was found that the ratio of cell EPS protein had the highly influence on ζ-potential and hydrophobicity, which were important factors to bioflocculation. Cellulase and Proteinase K could destroy the extracellular biopolymer and resulted in a decrease in the hydrophobicity and ζ-potential. However, in our study, the flocculation characteristics exhibited differently in relation to cellulase and Proteinase K. Flocculation of cells treated with cellulase and Proteinase K decreased sharply, and then recovered quickly in cellulase treatment, while cells treated with Proteinase K showed no sign of recovery. This reveals that the presence of protein in extracellular biopolymer plays an important role to the bioflocculation of cells.

Production and application of a novel bioflocculant by multiple-microorganism consortia using brewery wastewater as carbon source

The flocculating activity of a novel bioflocculant MMF1 produced by multiple-microorganism consortia MM1 was investigated. MM1 was composed of strain BAFRT4 identified as Staphylococcus sp. and strain CYGS1 identified as Pseudomonas sp. The flocculating activity of MMF1 isolated from the screening medium was 82.9%, which is remarkably higher than that of the bioflocculant produced by either of the strains under the same condition. Brewery wastewater was also used as the carbon source for MM1, and the cost-effective production medium for MM1 mainly comprised 1.0 L brewery water （chemical oxygen demand （COD） 5000 mg/L）, 0.5 g/L urea, 0.5 g/L yeast extract, and 0.2 g/L （NH4）2SO4. The optimal conditions for the production of MMF1 was inoculum size 2%, initial pH 6.0, cultivating temperature 30℃, and shaking speed 160 r/min, under which the flocculating activity of the MMF1 reached 96.8%. Fifteen grams of purified bioflocculant could be recovered from 1.0 L of fermentation broth. MMF1 was identified as a macromolecular substance containing both protein and polysaccharide. It showed good flocculating performance in treating indigotin printing and dyeing wastewater, and the maximal removal efficiencies of COD and chroma were 79.2% and 86.5%, respectively.

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.

Characteristics and Culture Conditions of a Bioflocculant Produced by Penicillium sp.

Objective To study the characteristics of a bioflocculant named MBF7 produced by Penicillum strain HHE-P7 and the effects of cultivation conditions on bioflocculant production. Methods The chemical group in the bioflocculant molecules was shown by Fourier transform infrared （FTIR） spectra, and the average molecular weight of MBF7 was estimated by gel permeation chromatography. The effects of medium components on bioflocculant production and flocculating activity were studied. Results Phospho-, amino-, hydroxyl, and carboxyl groups were the major fractions of MBF7, and the molecule weight was about 3.0 × 10^5 Da. In addition, the carbon and nitrogen sources favorable for the bioflocculant production were glucose and yeast extract respectively. When the initial pH of the medium was adjusted to 5.0, high flocculant efficiency could be achieved. Conclusion The bioflocculant MBF7 is a new macromolecule with high flocculating efficiency for Kaolin suspension, and could be produced under appropriate culture conditions.

Two-stage pH Control Mode in Batch Fermentation of a Novel Bioflocculant from Corynebacterium Glutamicum

The effect of pH of the fermentation medium on cell growth and the production of a novel bioflocculant(named REA-11) by Corynebacterium glutamicum CCTCC M201005 were investigated. The maximum biomass(2.23 g/L) and flocculating activity(142.2 U/mL) were simultaneously obtained at the 14th hour when the pH value of the culture medium was maintained at 7.0 during the whole fermentation process. The production of REA-11 kept on a trend of increase till the later phase of fermentation process, which resulted in the ultimate flocculating activity of the culture broth to enhance to nearly 100 U/mL at pH 6.0. A two-stage pH control mode was adopted in REA-11 production in which the pH value of the culture medium was controlled at 7.0 during the first 14 h, then decreased to 6.0 that was maintained until the end of the fermentation process. With the two-stage pH control mode, the maximum flocculating activity reached 178.8 U/mL which was 30% higher than that obtained under the condition of pH 7.0 and the biomass enhanced about 15%. Compared with the fermentation process without pH control, REA-11 production and cell growth via the two-stage pH control mode increased 80% and 25%, respectively.

Screening and flocculating properties of bioflocculant-producing microorganisms

Screening of bioflocculant-producing microorganisms was carded out. A strain that secreted excellent bioflocculant was isolated from municipal sewage using the spread plate technique, identified as Klebsiella sp. by the analytical profile index （API） identification system, and named A9. Several important factors that had an effect on A9＇s bioflocculant-producing and flocculating activity were studied. A total of 4 g/L Kaolin suspension was used to measure the flocculating activity of the bioflocculant from A9. It was found that maltose and urea were Ag＇s best carbon and nitrogen sources, respectively, and the flocculating activity of the flocculating agent from A9 was markedly increased by the addition of trivalent cations such as Fe^3＋ and Al^3＋; furthermore, the bioflocculant produced by A9 was most effective when the pH value was 6.0.

