Effect of light stress on lutein production with associated phosphorus removal from a secondary effluent by the autoflocculating microalgae consortium BR-UANL-01.

Microalgae have become promising microorganisms for generating high-value commercial products and removing pollutants in aquatic systems. This research evaluated the impact of sunlight intensity on intracellular pigment generation and phosphorus removal from secondary effluents by autoflocculating microalgae consortium BR-UANL-01 in photobioreactor culture. Microalgae were grown in a secondary effluent from a wastewater treatment plant, using a combination of low and high light conditions (photon irradiance; 44 µmol m-2 s-1 and ≈ 1270 µmol m-2 s-1, respectively) and 16:8 h light:dark and 24:0 h light:dark (subdivided into 18:6 LED:sunlight) photoperiods. The autoflocculant rate by consortium BR-UANL-01 was not affected by light intensity and achieved 98% in both treatments. Microalgae produced significantly more lutein, (2.91 mg g-1) under low light conditions. Phosphate removal by microalgae resulted above 85% from the secondary effluent, due to the fact that phosphorus is directly associated with metabolic and replication processes and the highest antioxidant activity was obtained in ABTS•+ assay by the biomass under low light condition (51.71% µmol ET g-1). In conclusion, the results showed that the autoflocculating microalgae consortium BR-UANL-01 is capable of synthesizing intracellular lutein, which presents antioxidant activity, using secondary effluents as a growth medium, without losing its autoflocculating activity and assimilating phosphorus.

Novel Thermotolerant Amylase from Bacillus licheniformis Strain LB04: Purification, Characterization and Agar-Agarose.

This study analyzed the thermostability and effect of calcium ions on the enzymatic activity of α-amylase produced by Bacillus licheniformis strain LB04 isolated from Espinazo Hot springs in Nuevo Leon, Mexico. The enzyme was immobilized by entrapment on agar-agarose beads, with an entrapment yield of 19.9%. The identification of the bacteria was carried out using 16s rDNA sequencing. The enzyme was purified through ion exchange chromatography (IEX) in a DEAE-Sephadex column, revealing a protein with a molecular weight of ≈130 kDa. The enzyme was stable at pH 3.0 and heat stable up to 80 °C. However, the optimum conditions were reached at 65 °C and pH 3.0, with a specific activity of 1851.7 U mg-1 ± 1.3. The agar-agarose immobilized α-amylase had a hydrolytic activity nearly 25% higher when compared to the free enzyme. This study provides critical information for the understanding of the enzymatic profile of B. licheniformis strain LB04 and the potential application of the microorganisms at an industrial level, specifically in the food industry.

Dissimilatory reduction of Fe(III) by a novel Serratia marcescens strain with special insight into the influence of prodigiosin.

A novel pigmented bacterium, initially identified as 11E, was isolated from a site historically known to have various iron-related ores. Phylogenetic analysis of this bacterial strain showed that it belongs to Serratia marcescens. This pigmented S. marcescens 11E cultured individually with glucose, acetate, and glycerol as electron donors along with the soluble electron acceptor iron (Fe) (III) citrate offered a large reduction extent (45.3 %, 31.4 %, and 13.5 %, respectively). On the other hand, when iron oxide (Fe2O3) is used as electron acceptor, the pigmented strain produced a null reduction extent. Surprisingly, the absence of prodigiosin on the bacterial surface (non-pigmented strain) resulted in a large reduction extent of the non-soluble iron form (20-49%). All these extents were comparable and, in some cases, superior to those presented in the literature. Additionally, in the present study, it was found that anthraquinone sulfonate (AQS) stimulated Fe(III) reduction of soluble and non-soluble Fe species only with pigmented S. marcescens. In contrast, in the culture media with the non-pigmented strain, the presence of AQS did not stimulate the Fe(III) reduction. These results suggest that the pigmented phenotype of S. marcescens 11E may perform non-soluble Fe(III) reduction by electron shuttling. In contrast, for the non-pigmented phenotype of this bacterium, non-soluble Fe(III) reduction seems to proceed by direct contact. Our study demonstrates that this bacterium may be used in bioreduction process of heavy metals or as a biocatalyst in bioelectrochemical devices.

