生物絮凝剂絮凝活性评价方法的探讨

对生物絮凝剂絮凝活性的评价方法进行了探讨。研究发现,在固定絮凝剂添加量的情况下仅仅比较絮凝率大小,从而对絮凝活性进行评价的方法是不科学的。絮凝剂絮凝活性应以絮凝率和絮凝剂最适添加量两个指标为其活性评价参数,在有较高絮凝率的情况下,絮凝剂添加量越少则絮凝活性越高。     

Production of an exopolysaccharide bioflocculant by Sorangium cellulosum

AIMS: To isolate a new exopolysaccharide bioflocculant produced by the myxobacterium Sorangium cellulosum NUST06, and to characterize its chemical composition and expolysaccharide production relative to carbon source. METHODS AND RESULTS: Exopolysaccharide levels and biomass production by S. cellulosum NUST06 were analysed relative to carbon source. Glucose in the medium at a level of 3 g l(-1) completely inhibited cell growth and exopolysaccharide production, but low concentrations of glucose (1-2 g l(-1)) could stimulate cell utilization of starch. The chemical composition and flocculating activity of the NUST06 exopolysaccharide was investigated. The flocculant comprised 38.3% proteins and 58.5% carbohydrates, of which glucose, mannose and glucuronic acid were present at 51.3%, 39.2% and 10.5%, respectively. The flocculating activity of the NUST06 flocculant depended strongly on cations. CONCLUSIONS: It is feasible to produce an exopolysaccharide bioflocculant by the strain NUST06 in a mineral salts medium using starch as a carbon source. SIGNIFICANCE AND IMPACT OF THE STUDY: This strain may be advantageous for commercial bioflocculant production and may enrich existing knowledge of myxobacteria.     

金属离子对生物絮凝剂REA-11合成的影响

考察了Mg2 、Na 、K 、Ca2 对谷氨酸棒杆菌生长及生物絮凝剂合成的影响。结果表明,培养基中添加适当的Mg2 对菌体的生长和絮凝剂的产生有一定的促进作用。K 和Ca2 对絮凝剂的产生具有一定的抑制作用。Na 的添加对菌体的生长和絮凝剂的产生均无显著影响。     

Microbial flocculant from Arcuadendron sp. TS-49

Summary A flocculant was purified from the culture broth of Archuadendron sp. TS-49 by a series of precipitations with acetone, 60% ammonium sulfate-butanol, salting-out by dialysis, and cetylpyridinium chloride. The flocculating activity was observed most highly at pH 3.0 and markedly enhanced by the addition of salts, especially in the case of FeCl₃ or FeSO₄. This bioflocculant efficiently flocculated all tested solids, including various microorganisms and organic/ inorganic materials. Qualitative analyses of the bioflocculant showed that it might contain hexosamine, uronic acid, neutral sugar, and protein.     

Simultaneous production of hydrogen and bioflocculant by Enterobacter sp. BY-29

Hydrogen and a bioflocculant could be produced simultaneously by anaerobic culture of Enterobacter sp. BY-29. For production of hydrogen and the bioflocculant by cell culture of the bacterium in batch cultures, cultivation at 37 °C in a medium containing glucose as a carbon source and Polypepton as a nitrogen source was found to be suitable. In continuous production of hydrogen and the bioflocculant by cell culture or immobilized cells of the bacterium, the hydrogen production rate and hydrogen yield by the immobilized cells on porous glass beads in stirred and column reactors were higher than those by the cell culture in a stirred reactor. However, production of the bioflocculant by the cell culture was superior to that by the immobilized cells in continuous production.     

Production of a novel bioflocculant MNXY1 by Klebsiella pneumoniae strain NY1 and application in precipitation of cyanobacteria and municipal wastewater treatment

