Chongjian Tang

Characterization and genomic analysis of kraft lignin biodegradation by the beta-proteobacterium Cupriavidus basilensis B-8

BackgroundLignin materials are abundant and among the most important potential sources for biofuel production. Development of an efficient lignin degradation process has considerable potential for the production of a variety of chemicals, including bioethanol. However, lignin degradation using current methods is inefficient. Given their immense environmental adaptability and biochemical versatility, bacterial could be used as a valuable tool for the rapid degradation of lignin. Kraft lignin (KL) is a polymer by-product of the pulp and paper industry resulting from alkaline sulfide treatment of lignocellulose, and it has been widely used for lignin-related studies.ResultsBeta-proteobacterium Cupriavidus basilensis B-8 isolated from erosive bamboo slips displayed substantial KL degradation capability. With initial concentrations of 0.5–6 g L-1, at least 31.3% KL could be degraded in 7 days. The maximum degradation rate was 44.4% at the initial concentration of 2 g L-1. The optimum pH and temperature for KL degradation were 7.0 and 30°C, respectively. Manganese peroxidase (MnP) and laccase (Lac) demonstrated their greatest level of activity, 1685.3 U L-1 and 815.6 U L-1, at the third and fourth days, respectively. Many small molecule intermediates were formed during the process of KL degradation, as determined using GC-MS analysis. In order to perform metabolic reconstruction of lignin degradation in this bacterium, a draft genome sequence for C. basilensis B-8 was generated. Genomic analysis focused on the catabolic potential of this bacterium against several lignin-derived compounds. These analyses together with sequence comparisons predicted the existence of three major metabolic pathways: β-ketoadipate, phenol degradation, and gentisate pathways.ConclusionThese results confirmed the capability of C. basilensis B-8 to promote KL degradation. Whole genomic sequencing and systematic analysis of the C. basilensis B-8 genome identified degradation steps and intermediates from this bacterial-mediated KL degradation method. Our findings provide a theoretical basis for research into the mechanisms of lignin degradation as well as a practical basis for biofuel production using lignin materials.

Performance of high-loaded ANAMMOX UASB reactors containing granular sludge

The performance of high-loaded anaerobic ammonium oxidizing (ANAMMOX) upflow anaerobic sludge bed (UASB) reactors was investigated. Two ANAMMOX reactors (R1 with and R2 without effluent recycling, respectively) were fed with relatively low nitrite concentration of 240 mg-N L(-1) with subsequent progressive increase in the nitrogen loading rate (NLR) by shortening the hydraulic retention time (HRT) till the end of the experiment. A super high-rate performance with nitrogen removal rate (NRR) of 74.3-76.7 kg-N m(-3) day(-1) was accomplished in the lab-scale ANAMMOX UASB reactors, which was 3 times of the highest reported value. The biomass concentrations in the reactors were as high as 42.0-57.7 g-VSS L(-1) with the specific ANAMMOX activity (SAA) approaching to 5.6 kg-N kg-VSS(-1) day(-1). The high SAA and high biomass concentration were regarded as the key factors for the super high-rate performance. ANAMMOX granules were observed in the reactors with settling velocities of 73-88 m h(-1). The ANAMMOX granules were found to contain a plenty of extracellular polymers (ECPs) such as 71.8-112.1 mg g-VSS(-1) of polysaccharides (PS) and 164.4-298.2 mg g-VSS(-1) of proteins (PN). High content of hemachrome (6.8-10.3 μmol g-VSS(-1)) was detected in the ANAMMOX granules, which is supposed to be attributed to their unique carmine color.

