Guanghui Yu

Insights into extracellular polymeric substances of cyanobacterium Microcystis aeruginosa using fractionation procedure and parallel factor analysis

Investigations on the extracellular polymeric substances (EPS) are crucial for better understanding the growth and proliferation of cyanobacterium Microcystis aeruginosa. In this study, a combined approach of fractionation procedure and parallel factor (PARAFAC) analysis were applied to characterize the EPS of M. aeruginosa. Physicochemical analysis showed that the contents of polysaccharides in EPS matrix were higher than those of proteins, regardless of the differences in growth phases and nutritional levels in medium. Organic matters were mainly distributed in the tightly bound EPS (TB-EPS) fraction during the exponential growth phase, whereas they sharply released to the soluble EPS (SL-EPS) and loosely bound EPS (LB-EPS) fractions at the decay period. Fluorescence excitation-emission matrix (EEM) was applied to characterize the specific compositions in EPS matrix, and all the fluorescence EEM spectra collected could be successfully decomposed into a four-component model by PARAFAC analysis. Component 1 [excitation/emission (Ex/Em) = 220/340], component 2 (Ex/Em = 280/340) and component 3 [Ex/Em = (200, 220, 270)/296] were attributed to protein-like substances, while component 4 [Ex/Em = (250, 340)/438] belonged to humic-like substances. Pearson correlation analysis demonstrated that tryptophan-like substances in the LB-EPS and TB-EPS fractions were positively correlated with Microcystis growth, whereas in the SL-EPS fraction, tryptophan-like as well as humic-like substances were associated with the growth of M. aeruginosa. The scientific implication for Microcystis growth and proliferation, based on the results of fractionation procedure and EEM-PARAFAC analysis, was also presented.

Binding of organic ligands with Al(III) in dissolved organic matter from soil: implications for soil organic carbon storage.

The binding characteristics of organic ligands with Al(III) in soil dissolved organic matter (DOM) is essential to understand soil organic carbon (SOC) storage. In this study, two-dimensional (2D) FTIR correlation spectroscopy was developed as a novel tool to explore the binding of organic ligands with Al(III) in DOM present in soils as part of a long-term (21-year) fertilization experiment. The results showed that while it is a popular method for characterizing the binding of organic ligands and metals, fluorescence excitation-emission matrix-parallel factor analysis can only characterize the binding characteristics of fluorescent substances (i.e., protein-, humic-, and fulvic-like substances) with Al(III). However, 2D FTIR correlation spectroscopy can characterize the binding characteristics of both fluorescent and nonfluorescent (i.e., polysaccharides, lipids, and lignin) substances with Al(III). Meanwhile, 2D FTIR correlation spectroscopy demonstrated that the sequencing/ordering of organics binding with Al(III) could be modified by the use of long-term fertilization strategies. Furthermore, 2D FTIR correlation spectroscopy revealed that the high SOC content in the chemical plus manure (NPKM) treatment in the long term fertilization experiment can be attributed to the formation of noncrystalline microparticles (i.e., allophane and imogolite). In summary, 2D FTIR correlation spectroscopy is a promising approach for the characterization of metal-organic complexes.

Functional Groups Determine Biochar Properties (pH and EC) as Studied by Two-Dimensional 13C NMR Correlation Spectroscopy

While the properties of biochar are closely related to its functional groups, it is unclear under what conditions biochar develops its properties. In this study, two-dimensional (2D) 13C nuclear magnetic resonance (NMR) correlation spectroscopy was for the first time applied to investigate the development of functional groups and establish their relationship with biochar properties. The results showed that the agricultural biomass carbonized to biochars was a dehydroxylation/dehydrogenation and aromatization process, mainly involving the cleavage of O-alkylated carbons and anomeric O-C-O carbons in addition to the production of fused-ring aromatic structures and aromatic C-O groups. With increasing charring temperature, the mass cleavage of O-alkylated groups and anomeric O-C-O carbons occurred prior to the production of fused-ring aromatic structures. The regression analysis between functional groups and biochar properties (pH and electrical conductivity) further demonstrated that the pH and electrical conductivity of rice straw derived biochars were mainly determined by fused-ring aromatic structures and anomeric O-C-O carbons, but the pH of rice bran derived biochars was determined by both fused-ring aromatic structures and aliphatic O-alkylated (HCOH) carbons. In summary, this work suggests a novel tool for characterising the development of functional groups in biochars.

Multiple fluorescence labeling and two dimensional FTIR-13C NMR heterospectral correlation spectroscopy to characterize extracellular polymeric substances in biofilms produced during composting.