Aims: To isolate and characterize the novel bioflocculant‐producing bacteria, to optimize the bioflocculant production and to evaluate its potential applications. Methods and Results: Klebsiella pneumoniae strain NY1, a bacterium that produces a novel bioflocculant (MNXY1), was selected on the chemically defined media. It was classified according to the 16S rRNA gene sequence, morphological and microscopic characteristics. MNXY1 was characterized to contain 26% protein and 66% total sugar. The constituent sugar monomers of MNXY1, revealed by NMR analysis, are glucose, galactose and quinovose. Favourable culture conditions for MNXY1 production were determined. Strain NY1 produces a high level (14.9 g l^?1) of MNXY1. MNXY1 is thermostable and tolerant to the extreme pH. It precipitated 54% of cyanobacteria from laboratory culture and 72% of the total suspended solids from raw wastewater. Conclusions: Strain NY1 was identified to produce a novel bioflocculant MNXY1. The outstanding performance of MNXY1 in practical applications and its availability in copious amounts make it attractive for further investigation and development for industrial scale applications. Significance and Impact of the Study: This is first report for the identification of a quinovose‐containing bioflocculant and application of a protein–polysaccharide complex bioflocculant in precipitation of cyanobacteria. These findings suggest that MNXY1 holds great potential for use in management of harmful algae and city wastewater treatment.     

Characterization and flocculation properties of biopolymeric flocculant (glycosaminoglycan) produced by Cellulomonas sp. Okoh

Aims

Bioflocculant production potential of an actinobacteria isolated from a freshwater environment was evaluated and the bioflocculant characterized.

Methods and Results

16S rDNA nucleotide sequence and BLAST analysis was used to identify the actinobacteria and fermentation conditions, and nutritional requirements were evaluated for optimal bioflocculant production. Chemical analyses, FTIR, 1H NMR spectrometry and SEM imaging of the purified bioflocculant were carried out. The 16S rDNA nucleotide sequences showed 93% similarities to three Cellulomonas species (strain 794, Cellulomonas flavigena DSM 20109 and Cellulomonas flavigena NCIMB 8073), and the sequences was deposited in GenBank as Cellulomonas sp. Okoh (accession number HQ537132 ). Bioflocculant was optimally produced at an initial pH 7, incubation temperature 30°C, agitation speed of 160 rpm and an inoculum size of 2% (vol/vol) of cell density 1·5 × 10⁸ cfu ml^?1. Glucose (88·09% flocculating activity; yield: 4·04 ± 0·33 g l^?1), (NH₄)₂NO₃ (82·74% flocculating activity; yield: 4·47 ± 0·55 g l^?1) and MgCl₂ (90·40% flocculating activity; yield: 4·41 g l^?1) were the preferred nutritional source. Bioflocculant chemical analyses showed carbohydrate, protein and uronic acids in the proportion of 28·9, 19·3 and 18·7% in CPB and 31·4, 18·7 and 32·1% in PPB, respectively. FTIR and 1H NMR indicated the presence of carboxyl, hydroxyl and amino groups amongst others typical of glycosaminoglycan. SEM imaging revealed horizontal pleats of membranous sheets closely packed.

Conclusion

Cellulomonas sp. produces bioflocculant predominantly composed of glycosaminoglycan polysaccharides with high flocculation activity.

Significance and Impact of the Study

High flocculation activity suggests suitability for industrial applications; hence, it may serve to replace the hazardous flocculant used in water treatment.     

The production of bioflocculants by Bacillus licheniformis using molasses and its application in the sugarcane industry

A bioflocculant produced by B. licheniformis was investigated with regard to a low-cost culture medium and its industrial application. Molasses replaced sucrose as the sole carbon source in bioflocculant fermentation. The optimum low-cost culture medium was determined to be composed of 20 g/L molasses, 0.4 g/L urea, 0.4 g/L NaCl, 0.2 g/L KH₂PO₄, 1.6 g/L K₂HPO₄, and 0.2 g/L MgSO₄. The bioflocculant from B. licheniformis was then applied to treat sugarcane-neutralizing juice to remove colloids, suspended particles, and coloring matters in a sugar refinery factory. The optimal operation conditions were a bioflocculant dosage of 21 U/mL, pH 7.3 and a heating temperature of 100°C. The color and turbidity of the sugarcane juice reached IU 1267 and IU 206, respectively, after clarification with the bioflocculant; these values were almost the same as those acquired following treatment with polyacrylamide (PAM), the most widely applied flocculant in sugar industries. These results suggest the great potential for use of bioflocculants in the sugar refinery process.     

