Xuwang Zhang

Performance and microbial community dynamics in bioaugmented aerated filter reactor treating with coking wastewater

In this study, zeolite-biological aerated filters (Z-BAFs) bioaugmented by free and magnetically immobilized cells of Arthrobacter sp. W1 were designed to treat coking wastewater containing high concentrations of phenol and naphthalene along with carbazole (CA), dibenzofuran (DBF), and dibenzothiophene (DBT). All treatments were carried out for a period of 100days and the data indicated that bioaugmented Z-BAFs with magnetically immobilized cells was most efficient for treating coking wastewaters. Illumina high-throughput sequencing was used to reveal the microbial community structures of Z-BAFs. Both bioaugmentation treatments could accelerate the shift of the bacterial community structures. The introduced strain W1 remained dominant in the bioaugmented Z-BAFs with magnetically immobilized cells, indicating both strain W1 and the indigenous degrading bacteria played the most significant role in the treatment. Overall, bioaugmented Z-BAF with magnetically immobilized cells can be used to efficiently degrade phenol, naphthalene, CA, DBF, and DBT in coking wastewater.

Aerobic decolorization and degradation of azo dyes by growing cells of a newly isolated yeast Candida tropicalis TL-F1

The aim of this work was to investigate the decolorization and degradation of azo dyes by growing cells of a new yeast strain TL-F1 which was isolated from the sea mud. Strain TL-F1 was identified as Candida tropicalis on the basis of 28S rDNA analysis. Various azo dyes (20mg/L) were efficiently decolorized through aerobic degradation. Meantime, the effects of different parameters on both decolorization of Acid Brilliant Scarlet GR and growth of strain TL-F1 were investigated. Furthermore, possible degradation pathway of the dye GR was proposed through analysis of metabolic products using UV-Vis spectroscopy and HPLC-MS methods. As far as it is known, it is the first systematic research on a C. tropicalis strain which is capable of efficiently decolorizing various azo dyes under aerobic condition. This work provides a potentially useful microbial strain TL-F1 for treatment of azo dye contaminated wastewater.

Aerobic decolorization and degradation of Acid Red B by a newly isolated Pichia sp. TCL.

A yeast strain with strong abilities to decolorize various azo dyes aerobically was isolated from the sea mud. The strain designated as TCL was identified as Pichia sp. on the basis of 18S rDNA analysis. More than 90% of Acid Red B (100mg/L) was decolorized within 10h in the Martin Broth at 30°C and 150r/min, and strain TCL could tolerate up to 1000mg/L of the dye. Meantime, the effects of different physicochemical parameters (media, concentrations of glucose, NH(4)Cl, initial dye and NaCl) were investigated to improve the removal efficiency. The significant biodegradation process of Acid Red B rather than inactive surface adsorption was confirmed by UV-vis, HPLC analysis and colorless microbial cells. In addition, the metabolic products and partial degradation pathway were proposed with the help of HPLC-MS analysis. To the best of our knowledge, it is the first time that a yeast strain of Pichia sp. has been reported with the excellent decolorizing ability against azo dyes under shaking conditions. This work conferred the utilization possibility of strain TCL in the biological treatment of dyeing wastewater.

Responses of Microbial Communities to Single-Walled Carbon Nanotubes in Phenol Wastewater Treatment Systems