Knowledge on the structure and function of extracellular polymeric substances (EPS) in biofilms is essential for understanding biodegradation processes. Herein, a novel method based on multiple fluorescence labeling and two-dimensional (2D) FTIR-(13)C NMR heterospectral correlation spectroscopy was developed to gain insight on the composition, architecture, and function of EPS in biofilms during composting. Compared to other environmental biofilms, biofilms in the thermophilic (>55 °C) and cooling (mature) stage of composting have distinct characteristics. The results of multiple fluorescence labeling demonstrated that biofilms were distributed in clusters during the thermophilic stage (day 14), and dead cells were detected. In the mature stage (day 26), the biofilm formed a continuous layer with a thickness of approximately 20-100 μm around the compost, and recolonization of cells at the surface of the compost was easily observed. Through 2D FTIR-(13)C NMR correlation heterospectral spectroscopy, the following trend in the ease of the degradation of organic compounds was observed: heteropolysaccharides > cellulose > amide I in proteins. And proteins and cellulose showed significantly more degradation than heteropolysaccharides. In summary, the combination of multiple fluorescence labeling and 2D correlation spectroscopy is a promising approach for the characterization of EPS in biofilms.

Combination of two-dimensional correlation spectroscopy and parallel factor analysis to characterize the binding of heavy metals with DOM in lake sediments

Enhanced knowledge on the binding of heavy metal (HM) with dissolved organic matter (DOM) is essential for understanding the toxicity and migration of HMs. In this study, two-dimensional correlation spectroscopy (2D-COS) and parallel factor (PARAFAC) analysis were combined to characterize the metal binding properties of DOMs, which were respectively extracted from macrophyte- and algal-dominant sediments (named MDOM and ADOM) in a eutrophic shallow lake. 2D absorption COS revealed that MDOM exhibited more HM binding sites (193, 195, 196, 199, 201, 203, 205, 207, 208, 212, 217 nm) than ADOM (201, 205 nm). PARAFAC analysis identified one protein- and two humic-like components from all titrated samples, with each component exhibiting different binding behaviors. The modified Stern-Volmer model showed that PARAFAC-derived components in MDOM had higher conditional stability constants (logKM) than in ADOM, suggesting that macrophyte-dominant sediments might play a more important role in the detoxification of HMs. Meanwhile, low binding abilities of Zn(II)-DOM complexes indicated that the toxicity of zinc in eutrophic lakes should not be overlooked. More aromatic functional groups and binding sites were suggested to be responsible for the high binding ability. 2D-COS was a better approach than PARAFAC analysis for exploring HM-DOM interaction.

Different analysis techniques for fluorescence excitation–emission matrix spectroscopy to assess compost maturity

Assessment of compost maturity is essential for achieving high quality compost. In this study, fluorescence excitation-emission matrix spectroscopy combined with different analysis techniques was applied to improve the sensitivity of compost maturity assessment. Results showed that composts in two parallel piles could be believed mature after 37d when combined with the evolution of temperature, chemical and biological indices in the two piles. Pearson correlation between the common maturity indices and fluorescence analysis parameters demonstrated that fluorescence regional integration (FRI) had a higher correlation coefficient than that of fluorescence intensities and the ratios of peaks, suggesting that FRI technique is more suitable to characterize the maturity of compost than the other two analysis techniques, i.e., peak intensity and peak ratio. Furthermore, the fluorescence spectroscopy combined with FRI analysis could be used as a valuable industrial and research tool for assessing compost maturity.

In situ observation of the growth of biofouling layer in osmotic membrane bioreactors by multiple fluorescence labeling and confocal laser scanning microscopy.

Since the concept of the osmotic membrane bioreactor (OMBR) was introduced in 2008, it has attracted growing interests for its potential applications in wastewater treatment and reclamation; however, the fouling mechanisms of forward osmosis (FO) membrane especially the development of biofouling layer in the OMBR are not yet clear. Here, the fouled FO membranes were obtained from the OMBRs on days 3, 8 and 25 in sequence, and then the structure and growing rule of the biofouling layer formed on the FO membrane samples were in-situ characterized by multiple fluorescence labeling and confocal laser scanning microscopy (CLSM). CLSM images indicated that the variations in abundance and distribution of polysaccharides, proteins and microorganisms in the biofouling layer during the operation of OMBRs were significantly different. Before the 8th day, their biovolume dramatically increased. Subsequently, the biovolumes of β-d-glucopyranose polysaccharides and proteins continued increasing and leveled off after 8 days, respectively, while the biovolumes of α-d-glucopyranose polysaccharides and microorganisms decreased. Extracellular polymeric substances (EPS) played a significant role in the formation and growth of biofouling layer, while the microorganisms were seldom detected on the upper fouling layer after 3 days. Based on the results obtained in this study, the growth of biofouling layer on the FO membrane surface in the OMBR could be divided into three stages. Initially, EPS was firstly deposited on the FO membrane surface, and then microorganisms associated with EPS located in the initial depositing layer to form clusters. After that, the dramatic increase of the clusters of EPS and microorganisms resulted in the quick growth of biofouling layer during the flux decline of the OMBR. However, when the water flux became stable in the OMBR, some microorganisms and EPS would be detached from the FO membrane surface.