一种富集高温栅藻Desmodesmus sp. F51的新型生物絮凝剂

[背景]海科贝特氏菌(Cobetia marina)可产生大量具有絮凝活性的胞外产物,可视为一种新型的生物絮凝剂。高温栅藻(Desmodesmus sp.F51)是一种具有较高叶黄素含量的微藻,被认为是一种新兴的叶黄素来源,但利用该生物絮凝剂高效富集高温栅藻的相关研究迄今尚未见报道。[目的]以高温栅藻为对象,研究该新型生物絮凝剂的絮凝效率,并对絮凝机理进行初步探讨。[方法]探索在不同生长阶段微藻培养液添加生物絮凝剂、添加量、絮凝时间、pH对絮凝效率的影响,分析生物絮凝剂的功能基团,并测定在不同pH条件下添加生物絮凝剂前后高温栅藻的Zeta电位变化,以及在显微镜下分析藻细胞在添加生物絮凝剂前后的形态。[结果]在高温栅藻生长至稳定期(pH 8.0)添加2 mL生物絮凝剂,絮凝15 min絮凝效果最佳,达82.1%。傅里叶红外光谱(fourier transform infrared spectroscopy,FTIR)显示了多糖及酰胺结构的特征吸收峰,由此推测生物絮凝剂主要是多糖的混合物,含有少量蛋白质。根据Bradford法测定絮凝剂中蛋白含量约为0.4%(质量比),通过苯酚-硫酸法测定总糖质量分数约为34.5%(质量比),与FTIR分析结果基本相符。生物絮凝剂在pH 4.0-11.0保持60%以上的絮凝效率,说明无论是酸性或是碱性条件下絮凝效率都较高,结合Zeta电位的分析表明,推测生物絮凝剂对高温栅藻的絮凝机理中占主导地位的可能是吸附架桥作用。[结论]该研究对微藻生物絮凝具有重要的理论和实践意义。     

Phormidium J-1 bioflocculant: production and activity

Phormidium J-1, a benthic filamentous cyanobacterium, isolated from a drainage channel, was found to produce a high molecular weight polymer. This substance can flocculate bentonite particles from suspensions. At the stationary phase of growth the cells excreted this bioflocculant into the surrounding medium. Production was enhanced by reduction of the calcium content in the growth medium or by increasing its EDTA content. Above the isoelectric point (pH 3.5) the bioflocculant is negatively charged. The presence of minimal concentrations of divalent cations in the reaction mixture is required for its flocculating activity. The production of bioflocculant could be of great importance in clarification of turbid water bodies, thus allowing light penetration to the sediment/water interface. Bioflocculant production and excretion was not found in several other benthic cyanobacteria.Dedicated to Prof. Dr. H.-G. Schlegel on the occasion of his 60th birthday     

Poly‐γ‐glutamic acid produced from Bacillus licheniformis CGMCC 2876 as a potential substitute for polyacrylamide in the sugarcane industry

As an environmentally friendly and industrially useful biopolymer, poly‐γ‐glutamic acid (γ‐PGA) from Bacillus licheniformis CGMCC 2876 was characterized by the high‐resolution mass spectrometry and ¹H NMR. A flocculating activity of 11,474.47 U mL^?1 obtained with γ‐PGA, and the effects of carbon sources, ions, and chemical properties (D‐/L‐composition and molecular weight) on the production and flocculating activity of γ‐PGA were discussed. Being a bioflocculant in the sugar refinery process, the color and turbidity of the sugarcane juice was IU 1,877.36 and IU 341.41 with 0.8 ppm of γ‐PGA, respectively, which was as good as the most widely used chemically synthesized flocculant in the sugarcane industry—polyacrylamide with 1 ppm. The γ‐PGA produced from B. licheniformis CGMCC 2876 could be a promising alternate of chemically synthesized flocculants in the sugarcane industry. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1287–1294, 2015     

Screening of flocculant-producing microorganisms and some characteristics of flocculants

Summary 27 strains of microorganisms including moulds, bacteria, actinomyces and yeasts, which produced substances that flocculated a kaolin clay, were screened. One strain, identified as Arthrobacter sp., secreted bioflocculant during growth. The flocculant (MF-6) was purified to electrophoretic homogeneity by precipitation with ethanol and gel chromatography. Its molecular weight was higher than 10⁶ Da. It could efficiently flocculate all suspended solids tested in aqueous solution and colored water like dye solution, and its flocculating ability was significantly enhanced by adding Ca²⁺, Mn²⁺, Mg²⁺, etc.     