Polyphosphate recovery by a native Bacillus cereus strain as a direct effect of glyphosate uptake.

Seven bacterial strains isolated from a glyphosate-exposed orange plantation site were exposed to 1 mM N-(phosphonomethyl)glycine supplied as a phosphorus source. While some exhibited good biodegradation profiles, the strain 6 P, identified as Bacillus cereus, was the only strain capable of releasing inorganic phosphate to the culture supernatant, while accumulating polyphosphate intracellularly along the experimentation time. The composition and purity of the intracellular polyphosphate accumulated by the strain 6 P were confirmed by FTIR analysis. To date, the biological conversion of glyphosate into polyphosphate has not been reported. However, given the importance of this biopolymer in the survival of microorganisms, it can be expected that this process could represent an important ecological advantage for the adaptation of this strain to an ecological niche exposed to this herbicide. The polyphosphate production yield was calculated as 4 mg l-1, while the glyphosate biodegradation kinetic constant was calculated on 0.003 h-1 using the modified Hockey-Stick first-order kinetic model, with a half-life of 279 h. Our results suggest that B. cereus 6 P is a potential candidate for the generation of an innovative biotechnological process to produce polyphosphate through the biodegradation of the herbicide glyphosate.

Antitumor activity of Chlorella sorokiniana and Scenedesmus sp. microalgae native of Nuevo León State, México.

Cancer cases result in 13% of all deaths worldwide. Unwanted side effects in patients under conventional treatments have led to the search for beneficial alternative therapies. Microalgae synthesize compounds with known in vitro and in vivo biological activity against different tumor cell lines. Therefore, native microalgae from the State of Nuevo Leon, Mexico may become a potential source of antitumor agents. The aim of the present study was to evaluate the in vitro cytotoxic effect of Nuevo Leon regional Chlorella sorokiniana (Chlorellales: Chlorellaceae) and Scenedesmus sp. (Chlorococcales: Scenedesmaceae). Native microalgae crude organic extracts cytotoxicity against murine L5178Y-R lymphoma cell line and normal lymphocyte proliferation were evaluated using the MTT reduction colorimetric assay. Cell death pathway was analyzed by acridine orange and ethidium bromide staining, DNA degradation in 2% agarose gel electrophoresis and caspases activity. Results indicated significant (p < 0.05) 61.89% ± 3.26% and 74.77% ± 1.84% tumor cytotoxicity by C. sorokiniana and Scenedesmus sp. methanol extracts, respectively, at 500 µg/mL, by the mechanism of apoptosis. This study contributes to Mexican microalgae biodiversity knowledge and their potential as antitumor agent sources.

Polishing of municipal secondary effluent using native microalgae consortia.

This work evaluates the use of native microalgae consortia for a dual role: polishing treatment of municipal wastewater effluents and microalgae biomass feedstock potential for biodiesel or biofertilizer production. An initial screening was undertaken to test N and P removal from secondary effluents and biomass production by 12 consortia. A subsequent treatment was performed by selected consortia (01 and 12) under three operational conditions: stirring (S), S + 12 h of daily aeration (S + A) and S + A enriched with CO2 (S + AC). All treatments resulted in compliance with environmental regulations (e.g. Directive 91/271/EEC) and high removal efficiency of nutrients: 64-79% and 80-94% of total N and PO43--P respectively. During the experiments it was shown that pH alkalinization due to microalgae growth benefits the chemical removal of ammonia and phosphorus. Moreover, advantages of pH increase could be accomplished by intermittent CO2 addition which in this research (treatment S + AC) promoted higher yield and lipid concentration. The resulting dry biomass analysis showed a low lipid content (0.5-4.3%) not ideal for biodiesel production. Moreover, the high rate of ash (29.3-53.0%) suggests that biomass could be readily recycled as a biofertilizer due to mineral supply and organic constituents formed by C, N and P (e.g. carbohydrate, protein, and lipids).

Metal-Induced Production of a Novel Bioadsorbent Exopolysaccharide in a Native Rhodotorula mucilaginosa from the Mexican Northeastern Region.

