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Information on the catalytic role of mlrA gene-encoded enzyme (MlrA) in microcystin-RR (MC-RR) biodegradation was limited. This study succeeded in expressing mlrA homolog of Novosphingobium sp. THN1 in heterologous host for the first time, by constructing a recombinant bacterium. Mass spectrometric analysis showed that the recombinant MlrA hydrolyzed MC-RR into linear intermediate product by cleaving the peptide bond between Adda and arginine residue, greatly detoxifying MC-RR. This finding clearly manifested that the MlrA homolog of THN1 strain possesses its original catalytic function, and ring-opening constituted the first step in MC-RR biodegradation pathway of THN1 strain. Moreover, MC-RR degradation by intact recombinant cells and cell-free crude enzyme (CE) from recombinant was compared. Results exhibited that intact recombinant was able to degrade 20 μg mL −1 MC-RR more quickly than CE, with the maximum rate of 9.22 μg mL −1 h −1 in the first 8 h. Thus, this study provided new insights on the catalytic activity and roles of MlrA originated from THN1 strain in MC-RR biodegradation process, which lay a foundation for efficiently removing and detoxifying MC-RR, and exploring downstream steps in MC-RR biodegradation pathway of THN1 strain. [ABSTRACT FROM AUTHOR]

Cyanobacterial bloom, a common phenomenon nowadays often results in the depletion of dissolved oxygen (hypoxia) and releases microcystin-RR (MC-RR) in the water. Information on the combined effects of MC-RR and hypoxia on the goldfish is lacking, therefore, this study is aimed at evaluating the effect of two doses of MC-RR on the antioxidants and globin mRNA of goldfish under normoxia, hypoxia and reoxygenation. The result showed that MC-RR at both doses (50 and 200μgkg−1 body weight) significantly (p<0.05) induced superoxide dismutase activities in the liver and kidney but catalase activities and total antioxidant capacity were low in these organs during hypoxia and reoxygenation compared to normoxia and control. Myoglobin and neuroglobin mRNAs in MC-RR group were significantly induced in the brain only and are believed to protect the brain from oxidative damage. However, other organs were unprotected and extensive damage was observed in the liver cells. Our results clearly demonstrated that MC-RR and hypoxia-reoxygenation transitions were synergistically harmful to the goldfish and could impair its adaptation to hypoxia, especially during reoxygenation. [ABSTRACT FROM AUTHOR]

Abstract: 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, 1,008bp) 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 40mgL−1 was completely biodegraded under total protein of 350mgL−1 within 0.25h. 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. [Copyright &y& Elsevier]

Abstract: Tobacco BY-2 cells were exposed to microcystin-RR (MC-RR) at two concentrations, 60μgmL−1 and 120μgmL−1, to study the changes in morphology and ultrastructure of cells as a result of the exposure. Exposure to the lower concentration for 5d led to typical apoptotic morphological changes including condensation of nuclear chromatin, creation of a characteristic ‘half moon’ structure, and cytoplasm shrinkage and decreased cell volume, as revealed through light microscopy, fluorescence microscopy, and transmission electron microscopy, respectively. Exposure to the higher concentration, on the other hand, led to morphological and ultrastructural changes typical of necrosis, such as rupture of the plasma membrane and the nuclear membrane and a marked swelling of cells. The presence of many vacuoles containing unusual deposits points to the involvement of vacuoles in detoxifying MC-RR. Results of the present study indicate that exposure of tobacco BY-2 cells to MC-RR at a lower concentration (60μgmL−1) results in apoptosis and that to a higher concentration (120μgmL−1), in necrosis. [Copyright &y& Elsevier]

Abstract: The production of cyanobacterial toxins microcystins (MCs) by cyanobacterial bloom which may promote the growth of tumor in human liver is a growing environmental problem worldwide. In this paper, the adsorption of MC-RR and LR, which were extracted from cyanobacterial cells in Dianchi Lake in China, by carbon nanotubes (CNTs), wood-based activated carbon (ACs) and clays were investigated. Compared with ACs and clay materials of sepiolite, kaolinite and talc tested, CNTs were found to have a strong ability in the adsorption of MCs. At the concentrations of 21.5mgl−1 MC-RR and 9.6mgl−1 MC-LR in 50mmol phosphate buffer solution (pH 7.0), the adsorption amounts of MCs by CNTs with the range of outside diameter from 2 to 10nm were 14.8 and 5.9mgg−1, which were about four times higher than those by other adsorbents tested. It was shown that with the decrease of CNTs outside diameters from 60 to 2nm, the adsorption amount of MCs was apparently increased, however the size of CNTs particles formed in solution declined. This result implies that the size of CNTs tube pore that is fit for the molecular dimension of MCs plays a dominant role. Furthermore the specific surface area of CNTs was also found to be a factor in the adsorption of MCs. The results suggested that the selection of suitable size of CNTs as a kind of adsorbent is very important in the efficient eliminating MCs from drinking water in future. [Copyright &y& Elsevier]

Abstract: A new and efficient method for the degradation of microcystins (one family of blue algal toxins) was developed and studied. Microcystins (MCs) in water were directly and effectively removed by active chlorine transformed in situ from the naturally existing Cl− in water resource using electrochemical method. Titanium coated with RuO2 and TiO2 was used as the anode. Microcystin-RR (MCRR) and Microcystin-LR (MCLR) were chosen as the model compounds of MCs. The results suggested that 20.87mgl−1 MCs (12.58mgl−1 MCRR and 8.29mgl−1 MCLR) in aqueous solution with 1.85mM Cl− could be synchronously decomposed within 15min electrolysis under the condition of the current density 8.89mAcm−2, 20°C and pH 7.00. The qualitative analysis showed that the heptapetide ring and the Adda group of both treated MCs were changed. The results also indicated that the removal rates of both MCs increased with the increasing of chloride concentration and applied current density, but decreased with the increasing of initial concentration of MCs and initial pH of electrolyte. In the absence of Cl−, only a small fraction of both MCs were decomposed by direct anodic oxidation, while their almost complete removals could be obtained in the case of indirect electrooxidation with in situ electrogenerated active chlorine from Cl− in water. [Copyright &y& Elsevier]