利用污泥提取液制备微生物絮凝剂

以污水厂剩余污泥作为培养基原料,经过一系列处理,探索微生物絮凝剂产生菌的最适发酵培养基配方,结果表明,污泥预处理条件以pH 12碱解条件最优,碳氮源产出量最大,补加8 g/L葡萄糖后灭菌,微生物絮凝剂产生菌LLin6可正常产絮,絮凝率达91.55%。该结果为降低微生物絮凝剂的制备成本,并实现污泥的减量化和污泥资源化利用提供了基础。     

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.     

Flocculant Production by Paecilomyces sp. Taxonomic Studies and Culture Conditions for Production

Screening of microorganisms producing a new kind of flocculating substance was carried out. A fungus that produced a large amount of a strong microbial cell flocculant was isolated from soil. The fungus was classified as a Paecilomyces species. The substance produced showed very interesting and unique flocculation characteristics. It could efficiently flocculate all tested suspended solids in aqueous solution, and flocculation was practically not affected by ionic strength, pH or temperature.

The influence of components of the culture medium upon the flocculant production by Paecilomyces sp. 1-1 was studied. Addition of hydrolysates of casein such as casamino acid and polypeptone stimulated the production of the flocculant. A high concentration of calcium ions also significantly increased the mycelial growth and the production of the flocculant. Under the optimum cultural conditions, a 500-fold dilution of the culture filtrate could efficiently flocculate and sediment E. coli cells.     

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.     

Technical communication: modelling the rates of biodegradation,nitrogen fixation and uptake of fixed nitrogen during bioremediation of crude oil-contaminated environments

A biopolymer flocculant, produced by a haloalkalophilic Bacillus sp. I-471 was isolated from a sea water sample from the tidal mud flats surrounding the city of Inchon, Korea. The bioflocculant (EPS471) was produced in late logarithmic growth phase, recovered by cold ethanol precipitation of the cell-free supernatant and purified by cetylpyridinium chloride treatment. Chemical analyses of EPS471 indicated that it was an acidic polysaccharide containing neutral sugars, namely, galactose, fructose, glucose (approximate ratio of 5:2:1) and uronic acids as major and minor components, respectively. SEM studies revealed that the polymer had a porous structure with small pore size distribution indicating the compactness of the polymer. Spectroscopic analyses of the polymer by nuclear magnetic resonance (NMR) and Fourier-transform infrared (FT-IR) spectroscopy revealed the presence of carboxyl and hydroxyl groups typical for a polysaccharide and that the polymer was a heteroglycan. The degradation temperature (T _d) analyzed by thermogravimetry was 307 °C. The rheological analysis showed the pseudoplastic nature of the polymer with shear-thinning behaviour. The polymer EPS471 may find possible application as an polymer for environmental bioremediation and other biotechnological processes.     

Production of a novel bioflocculant by fed-batch culture of Citrobacter sp.

Production of a novel bioflocculant by a fed-batch culture of Citrobacter sp. TKF04 was investigated using acetic acid as a sole carbon source. Synthesis of the bioflocculant was favored by dissolved O₂ tension at 20% of air saturation and C/N ratio (mol acetic acid/mol ammonium) of 10:1 in the feed solution. Under optimal conditions, 4.6 g crude bioflocculant per liter broth was produced, whose flocculating activity was 22 300 units. This activity was 9 times higher than that of the control (only acetic acid was supplied).     

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.     

Production of a biopolymer flocculant from Bacillus licheniformis and its flocculation properties

Bacillus licheniformis CCRC 12826 produced extracellularly an excellent biopolymer flocculant in a large amount when it was grown aerobically in a culture medium containing citric acid, glutamic acid and glycerol as carbon sources. The biopolymer flocculant was an extremely viscous material with a molecular weight over 2 x 10(6) by gel permeation chromatography. It could be easily purified from the culture medium by ethanol precipitation. It was shown to be a homopolymer of glutamic acid by amino acid analysis and thin layer chromatography and presumed to be poly-glutamic acid (PGA). This bioflocculant efficiently flocculated various organic and inorganic suspensions. It flocculated a suspended kaolin suspension without cations, although its flocculating activity was synergistically stimulated by the addition of bivalent or trivalent cations Ca2+, Fe3+ and Al3+. However, the synergistic effects of metal cations were most effective at neutral pH ranges. The comparison of the flocculating activity between the present biopolymer and a commercial lower molecular weight product showed that the biopolymer of the present study had much higher activity. The high productivity and versatile applications of PGA make its development as a new biodegradable, harmless, biopolymer flocculant economical and advantageous.