Characterization of a microbial polysaccharide-based bioflocculant and its anti-inflammatory and pro-coagulant activity.

We describe a novel bioflocculant, MBF-15, which is an exopolysaccharide extracted from the alkaliphilic bacterium Paenibacillus jamilae. The biophysical characteristics of MBF-15 were determined using Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. MBF-15 was also evaluated for its biocompatibility by examining its inflammatory, coagulant, and hemostatic properties in vitro and in vivo. Pretreatment of peripheral blood mononuclear cells with MBF-15 inhibited lipopolysaccharide-stimulated expression of inducible nitric oxide synthase, production of nitric oxide, and secretion of pro-inflammatory cytokines, including tumor necrosis factor-α and interleukin-6. In addition, MBF-15 increased both mRNA and protein levels of the anti-inflammatory cytokines transforming growth factor-ß and IL-10. The hemocompatibility of MBF-15 was investigated by measuring the hemolysis ratio and clotting times. MBF-15 had high pro-thrombogenic activity but was not hemolytic. In a rat model, MBF-15 showed superior hemostatic properties compared with chitosan. Thus, MBF-15 offers a promising combination of anti-inflammatory and pro-coagulant properties that may be useful for hemostasis in a variety of clinical settings.

Evaluation of hemocompatibility and hemostasis of a bioflocculant.

Bioflocculants are widely used in a large variety of applications, because of their wide range of flocculation. There is limited information in the literature about the ability of bioflocculants to perform beneficial functions without having adverse effects on blood components. In this study, general hemocompatibility of a bioflocculant (MBF-06) was investigated in an in vitro system, measuring flocculation of plasma proteins, red blood cell integrity, blood cell counts, platelet activation, clot formation, complement activation, cytotoxicity and inflammatory response. Effects of MBF-06 included low plasma protein flocculation, low platelet activation, low inflammatory response, no hemolysis or cytotoxicity and increased expression of coagulation markers, including prothrombin fragments 1+2. These findings suggested that the rapid hemostasis caused by MBF-06 resulted, essentially, from humoral coagulation. Overall, our observations revealed the great potential of MBF-06 as a novel hemostatic agent.

Removal of cadmium by bioflocculant produced by Stenotrophomonas maltophilia using phenol-containing wastewater.

Bioflocculants have been applied in numerous applications including heavy metals removal. A major bottleneck for commercial application of bioflocculant is its high production cost. Phenol-containing wastewater are abundantly available. However, the toxic phenol inhibited the microbial activities in the subsequent fermentation processes. Consequently, strains that can secrete phenol-degrading enzymes and simultaneously produce bioflocculants through directly degrading the phenol are of academic and practical interests. A phenol-degrading strain, Stenotrophomonas maltophilia ZZC-06, which can produce the bioflocculant MBF-06 using phenol-containing wastewater, was isolated in this study. The effects of culture conditions including initial pH, dissolved oxygen, phenol concentration, inoculum size, and temperature on MBF-06 production were analyzed. The experimental results showed that over 90% flocculating activity was achieved when the phenol was used as a carbon source and 4.99 g/L of MBF-06 was achieved under the optimized condition: 2.0% dissolved oxygen, 800 mg/L phenol concentration, 10% inoculum size, an initial pH of 6.0, and a temperature of 30 °C. The bioflocculant MBF-06 contained 71.2% polysaccharides and 27.9% proteins. The feasibility of cadmium removal using MBF-06 was evaluated. The highest flocculating efficiency for cadmium was 81.43%. This study shows for the first time that Stenotrophomonas maltophilia ZZC-06 can directly convert phenol into a bioflocculant, which can be used to effectively remove cadmium.

Immobilization of pectinase onto Fe3O4@SiO2-NH2 and its activity and stability.

Fe3O4 magnetic nanometer particles (MNPS) with a diameter of 30-40nm and coated by ammoniated silicon dioxides were developed by sol-gel method. The X-ray diffraction (XRD) pattern indicates that the Fe3O4 is wrapped in the amorphous SiO2, and the Fourier-transform infrared spectroscopy (FT-IR) analysis confirmed the NH2 group had been successfully introduced onto the surface of Fe3O4@SiO2 nanoparticles. The main factors related to the immobilization process of pectinase onto the magnetic support were optimized by multi-factor orthogonal experiments, and the recommended combination is that 3% glutaraldehyde (coupling agent), pH=8.0, 50°C, and cross-linking time 8h. Scanning electron microscope (SEM) and FT-IR analyses confirmed that the pectinase had been immobilized onto the surface of the amino modified nanoparticles. The storage stability and reusability of the pectinase were enhanced by the immobilization that 64.4% of the total starting activity was retrieved after 7 recycling batches and only about 21.1% of the total activity was lost after 30-day storage. For the application of the immobilized pectinase, the optimal pH and temperature were 8.5 and 55°C, respectively.

Production of bioflocculants prepared from formaldehyde wastewater for the potential removal of arsenic.