Synthesis of Core–Shell Magnetic Fe3O4@poly(m-Phenylenediamine) Particles for Chromium Reduction and Adsorption

Magnetic Fe3O4@poly(m-phenylenediamine) particles (Fe3O4@PmPDs) with well-defined core-shell structure were first designed for high performance Cr(VI) removal by taking advantages of the easy separation property of magnetic nanoparticles (MNPs) and the satisfactory adsorption property of polymers. Through controlling the polymerization on MNPs, directly coating was realized without the complicated premodification procedures. The particle property and adsorption mechanism were analyzed in details. Fe3O4@PmPDs exhibited tunable PmPD shell thickness from 10 to 100 nm, high magnetic (∼150 to ∼73 emu g(-1)) and facile separation property by magnet. The coating of PmPD significantly enhanced Cr(VI) adsorption capacity from 46.79 (bare MNPs) to 246.09 mg g(-1) (71.55% PmPD loading proportion), much higher than many reported composite adsorbents. The high Cr(VI) removal performance was attributed to the adsorption of Cr(VI) on protonated imino groups and the efficient reduction of Cr(VI) to Cr(III) by amine, followed by Cr(III) chelated on imino groups, which are spontaneous and endothermic. The Fe3O4@PmPDs have great potential in treating Cr(VI)-contaminated water.

Identification and quantification of anammox bacteria in eight nitrogen removal reactors.

Various studies have revealed anaerobic ammonium oxidation (anammox) as a very attractive alternative process suitable for nitrogen removal from wastewater. Here we investigated anammox bacteria in eight different nitrogen removal reactors. The diversity and abundance of anammox bacteria were determined by the 16S rRNA gene analysis, fluorescence in situ hybridization with specific probes and real-time quantitative PCR (qPCR). In these reactors, at least eight unique near full length anammox 16S rRNA gene sequences were detected, which were distributed over two genera; Candidati Brocadia and Kuenenia. FISH results confirmed that only one anammox bacterium dominated the community in each of the eight reactors investigated in this study. qPCR analysis revealed that anammox bacteria were present in seven of the reactors in the order of 10(9) cells/ml and 10(7) cells/ml in reactor A1. The dominant and divergent Brocadia-like anammox phylotype in one reactor represented a novel species for which we propose the name Candidatus Brocadia sinica. Taken together, these results indicated that a single seeding source could be used to seed anammox reactors designed to treat different types of wastewater, which could lead to a faster start-up of bioreactors.

Suppression of anaerobic ammonium oxidizers under high organic content in high-rate Anammox UASB reactor

The effect of organic matter on the nitrogen removal performance of anaerobic ammonium oxidation (Anammox) process was investigated in an upflow anaerobic sludge blanket (UASB) reactor fed with nitrogen loading rate of 13.92 kg Nm(-3)day(-1) at an HRT of 0.83 h. Mass balance showed that the heterotrophic denitrification prevailed in the UASB reactor, and became the dominant reactions when high influent COD/NO(2)(-)-N ratios of 2.92 were applied. The Anammox bacterial growth was significantly suppressed by denitrifying communities under high organic matter content due to the weaker competition for nitrite (electron acceptor) and living space. Long-term operation of the Anammox UASB reactor under relatively high organic content resulted in weak recovery performance.

Enhanced nitrogen removal from pharmaceutical wastewater using SBA-ANAMMOX process.

Efficient biological nitrogen removal from pharmaceutical wastewater has been focused recently. The present study dealt with the treatment of colistin sulfate and kitasamycin manufacturing wastewater through anaerobic ammonium oxidation (ANAMMOX). The biotoxicity assay on luminescent bacterium Photobacterium phosphoreum (T3 mutation) showed that the pharmaceutical wastewater imparted severe toxicity with a relative luminosity of 3.46% ± 0.45%. During long-term operation, the cumulative toxicity from toxic pollutants in wastewater resulted in the performance collapse of conventional ANAMMOX process. A novel ANAMMOX process with sequential biocatalyst (ANAMMOX granules) addition (SBA-ANAMMOX process) was developed by combining high-rate ANAMMOX reactor with sequential biocatalyst addition (SBA). At biocatalyst addition rate of 0.025 g VSS (L wastewater)(-1) day(-1), the nitrogen removal rate of the process reached up to 9.4 kg N m(-3) day(-1) in pharmaceutical wastewater treatment. The effluent ammonium concentration was lower than 50 mg N L(-1), which met the Discharge Standard of Water Pollutants for Pharmaceutical Industry in China (GB 21903-2008). The application of SBA-ANAMMOX process in refractory ammonium-rich wastewater is promising.