Screening of Bioflocculant-Producing Strain by Ion Implantation and Flocculating Characteristics of Bioflocculants

A bioflocculant-producing mutator strain, NIM-192, was screened out through nitrogen ion implanting into F J-7 strain. The results showed that NIM-192 had good genetic stability and high flocculating activity, and the flocculating rate increased by 34.26% than that of the original. Sucrose, complex nitrogen source contained yeast extract, urea and pH 7.0～ 9.0 were chosen as the best carbon source, nitrogen source and initial solution pH for bioflocculant production, respectively. The bioflocculant kept high and stable flocculating activity at alkalinous reaction mixture with a pH beyond 7.0, while the flocculating activity was remarkably reduced when the reaction pH was lower than 7.0. Addition of many cations could obviously increase the flocculating rate, among which Ca^2＋ demonstrated the best effect. The bioflocculant had very strong acid-base stability and thermo-stability.The flocculating rate kept over 86% when pH of the bioflocculant was in a range of 3.0 ～ 12.0, and the change of flocculating activity was not great when heated at 100℃ for 60 rain.

Flocculating Properties and Production of the Compound Bioflocculant by Rhizobium Radiobacter F2 and Bacillus Sphaeicus F6

A compound bioflocculant CBF,produced by mixed culture of Rhizobium radiobacter F2 and Bacillus sphaeicus F6, was investigated with regard to its production and flocculating properties. The optimization of the culture medium constituents including carbon source,nitrogen source and C / N ratio,metal ions and ionic strength on CBF production were studied. Flocculating properties of CBF were examined by a series of experiments and CBF had good flocculating activities in kaolin suspension with divalent cations and stable over wide range of p H. Studies of the flocculating properties revealed that the flocculation could be stimulated by cations Ca2+,Mg2+,Fe2+,Al3+and Fe3+. In addition,it was stable at 4-30 ℃ in the presence of Ca Cl2. It was found to be effective for flocculation of a kaolin suspension under neutral and weak alkaline conditions( p H 7. 0- 9. 0),and flocculating activities of higher than 95% were obtained when the CBF concentrations among 6- 14 mg / L at p H 8. 0. The results of this study indicate that CBF is a potential replacement of conventional synthetic flocculants and is widely applied in water treatment and downstream processing of food and fermentation industries.

Screening and Characterization of a Bioflocculant Produced by Aeromonas sp

To isolate the bioflocculant-producing bacteria from activated sludge and investigate the flocculating characteristics of the newly isolated bioflocculant. Methods Bacteria were screened from activated sludge samples to isolate bioflocculant-producing bacteria. Flocculating activity was used as a measure of the flocculating capability of the bioflocculant. Results A novel bioflocculant-producing bacterium was isolated, which was identified to belong to genus Aeromonas and named as Aeromonas sp. Nil. Flocculating activity increased in the presence of K, Na^＋, or Ca^2＋. The highest flocculating activities for kaolin suspension were obtained in acidic pH ranges, and optimum pHs for it were 3.0, 4.0, and 5.0 with 1 mmol/L K^＋, Ca^＋, and Na^＋ present, respectively. The highest flocculating activities for soil suspension were observed at pH 8.0. The bioflocculant had a good flocculating activity and could achieve a flocculating activity of 92.4% for kaolin suspension at a dosage of only 1 mg.L-1, and its activity in kaolin suspension was decreased by only 9.2% after heating at 100℃ for 60 min. Conclusion The bioflocculant produced by Aeromonas sp. Nil has strong flocculating activity and high stability, which affords high possibility of its practical use.

Studies on Bioflocculant Production by Pseudoalteromonas sp. NUM8, a Marine Bacteria Isolated from the Circulating Seawater

A bioflocculant producing potential bacteria was isolated from the circulating seawater of bio-filter using streak plate methods.The bacteria was identified through biochemical characteristics,partial 16S ribosomal ribonucleic acids(rRNA),nucleo-tide sequencing,and BLAST analysis of the gene sequence that showed the bacteria have 99%similarity to Pseudoalteromonas sp.and deposited in GenBank as Pseudoalteromonas sp.NUM8 with accession number JX435820.Influences of time course assay,carbon sources,nitrogen sources,inoculum size,as well as initial pH on the bacteria producing extracellular bioflocculant activity were investigated.The results showed that the strain optimal production period of microbial bioflocculant was at 72 h(flocculating activity of 94.5%),then dropped slowly.The bacteria optimally produced the bioflocculant when 1.0%sucrose and 0.5%sodium nitrate were used as sole sources of carbon and nitrogen with flocculating activities of 92.8%and 93.8%respectively.Also,the bacteria produced the bioflocculant optimally when initial pH of the medium was 5.0(flocculating activity 93.2%),and when Ca^(2+)was used as cation(flocculating activity 93.4%).The culture condition of inoculum size of 3%(v/v)was optimal flocculant pro-duction(flocculating activity 94.4%).Composition analyses indicated the bioflocculant to be principally a glycoprotein made up of about 34.3%protein and 63.4%total carbohydrate.