There is a current need to develop low-cost strategies to degrade and eliminate industrially used colorants discharged into the environment. Colorants discharged into natural water streams pose various threats, including: toxicity, degradation of aesthetics and inhibiting sunlight penetration into aquatic ecosystems. Dyes and colorants usually have complex aromatic molecular structures, which make them very stable and difficult to degrade and eliminate by conventional water treatment systems. The results in this work demonstrated that heavy metal-resistant Rhodotorula mucilaginosa strain UANL-001L isolated from the northeast region of Mexico produce an exopolysaccharide (EPS), during growth, which has colorant adsorption potential. The EPS produced was purified by precipitation and dialysis and was then physically and chemically characterized by Scanning Electron Microscopy, Fourier Transform Infrared Spectroscopy, and chemical elemental analysis. Here, the ability of the purified EPS produced to adsorb methylene blue (MB), which served as a model colorant, is studied. MB adsorption by the EPS is found to follow Langmuir Adsorption Isotherm kinetics at 25°C. Further, by calculating the Langmuir constant the adsorption capabilities of the EPS produced by the Rhodotorula mucilaginosa strain UANL-001L is compared to that of other adsorbents, both, microbially produced and from agroindustrial waste. The total adsorption capacity of the EPS, from the Rhodotorula mucilaginosa strain UANL-001L, was found to be two-fold greater than the best bioadsorbents reported in the literature. Finally, apart from determining which heavy metals stimulated EPS production in the strain, the optimal conditions of pH, heavy metal concentration, and rate of agitation of the growing culture for EPS production, was determined. The EPS reported here has the potential of aiding in the efficient removal of colorants both in water treatment plants and in situ in natural water streams.

Lipid production by pure and mixed cultures of Chlorella pyrenoidosa and Rhodotorula mucilaginosa isolated in Nuevo Leon, Mexico.

Given the well-known environmental drawbacks of using fossil fuels, advances in the field of alternative energy have become a worldwide technological priority. Special interest has been focused on the production of biodiesel obtained from oleaginous microorganisms. In the present research, lipid production by two species, microalgae Chlorella pyrenoidosa and yeast Rhodotorula mucilaginosa was assessed, independently and in mixed culture to evaluate a possible synergy. Fatty acid analysis was performed by gas chromatography. Among pure and mixed cultures of both strains and several culturing conditions, the highest biomass and lipid productivity was obtained by C. pyrenoidosa (8.05 and 1.62 g/L, respectively). The results of this study showed that both strains used are in fact oleaginous strains as they were found to reach up to 20 % of lipids, in addition, lipids in both pure and mixed cultures were mainly of triglycerides (>90 %), composed of fatty acid chains between 16 and 18 carbons.

Removal of chromium and lead by a sulfate-reducing consortium using peat moss as carbon source.

The effect of pre-treated peat moss on the ability of a sulfate-reducing microbial consortium to remove chromium and lead in solution was evaluated. The most active bacterial community (235.7 mmol H2S/g VSS) was selected from among eight consortia. The peat moss was pre-treated with different HCl concentrations and contact times. The best combination of treatments was 20% HCl for 10 min. The constant substrate affinity Ks was 740 mg COD/L and the ratio COD/SO4(2-) was 0.71. At pH 5, higher production of biogenic sulfide was observed. The up-flowpacked bed bioreactor operated at a flow of 8.3 mL/min for 180 h to obtain removal efficiency (by sulfate-reducing activity) of 90% lead and 65% chromium. It is important to consider that peat moss is a natural adsorbent that further influences the removal efficiency of metal ions.

Modelling sulphate-enhanced cadmium uptake by Zea mays from nutrient solution under conditions of constant free Cd2+ ion activity.