The expanding use of single-walled carbon nanotubes (SWCNTs) raises environmental concerns. Wastewater treatment systems are potential recipients of SWCNTs containing influent, yet the impacts of SWCNTs on these systems are poorly documented. In this study, the microbial responses to SWCNTs in simulated phenol wastewater treatment systems were investigated. The phenol removal rates were improved in all SWCNTs-treated sequencing batch reactors during the first 20 days, but when facing higher phenol concentration (1000 mg/L) after 60 days, reactors with the highest concentration (3.5 g/L) of SWCNTs exhibited a notably decreased phenol removal capacity. Cell viability tests, scanning electron microscopy analysis and DNA leakage data suggested that SWCNTs protected microbes from inactivation, possibly by producing more bound extracellular polymeric substances (EPS), which could create a protective barrier for the microbes. Illumina sequencing of 16S rRNA gene amplicons revealed that the bacterial diversity did not change significantly except for a minor reduction after the immediate addition of SWCNTs. Bacterial community structure significantly shifted after SWCNTs addition and did not recover afterward. Zoogloea increased significantly upon SWCNTs shocking. At the final stage, Rudaea and Mobilicoccus increased, while Burkholderia, Singulisphaera, Labrys and Mucilaginibacter decreased notably. The shifts of these dominant genera may be associated with altered sludge settling, aromatic degradation and EPS production. This study suggested that SWCNTs exerted protective rather than cytotoxic effects on sludge microbes of phenol wastewater treatment systems and they affected the bacterial community structure and diversity at test concentrations. These findings provide new insights into our understanding of the potential effects of SWCNTs on wastewater treatment processes.

Illumina MiSeq Sequencing Reveals Diverse Microbial Communities of Activated Sludge Systems Stimulated by Different Aromatics for Indigo Biosynthesis from Indole.

Indole, as a typical N-heteroaromatic compound existed in coking wastewater, can be used for bio-indigo production. The microbial production of indigo from indole has been widely reported during the last decades using culture-dependent methods, but few studies have been carried out by microbial communities. Herein, three activated sludge systems stimulated by different aromatics, i.e. naphthalene plus indole (G1), phenol plus indole (G2) and indole only (G3), were constructed for indigo production from indole. During the operation, G1 produced the highest indigo yield in the early stage, but it switched to G3 in the late stage. Based on LC-MS analysis, indigo was the major product in G1 and G3, while the purple product 2-(7-oxo-1H-indol-6(7H)-ylidene) indolin-3-one was dominant in G2. Illumina MiSeq sequencing of 16S rRNA gene amplicons was applied to analyze the microbial community structure and composition. Detrended correspondence analysis (DCA) and dissimilarity tests showed that the overall community structures of three groups changed significantly during the operation (P<0.05). Nevertheless, the bacteria assigned to phylum Proteobacteria, family Comamonadaceae, and genera Diaphorobacter, Comamonas and Aquamicrobium were commonly shared dominant populations. Pearson correlations were calculated to discern the relationship between microbial communities and indigo yields. The typical indigo-producing populations Comamonas and Pseudomonas showed no positive correlations with indigo yields, while there emerged many other genera that exhibited positive relationships, such as Aquamicrobium, Truepera and Pusillimonas, which had not been reported for indigo production previously. The present study should provide new insights into indigo bio-production by microbial communities from indole.

Influence and optimization of growth substrates on indigo formation by a novel isolate Acinetobacter sp. PP-2.

In this study, a novel indigo-producing bacterial strain PP-2 was isolated from activated sludge. It was identified as Acinetobacter sp. according to phylogenetic similarity of 16S rRNA gene sequence. This isolate was able to produce indigo from indole by utilizing a wide range of aromatic hydrocarbons. The results of SDS-PAGE analysis showed that the enzyme system induced by phenol was more abundant than that induced by other aromatic hydrocarbons. And the effects of metal ions on indigo production were also investigated, which indicated that the activity of cells induced with phenol could be inhibited by 0.5mM Fe(3+). Response surface methodology (RSM) was applied to optimize the process of indigo bio-production. The results exhibited that the maximal yield was achieved with 157.92 mg L(-1) phenol and 205.32 mg L(-1) indole. Under the optimal conditions, the indigo yield and transformation efficiency of indole were 202.92 mg L(-1) and 88%, respectively.