On an ecology-friendly treatment reagent for oily wastewater

Bioflocculants have received increasing attention in research because they are low cost,non-toxic,biodegradable,operationally simple and useful in many water and wastewater treatment applications.In this study,we focused mainly on the screening of the flocculant-producing strain and its application in oily wastewater treatment.Bacteria strain B-6-1 was isolated by the pyridine screening method.UPC-1 reagent produced by B-6-1 had efficient flocculating activity.It was deduced that the main component of UPC-1 reagent was polysaccharide through purification and analysis.The influence of dosage of reagent,pH value,temperature and oil concentration on the flocculation effect of UPC-1 reagent was investigated.

Screening of Seawater Compound Bioflocculant Producing Bacterial Strains and Optimization of Their Culture Medium

Compared with single bioflocculant,compound bioflocculant(CBF)normally has more stable and higher flocculation efficiencies.In order to produce a new compound bioflocculant,four bacterial strains with relative high rate of flocculation were selected from the coastal zone sediment,all of which were determined as Pseudomonas sp.by 16S rRNA sequencing.Through compounding,the ideal proportion of compound bioflocculant consisting of two bacterial strains(F5∶M5)was determined as 1∶2,resulting in 6%increase in terms of flocculation rate.The significant fermentation factors in culture media and their limitations were investigated via single factor experiments and Plackett⁃Burman design,obtaining four key variables,MgSO_(4),(NH_(4))_(2)SO_(4),sucrose and peptone,which were further optimized by response surface methodology(RSM)for their optimal concentrations.Finally,the ideal culture medium was determined,containing(NH_(4))_(2) SO_(4)0.523 g,MgSO_(4)3.7 g,sucrose 12.82 g,K_(2)HPO_(4)4 g,KH_(2)PO_(4)3 g,NaCl 33 g,and deionized water 1 L.The flocculation rate of experimental data(88.92)agreed well with the model prediction,which was 16.60 higher than original rate.

Effect of bioflocculants on the coagulation activity of alum for removal of trihalomethane precursors from low turbid water

Reactivity of chlorine towards hydrophobic groups present in natural organic matter（NOM）provokes the formation of carcinogenic disinfection byproducts such as trihalomethanes in chlorinated water. The present study aimed to investigate the variations in coagulant activity of alum using two different bioflocculants（coagulant aid） namely, Moringa oleifera and Cyamopsis tetragonoloba for the removal of hydrophobic fractions of NOM and subsequent chlorine consumption by treated water. Effect of dual coagulants on trihalomethane surrogate parameters such as total organic carbon, dissolved organic carbon, UV absorbing materials and prominent hydrophobic species such as phenolic groups along with aromatic chromophores, polyhydroxy aromatic moiety have also been studied. The concept of differential spectroscopy and absorbance slope index has been employed to understand the combined effects of alum-bioflocculants on the reactivity of NOM with chlorine. Our result shows that the combination of alum and C. tetragonoloba is more efficient for reducing trihalomethane surrogates from chlorinated water as compared to M. oleifera. C. tetragonoloba elicited synchronized effects of sweep coagulation and particle bridging-adsorption which eventually facilitated efficient removal of hydrophobic fractions of NOM. The variation in the mechanistic approach of bioflocculants was due to the presence of cationic charge on M. oleifera and adhesive property of C. tetragonoloba.

Optimized production of a novel bioflocculant M-C11 by Klebsiella sp. and its application in sludge dewatering

The optimized production of a novel bioflocculant M-C11 produced by Klebsiella sp. and its application in sludge dewatering were investigated. The optimal medium carbon source,nitrogen source, metal ion, initial pH and culture temperature for the bioflocculant production were glucose, NaNO3, MgSO4, and pH 7.0 and 25°C, respectively. A compositional analysis indicated that the purified M-C11 consisted of 91.2% sugar, 4.6% protein and 3.9% nucleic acids（m/m）. A Fourier transform infrared spectrum confirmed the presence of carboxyl, hydroxyl,methoxyl and amino groups. The microbial flocculant exhibited excellent pH and thermal stability in a kaolin suspension over a pH range of 4.0 to 8.0 and a temperature range of 20 to 60°C.The optimum bioflocculating activity was observed as 92.37% for 2.56 mL M-C11 and 0.37 g/L CaCl2 dosages using response surface methodology. The sludge resistance in filtration（SRF）decreased from 11.6 × 1012 to 4.7 × 1012m/kg, which indicated that the sludge dewaterability was remarkably enhanced by the bioflocculant conditioning. The sludge dewatering performance conditioned by M-C11 was more efficient than that of inorganic flocculating reagents,such as aluminum sulfate and polymeric aluminum chloride. The bioflocculant has advantages over traditional sludge conditioners due to its lower cost, benign biodegradability and negligible secondary pollution. In addition, the bioflocculant was favorably adapted to the specific sludge pH and salinity.