A controlled hydroponic experiment was undertaken to investigate Cd uptake in relation to the activity of Cd species in solution other than the free ion (Cd2+) by maintaining a constant Cd2+ activity under variable SO42- and Cl- concentrations exposed to maize (Zea mays var. Cameron) plants. The objectives of these experiments were: (1) to distinguish and quantify the different uptake rates of free and inorganic-complexed Cd from nutrient solution, and (2) to model the uptake of Cd by maize with a Biotic Ligand Model (BLM) in a system which facilitates the close examination of root characteristics. Results of the current experiments suggest that, in addition to the free ion, CdSO4(0) complexes are important factors in determining Cd uptake in nutrient solution by maize plants. Higher nominal SO4(2-) concentrations in solution generally resulted in a greater Cd accumulation by maize plants than predicted by the Cd2+ activity. A better integration of the complete dataset for the 3 harvest times (6, 9 and 11 days after treatment) was achieved by including consideration of both the duration of Cd exposure and especially the root surface area to express Cd uptake. Similarly, the fit of the BLM was also improved when taking into account exposure time and expressing uptake in terms of root morphological parameters.

The use of chloro-complexation to enhance cadmium uptake by Zea mays and Brassica juncea: testing a "free ion activity model" and implications for phytoremediation.

Maize and Indian mustard plants were studied to: 1) investigate the effect of Cl- complexation on Cd uptake from soil historically amended with sewage sludge (Cd 58 mg kg(-1)) and, 2) model the uptake of Cd by these plants with a Free Ion Activity Model (FIAM). Plants were treated with NaCl (50 to 300 mM in the soil pore water) along with controls using Na2SO4. Cadmium enhanced solubility in soil by Cl- generally reflected increases in Cd uptake by both plants. The free ion Cd2+ activity in soil solution, as modeled by WHAM-VI, remained almost unchanged despite the wide range of NaCl concentrations. Therefore, Na+ exchange for Cd2+ could not fully explain the differences in Cd content between the Cl- treatments because of the high buffering Cd2+ capacity in soiL Activities of Cd-chloro complexes showed the best correlations with the Cd concentrations in the plants compared to the activity of Cd2+. The FIAM showed a reasonable good fit for the plants when assuming competition by Cd2+ and CdCl+ for root sorption sites. Indirect evidence suggests that CaSO4 precipitation may have limited the formation of CdSO4 complexes and reduced Cd soil solubility. The implications of these results for phytoremediation are discussed.

Evaluating a 'biotic ligand model' applied to chloride-enhanced Cd uptake by Brassica juncea from nutrient solution at constant Cd2+ activity.

Evidence of chloride-enhanced cadmium uptake by plants in soil experiments has been reported. However, it is still unclear whether this finding is due to increased rates of Cd2+ diffusion to plant roots or the direct uptake of complexes such as CdC1+. A controlled hydroponic experiment was undertaken to distinguish and quantify the uptake rates of free and inorganic-complexed cadmium and to model the uptake of cadmium by Indian mustard plants with a 'biotic ligand model'. Plants were treated with NaCl (0 to 200 mM) including equivalent Na2SO4 treatments. Cadmium speciation in solution was calculated using the WHAM-VI model. Results of the current trials showed that higher Cl-concentrations in solution generally resulted in greater cadmium accumulation by plants than predicted by the Cd2+ activity. Activities of Cd-chloro complexes showed the best correlations with the cadmium concentrations in the plants compared with the activity of Cd2+. The biotic ligand model showed a reasonable good fit for the plants when assuming competition by Cd2+ and CdCl+ for sorption sites at root level. The relative values of the two reaction constants suggest that root affinity for Cd2+ is 3.4 times greater than for CdCl+. Nevertheless this clearly indicates a substantial role for chloro-complexed cadmium accumulation.

Plant screening of halophyte species for cadmium phytoremediation.

Chloride (Cl-) has been related to increased phytoavailability of cadmium (Cd) in soil. A glasshouse experiment using a historically contaminated soil was undertaken to evaluate the effect of chloride on Cd uptake by salt-tolerant plants and possibly quantify the uptake of discrete Cd species (e.g. Cd2+, CdCl+, CdCl(0)2). Chloride treatments were applied as 100 mM NaCl and compared with equivalent Na2SO4 treatments. Activities of Cd species in soil pore water were calculated using the WHAM-VI speciation model. Cadmium solubility and uptake by plants was generally enhanced by addition of chloride to soil. Good correlations were found between Cd uptake and concentrations of Cd chloride complexes in soil pore water.