A novel bioflocculant (MBF-79) prepared using formaldehyde wastewater as carbon resource was investigated in the study. The optimal conditions for bioflocculant production were determined to be an inoculum size of 7.0%, initial pH of 6.0, and formaldehyde concentration of 350 mg/L. An MBF-79 of 8.97 g/L was achieved as the maximum yield. Three main elements, namely C, H, and O, were present in MBF-79 with relative weigh percentages of 39.17%, 6.74%, and 34.55%, respectively. The Gel permeation chromatography analysis indicated that the approximate molecular weight (MW) of MBF-79 was 230 kDa. MBF-79 primarily comprised polysaccharide (71.2%) and protein (27.9%). Additionally, conditions for the removal of arsenic by MBF-79 were found to be MBF-79 at 120 mg/L, an initial pH 7.0, and a contact time 60 min. Under the optimal conditions, the removal efficiencies of arsenate (0.5 mg/L) and arsenite (0.5 mg/L) were 98.9% and 84.6%, respectively. Overall, these findings indicate bioflocculation offers an effective alternative method of decreasing arsenic during water treatment.

Bioflocculant production by Haloplanus vescus and its application in acid brilliant scarlet yellow/red removal.

A novel bioflocculant MBF057 produced by a salt-tolerant Haloplanus vescus HW0579 was investigated in this study. The effects of culture conditions such as initial pH, inoculum size, and chemical oxygen demand (COD) of K-acid wastewater on MBF0579 production were studied. The result showed that 8.09 g/L purified MBF0579 was extracted with the following optimized conditions: 780 mg/L COD of K-acid wastewater as carbon source, inoculum size 12.5%, and initial pH 7.0. The biopolymer contained 78.6% polysaccharides and 21.1% proteins. The highest flocculating rate of 81.86 and 95.07% for the COD and chroma of acid brilliant scarlet gelb rot (yellow/red, GR) dye wastewater were achieved at a dosage of 150 mg/L, pH 2.0 and contact time 100 min. Overall, these findings indicate bioflocculation offers an effective alternative method of decreasing acid brilliant scarlet GR during dye wastewater treatment.

Instability of chromosome number and DNA methylation variation induced by hybridization and amphidiploid formation between Raphanus sativus L. and Brassica alboglabra Bailey.

BACKGROUND: Distant hybridization can result genome duplication and allopolyploid formation which may play a significant role in the origin and evolution of many plant species. It is unclear how the two or more divergent genomes coordinate in one nucleus with a single parental cytoplasm within allopolyploids. We used cytological and molecular methods to investigate the genetic and epigenetic instabilities associated with the process of distant hybridization and allopolyploid formation, measuring changes in chromosome number and DNA methylation across multiple generations. RESULTS: F1 plants from intergeneric hybridization between Raphanus sativus L. (2n = 18, RR) and Brassica alboglabra Bailey (2n = 18, CC) were obtained by hand crosses and subsequent embryo rescue. Random amplification of polymorphic DNA (RAPD) markers were used to identify the F1 hybrid plants. The RAPD data indicated that the hybrids produced specific bands similar to those of parents and new bands that were not present in either parent. Chromosome number variation of somatic cells from allotetraploids in the F4 to F10 generations showed that intensive genetic changes occurred in the early generations of distant hybridization, leading to the formation of mixopolyploids with different chromosome numbers. DNA methylation variation was revealed using MSAP (methylation-sensitive amplification polymorphism), which showed that cytosine methylation patterns changed markedly in the process of hybridization and amphidiploid formation. Differences in cytosine methylation levels demonstrated an epigenetic instability of the allopolyploid of Raphanobrassica between the genetically stable and unstable generations. CONCLUSIONS: Our results showed that chromosome instability occurred in the early generations of allopolyploidy and then the plants were reverted to largely euploidy in later generations. During this process, DNA methylation changed markedly. These results suggest that, epigenetic mechanisms play an important role in intergeneric distant hybridization, probably by maintaining a genetic balance through the modification of existing genetic materials.

[Genomic in situ hybridization in intergeneric hybrids between Raphanus sativus and Brassica oleracea].

Genomic in situ hybridization (GISH) was applied to study the meiosis of F1 plants from intergeneric hybrids between radish (Raphanus sativus, 2n=18, RR) and cabbage (Brassica oleracea, 2n=18, CC). The result showed that its somatic cells had the expected chromosomes, RC, 2n=18; but the pollen mother cells (PMCs) were different. There were three main kinds of PMCs. The first one was RC (2n=18), and the mean chromosome pairing pattern was 14.87I+1.20II+0.04III+0.06IV on Diakinesis. GISH indicated that most bivalents resulted from chromosome pairing between radish and cabbage, and the nine chromosomes of R-genome were separated mostly in the ratio 5/4 and 6/3 at Anaphase, so the chromosome number and components in gametes were not in equilibrium and the gametes were sterile. The second was RRCC (2n=36) with normal chromosome pairing and separation, producing unreduced gametes. And the third was nullisomic of RRCC in PMCs (2n<36) GISH showed that some radish chromosomes were lost in those PMCs, and its gametes had nine cabbage chromosomes and partial radish chromosomes. The mechanism of this chromosome reduplication was discussed in this paper.