Optimization of indigo production by a newly isolated Pseudomonas sp. QM

Optimization of indigo production process from indole using a newly isolated phenol‐degrading bacterial strain was performed by Plackett‐Burman design and response surface methodology. The strain designated as QM was identified as Pseudomonas sp. according to 16S rDNA analysis. Spectrum analysis of indole biotransformation products revealed the presence of indigo and a by‐product indirubin. To improve indigo yield, Plackett‐Burman design was used to select significant factors from 8 viriables. Then response surface methodology based on a 23 central composite design was used to further optimize the transformation process. Under the optimal conditons, strain QM can produce 27.20 mg/l indigo after 24 h cultivation at 30 °C, which was 151.3% higher than that from the initial conversion condition. The results indicated that Pseudomonas sp. QM should be a potential candidate for indigo industial production. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Characterization of a Novel Phenol Hydroxylase in Indoles Biotranformation from a Strain Arthrobacter sp. W1

Background Indigoids, as popular dyes, can be produced by microbial strains or enzymes catalysis. However, the new valuable products with their transformation mechanisms, especially inter-conversion among the intermediates and products have not been clearly identified yet. Therefore, it is necessary to investigate novel microbial catalytic processes for indigoids production systematically. Findings A phenol hydroxylase gene cluster (4,606 bp) from Arthrobacter sp. W1 (PHw1) was obtained. This cluster contains six components in the order of KLMNOP, which exhibit relatively low sequence identities (37–72%) with known genes. It was suggested that indole and all the tested indole derivatives except for 3-methylindole were transformed to various substituted indigoid pigments, and the predominant color products derived from indoles were identified by spectrum analysis. One new purple product from indole, 2-(7-oxo-1H-indol-6(7H)-ylidene) indolin-3-one, should be proposed as the dimerization of isatin and 7-hydroxylindole at the C-2 and C-6 positions. Tunnel entrance and docking studies were used to predict the important amino acids for indoles biotransformation, which were further proved by site-directed mutagenesis. Conclusions/Significance We showed that the phenol hydroxylase from genus Arthrobacter could transform indoles to indigoids with new chemical compounds being produced. Our work should show high insights into understanding the mechanism of indigoids bio-production.

Systematic investigation and microbial community profile of indole degradation processes in two aerobic activated sludge systems

Indole is widely spread in various environmental matrices. Indole degradation by bacteria has been reported previously, whereas its degradation processes driven by aerobic microbial community were as-yet unexplored. Herein, eight sequencing batch bioreactors fed with municipal and coking activated sludges were constructed for aerobic treatment of indole. The whole operation processes contained three stages, i.e. stage I, glucose and indole as carbon sources; stage II, indole as carbon source; and stage III, indole as carbon and nitrogen source. Indole could be completely removed in both systems. Illumina sequencing revealed that alpha diversity was reduced after indole treatment and microbial communities were significantly distinct among the three stages. At genus level, Azorcus and Thauera were dominant species in stage I in both systems, while Alcaligenes, Comamonas and Pseudomonas were the core genera in stage II and III in municipal sludge system, Alcaligenes and Burkholderia in coking sludge system. In addition, four strains belonged to genera Comamonas, Burkholderia and Xenophilus were isolated using indole as sole carbon source. Burkholderia sp. IDO3 could remove 100 mg/L indole completely within 14 h, the highest degradation rate to date. These findings provide novel information and enrich our understanding of indole aerobic degradation processes.

Extradiol dioxygenase–SiO2 sol–gel modified electrode for catechol and its derivatives detection

A feasible and sensitive biosensor for catechol and its derivatives using 2,3-dihydroxybiphenyl 1,2-dioxygenase (BphC)-modified glassy carbon electrode was successfully constructed by polyvinyl alcohol-modified SiO₂ sol-gel method. The as-prepared biosensor was characterized by electrochemical impedance spectroscopy, and the surface topography of the film was imaged by atomic force microscope. Liquid chromatography-tandem mass spectrometry was applied to reveal the catalytic mechanism. BphC embedded in SiO₂ gel maintained its bioactivity well and exhibited excellent eletrocatalytical response to both catechol and some of its derivatives (such as 3-methylcatechol and 4-methylcatechol). The biosensor showed a linear amperometric response range between 0.002 mM and 0.8 mM catechol. And the sensitivity was 1.268 mA/(mM cm²) with a detection limit of 0.428 μM for catechol (S/N = 3). Furthermore, the BphC biosensor exhibited perfect selectivity for catechol in the mixtures of catechol and phenol. It was suggested that this flexible protocol would open up a new avenue for designing other ring-cleavage enzyme biosensors, which could be widely used for monitoring various kinds of environmental pollutants.