Hai Yan

Toxicity and bioaccumulation of copper in three green microalgal species

The effective concentrations of copper on the inhibition of the growth of Scenedesmus obliquus, Chlorella pyrenoidosa and Closterium lunula at 96 h (96 h EC50) were determined to be 50, 68 and 200 microg/l, respectively. The low initial bioaccumulation of Cu by C. lunula was found to be responsible for its tolerance to Cu. The amount of Cu accumulated by all three microalgae reached the maximum value and decreased quickly after the peak followed by a slow decrease over the next 6 d. Bioaccumulation of Cu by C. lunula was directly proportional to the initial Cu concentration. After reaching the first peak after 1 d, the bioconcentration factor of Cu by microalgae declined to its minimum value during the exponential growth phase but increased in the stationary growth phase again. This indicates that desorption of Cu from microalgae was higher during the exponential growth phase but lower in the stationary growth phase. Smaller microalgae with low 96 h EC50 values are more efficient in removing Cu from wastewater.

Effective removal of microcystins using carbon nanotubes embedded with bacteria

Adsorption of microcystins (MCs) by carbon nanotubes (CNTs) and clay materials was studied. Compared with various clays tested, CNTs showed a much stronger ability to adsorb MC-RR and LR that were two typical types of microcystins found in China. At initial 21.0 mg/L of MC-RR and 9.5 mg/L of MC-LR in solution, the adsorption amounts of MC-RR and LR by CNTs were 14.8 and 6.7 mg/g that were about five times higher than those by the clay materials of sepiolite, kaolinite and talc, etc. In the presence of CNTs and the bacterialRalstonia solanacearum that was firstly isolated and used for the biodegradation of MCs by the authors, a remarkable removal of MCs from water were observed. The mechanism was that CNTs could absorb large amount of both MCs and the embeddedR. solanacearum so that, even when diluted by a large amount of water, the concentrations of both organic pollutants and the added bacteria could be largely enhanced on the surface of CNTs where a concerted biodegradation reaction was effectively conducted. This finding could be important for the further development of practical techniques to eliminate MCs from polluted drinking waters.

Biodegradation of methyl parathion by Acinetobacter radioresistens USTB-04

Biodegradation of methyl parathion (MP), a widely used organophosphorus pesticide, was investigated using a newly isolated bacterium strain Acinetobacter radioresistens USTB-04. MP at an initial concentration of 1200 mg/L could be totally biodegraded by A. radioresistens USTB-04 as the sole carbon source less than 4 d in the presence of phosphate and urea as phosphorus and nitrogen sources, respectively. Biodegradation of MP was also achieved using cell-free extract of A. radioresistens USTB-04. MP at an initial concentration of 130 mg/L was completely biodegraded in 2 h in the presence of cell-free extract with a protein concentration of 148.0 mg/L, which was increased with the increase of pH from 5.0 to 8.0. Contrary to published reports, no intermediate or final degradation metabolites of MP could be observed. Thus we suggest that the cleavage of C-C bond on the benzene ring other than P-O bond may be the biodegradation pathway of MP by A. radioresistens USTB-04.

Biodegradation of Aniline by a Newly Isolated Delftia sp. XYJ6

Abstract A promising gram-negative bacterial strain for the biodegradation of aniline as the sole carbon, nitrogen and energy sources was successfully isolated and identified as Delftia sp. XYJ6. The optimal temperature and pH for both the growth of Delftia sp. XYJ6 and the biodegradation of aniline were 30°C and 7.0, respectively. Initial aniline of 2000 mg·L −1 could be completely removed by the strain at 22 h, which showed that Delftia sp. XYJ6 had a strong ability in the biodegradation of aniline. It indicated that aniline was firstly converted to catechol catalyzed by aniline dioxygenase as a first product, which was then further biodegraded to cis, cis -muconic acid catalyzed by the catechol 1,2-dioxygenase of Delftia sp. XYJ6 as a second product. Cis, cis -muconic acid could also be further biodegraded to other small compound again. The pathway for the biodegradation of aniline by Delftia sp. XYJ6 was not previously reported.

Microbial biodegradation of microcystin-RR by bacterium Sphingopyxis sp. USTB-05

A strain, USTB-05, isolated from Lake Dianchi, China, degraded the cyanobacterial toxin microcystin-RR (MC-RR) at the rate of 16.7 mg/L per day. Analysis of 16S rDNA sequence showed that the strain was Sphingopyxis sp. Enzymatic degradation pathways for MC-RR by Sphingopyxis sp. USTB-05 were identified. Adda-Arg peptide bond of MC-RR was cleaved and then a hydrogen and a hydroxyl were combined onto the NH2 group of Adda and the carboxyl group of arginine to form a linear molecule as intermediate product within the first few hours. Then, through dehydration reaction, two hydrogen of amino group on arginine reacted with adjacent hydroxyl on carbon to form a linear MC-RR with two small peptide rings as the final product after 24 hr. These biodegradation pathways were different from those reported for other strains, implying that MC-RR may undergo different transformations and different products were formed due to various bacteria in natural lakes and reservoirs.