Influence of substrates on nitrogen removal performance and microbiology of anaerobic ammonium oxidation by operating two UASB reactors fed with different substrate levels

Both ammonium and nitrite act as substrates as well as potential inhibitors of anoxic ammonium-oxidizing (Anammox) bacteria. To satisfy demand of substrates for Anammox bacteria and to prevent substrate inhibition simultaneously; two strategies, namely high or low substrate concentration, were carefully compared in the operation of two Anammox upflow anaerobic sludge blanket (UASB) reactors fed with different substrate concentrations. The reactor working at relatively low influent substrate concentration (NO(2)(-)-N, 240 mg-NL(-1)) was shown to avoid the inhibition caused by nitrite and free ammonia. Using the strategy of low substrate concentration, a record super high volumetric nitrogen removal rate of 45.24 kg-Nm(-3) day(-1) was noted after the operation of 230 days. To our knowledge, such a high value has not been reported previously. The evidence from transmission electron microscopy (TEM) showed that the morphology and ultrastructure of the Anammox cells in both the reactor enrichments was different.

The structure, density and settlability of anammox granular sludge in high-rate reactors

Microscopic observation and settling test were carried out to investigate the structure, density and settlability of anammox granules taken from a high-rate upflow anaerobic sludge blanket (UASB) reactor. The results showed that the anammox granules were irregular in shape and uneven on surface, and their structure included granule, subunit, microbial cell cluster and single cell. The gas pockets were often observed in the anammox granules, and they originated from the obstruction of gas tunnel by extracellular polymer substances (EPSs) and the inflation of produced dinitrogen gas. The volume of gas pockets became larger with the increasing diameter of anammox granules, which led to the decreasing density and the floatation of anammox granules. The diameter of anammox granules should be controlled at less than 2.20mm to avoid the granule floatation. A hypothesized mechanism for the granulation and floatation of anammox biomass was proposed.

Anaerobic ammonium oxidation for treatment of ammonium-rich wastewaters

The concept of anaerobic ammonium oxidation (ANAMMOX) is presently of great interest. The functional bacteria belonging to the Planctomycete phylum and their metabolism are investigated by microbiologists. Meanwhile, the ANAMMOX is equally valuable in treatment of ammonium-rich wastewaters. Related processes including partial nitritation-ANAMMOX and completely autotrophic nitrogen removal over nitrite (CANON) have been developed, and lab-scale experiments proved that both processes were quite feasible in engineering with appropriate control. Successful full-scale practice in the Netherlands will accelerate application of the process in future. This review introduces the microbiology and more focuses on application of the ANAMMOX process.

The Increasing Interest of ANAMMOX Research in China: Bacteria, Process Development, and Application

Nitrogen pollution created severe environmental problems and increasingly has become an important issue in China. Since the first discovery of ANAMMOX in the early 1990s, this related technology has become a promising as well as sustainable bioprocess for treating strong nitrogenous wastewater. Many Chinese research groups have concentrated their efforts on the ANAMMOX research including bacteria, process development, and application during the past 20 years. A series of new and outstanding outcomes including the discovery of new ANAMMOX bacterial species (Brocadia sinica), sulfate-dependent ANAMMOX bacteria (Anammoxoglobus sulfate and Bacillus benzoevorans), and the highest nitrogen removal performance (74.3–76.7 kg-N/m3/d) in lab scale granule-based UASB reactors around the world were achieved. The characteristics, structure, packing pattern and floatation mechanism of the high-rate ANAMMOX granules in ANAMMOX reactors were also carefully illustrated by native researchers. Nowadays, some pilot and full-scale ANAMMOX reactors were constructed to treat different types of ammonium-rich wastewater including monosodium glutamate wastewater, pharmaceutical wastewater, and leachate. The prime objective of the present review is to elucidate the ongoing ANAMMOX research in China from lab scale to full scale applications, comparative analysis, and evaluation of significant findings and to set a design to usher ANAMMOX research in culmination.