Investigation on extracellular polymeric substances from mucilaginous cyanobacterial blooms in eutrophic freshwater lakes

Enhanced knowledge on extracellular polymeric substances (EPSs) of mucilaginous cyanobacterial blooms could improve our understanding of its ecological significance. This study for the first time investigated the extraction and fractionation of EPS matrix from cyanobacterial blooms in a eutrophic freshwater lake, and the changes in chemical compositions in EPS matrix during extraction were systematically investigated by two-dimensional correlation spectroscopy (2D-COS). The analyses demonstrated that organic matters were unevenly distributed among the EPS matrix, with most of organic matters being tightly bound to cyanobacterial cells. In addition, the soluble and loosely bound EPS fractions mainly consisted of proteins, while polysaccharides became the predominant compounds in the tightly bound EPS fraction. Heating extraction at 60°C for 30min led to a high EPS yield and low cell lysis when compared with other extraction methods. The 2D-COS results revealed a preferential release of OH in polysaccharides versus amide I in proteins in the initial heating; whereas further extension of heating resulted in EPS degradation, with degradation rates arranging in a decreased order from amide I, amide II, polysaccharides-like substances to polysaccharides. These results obtained would help enhance our insights into EPS characterization from cyanobacterial blooms in eutrophic lakes.

Production and characterization of acidophilic xylanolytic enzymes from Penicillium oxalicum GZ-2.

Multiple acidophilic xylanolytic enzymes were produced by Penicillium oxalicum GZ-2 during growth on wheat straw, rice straw, corn stover, and wheat bran. The expression of xylanase isoforms was dependent on substrate type and nitrogen source. The zymograms produced by the SDS-PAGE resolution of the crude enzymes indicated that wheat straw was the best inducer, resulting in the highest xylanase (115.2U/mL) and β-xylosidase (89mU/mL) activities during submerged fermentation. The optimum temperature and pH for xylanase activity were 50°C and 4.0, respectively; however, the crude xylanase enzymes exhibited remarkable stability over a broad pH range and showed more than 90% activity at 50°C for 30min at pH 4.0-8.0. The results revealed that P. oxalicum GZ-2 is a promising acidophilic xylanase-producing microorganism that has great potential to be used in biofuels, animal feed, and food industry applications.

Insights into complexation of dissolved organic matter and Al(III) and nanominerals formation in soils under contrasting fertilizations using two-dimensional correlation spectroscopy and high resolution-transmission electron microscopy techniques

Understanding the organomineral associations in soils is of great importance. Using two-dimensional correlation spectroscopy (2DCOS) and high resolution-transmission electron microscopy (HRTEM) techniques, this study compared the binding characteristics of organic ligands to Al(III) in dissolved organic matter (DOM) from soils under short-term (3-years) and long-term (22-years) fertilizations. Three fertilization treatments were examined: (i) no fertilization (Control), (ii) chemical nitrogen, phosphorus and potassium (NPK), and (iii) NPK plus swine manure (NPKM). Soil spectra detected by the 2DCOS Fourier transform infrared (FTIR) spectroscopy showed that fertilization modified the binding characteristics of organic ligands to Al(III) in soil DOM at both short- and long- term location sites. The CH deformations in aliphatic groups played an important role in binding to Al(III) but with minor differences among the Control, NPK and NPKM at the short-term site. While at the long-term site both C-O stretching of polysaccharides or polysaccharide-like substances and aliphatic O-H were bound to Al(III) under the Control, whereas only aliphatic O-H, and only polysaccharides and silicates, were bound to Al(III) under NPK and NPKM, respectively. Images from HRTEM demonstrated that crystalline nanominerals, composed of Fe and O, were predominant in soil DOM under NPK, while amorphous nanominerals, predominant in Al, Si, and O, were dominant in soil DOM under Control and NPKM. In conclusion, fertilization strategies, especially under long-term, could affect the binding of organic ligands to Al(III) in soil DOM, which resulted in alterations in the turnover, reactivity, and bioavailability of soil organic matter. Our results demonstrated that the FTIR-2DCOS combined with HRTEM techniques could enhance our understanding in the binding characteristics of DOM to Al(III) and the resulted nanominerals in soils.