Biodegradation of Microcystin-RR by a New Isolated Sphingopyxis sp. USTB-05

Abstract A promising bacterial strain for the biodegradation of Microcystins (MCs) was isolated from Dianchi lake in China and identified as Sphingopyxis sp. USTB-05 by the analysis of 16s rDNA. Initial MC-RR of 42.3 mg·L −1 was completely degraded by USTB-05 within 36 h, which was a relatively high biodegradation rate of MC-RR. With the cell-free extract (CE) of Sphingopyxis sp. USTB-05, MC-RR was biodegraded at a more rapid biodegradation rate compared with its strain, so that initial MC-RR of 42.3 mg·L −1 was completely biodegraded within 10 h. During the bio-reaction of MC-RR catalyzed by CE, two intermediate metabolites and a dead-end product of MC-RR were observed on HPLC profiles and all of them had similar scanning profiles in the wavelength from 200 to 300 nm, indicating that the group of Adda in all products of MC-RR remained intact.

Characterization of the first step involved in enzymatic pathway for microcystin-RR biodegraded by Sphingopyxis sp. USTB-05

Enzymes encoded by genes biodegrading microcystins (MCs) can help reveal the function of genes and biodegradation pathway of MCs. Here the first and important gene (USTB-05-A, 1,008 bp) involved in biodegradation of microcystin-RR (MC-RR) was cloned from Sphingopyxis sp. USTB-05 and firstly expressed in Escherichia coli BL21 (DE3) with an expression vector of pGEX4T-1 successfully. The nucleotide sequences of cloned USTB-05-A possessed 92.5% homology to that of mlA reported in Sphingomonas sp. strain ACM-3962. The deduced amino acid sequences containing the cleavage sites of 26th (alanine) and 27th (leucine) showed 83% identical to that of MlrA. The cell-free extract (CE) of recombinant E. coli BL21 (DE3) containing USTB-05-A had high activity for biodegrading MC-RR. Initial MC-RR of 40 mg L(-1) was completely biodegraded under total protein of 350 mg L(-1) within 0.25 h. A product derived from MC-RR appeared distinctly with the decrease of MC-RR peak on the profile of HPLC. The product (m/z 1056.5) had molecular weight of 18 higher than that of MC-RR (m/z 1038.7). The findings provided the positive evidences that biodegradation of MC-RR began with the breakage of cyclic MC-RR and then it was converted to linear MC-RR as the first product catalyzed by first enzyme of Sphingopyxis sp. USTB-05.

Effect of Bacillus subtilis CGMCC 1.1086 on the growth performance and intestinal microbiota of broilers

Probiotics have been proved to be the most preferred and effective alternative to antibiotics as growth promoter and pathogens inhibitor in poultry industry. In this study Bacillus subtilis CGMCC 1.1086 as a probiotic bacterium was administered in diet and its effects on both the growth performance and the caecal microbiota of broilers were evaluated.

Effects of nitrogen forms on the production of cyanobacterial toxin microcystin-LR by an isolated Microcystis aeruginosa

Abstract A cyanobacterial strain, which produced high content of microcystin-LR (MC-LR) but no microcystin-RR (MC-RR), was isolated from the hypertrophic Dianchi Lake in China and identified as Microcystis aeruginosa DC-1. Effects of nitrogen containing chemicals and trace elements on the growth and the production of MC-LR by this strain were studied. In the presence of bicine, compared with urea and ammonium, nitrate greatly promoted the growth and the production of MC-LR. However, leucine and arginine, which were the constitutional components in the molecular structure of MC-LR or RR, inhibited the production of MC-LR. Iron and silicon up to 10 mg/L had little effects on the growth of M. aeruginosa DC-1, but the production of MC-LR was apparently enhanced. Under all conditions studied here, only MC-LR but no RR was detected within the cells of M. aeruginosa DC-1. Thus, chemical forms of nitrogen, rather than the usually concerned the total nitrogen, and trace elements played important roles in the production of MC toxins during cyanobacterial blooms.

Cloning and expression of the first gene for biodegrading microcystin LR by Sphingopyxis sp. USTB-05

Harmful cyanobacterial blooms are a growing environmental problem worldwide in natural waters, the biodegradation is found to be the most efficient method for removing microcystins (MCs) produced by harmful cyanobacteria. Based on the isolation of a promising bacterial strain of Sphingopyxis sp. USTB-05 for biodegrading MCs, we for the first time cloned and expressed a gene USTB-05-A (HM245411) that is responsible for the first step in the biodegradation of microcystin LR (MC-LR) in E. coli DH5alpha, with a cloning vector of pGEM-T easy and an expression vector of pGEX-4T-1, respectively. The cell-free extracts (CE) of recombinant E. coli DH5alpha containing USTB-05-A had high activity for biodegrading MC-LR. The initial MC-LR concentration of 40 mg/L was completely biodegraded within 1 hr in the presence of CE with a protein concentration of 0.35 mg/mL. Based on an analysis of the liquid chromatogram-mass spectrum (LC-MS), the enzyme encoded by gene USTB-OS-A was found to be active in cleaving the target peptide bond between 3-amino-9-methoxy-2,6, 8-trimethyl-10-phenyl-deca-4,6-dienoic acid (Adda) and arginine of MC-LR, and converting cyclic MC-LR to linear MC-LR as a first product that is much less toxic than parent MC-LR, which offered direct evidence for the first step on the pathway of MC-LR biodegradation by Sphingopyxis sp. USTB-05.