RESUMEN
We have recently shown that SmbP, the small metal-binding protein of Nitrosomonas europaea, can be employed as a fusion protein to express and purify recombinant proteins and peptides in Escherichia coli. SmbP increases solubility, allows simple, one-step purification through affinity chromatography, and provides superior final yields due to its low molecular weight. In this work, we report for the first time the use of SmbP to produce a recombinant peptide with anticancer activity: the antitumor-analgesic peptide (BmK-AGAP), a neurotoxin isolated from the venom of the Chinese scorpion Buthus martensii Karsch. This peptide was expressed in Escherichia coli SHuffle for correct, cytoplasmic, disulfide bond formation and tagged with SmbP at the N-terminus to improve its solubility and allow purification using immobilized metal affinity chromatography. SmbP_BmK-AGAP was found in the soluble fraction of the cell lysate. After purification and removal of SmbP by digestion with enterokinase, 1.8 mg of pure and highly active rBmK-AGAP was obtained per liter of cell culture. rBmK-AGAP exhibited antiproliferative activity on the MCF-7 cancer cell line, with a half-maximal inhibitory concentration value of 7.24 µM. Based on these results, we considered SmbP to be a suitable carrier protein for the production of recombinant, biologically active BmK-AGAP. We propose that SmbP should be an attractive fusion protein for the expression and purification of additional recombinant proteins or peptides that display anticancer activities.
RESUMEN
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.
Asunto(s)
Compuestos Férricos/metabolismo , Prodigiosina/metabolismo , Serratia marcescens , Enzimas , Filogenia , ARN Ribosómico 16S/genética , Serratia marcescens/genética , Serratia marcescens/aislamiento & purificación , Serratia marcescens/metabolismoRESUMEN
The capacity of Cladosporium cladosporioides biomass for removal of Cr(VI) in aqueous solutions was evaluated. A 2 × 2 factorial experiment design was used to study the effects of pH and biomass doses. Lower pH values and larger biomass doses increased the capacity of C. cladosporioides biomass for removal of Cr(VI), reaching a reduction capacity of 492.85 mg g-1, a significantly higher value compared to other biomass reported. Cr(VI) removal kinetic rates followed a pseudo-second order model, like other fungal biomass reported previously. The apparent adsorption process was described well by the Freundlich isothermal model. However, determination of total chromium indicated that adsorption of Cr(VI) was followed by a redox reaction that released proportional quantities of Cr(III) into the experimental supernatant, suggesting a parallel adsorption-reduction process. Comparison of Fourier transform infrared spectroscopy spectra of C. cladosporioides biomass before and after the reduction process demonstrated the involvement of positively charged amino groups in the Cr(VI) adsorption-reduction process.
Asunto(s)
Cromo/metabolismo , Cladosporium/metabolismo , Contaminantes Químicos del Agua/metabolismo , Adsorción , Biodegradación Ambiental , Biomasa , Cladosporium/crecimiento & desarrollo , Concentración de Iones de Hidrógeno , Cinética , Oxidación-Reducción , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
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 ABTSâ¢+ 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.
RESUMEN
Cellulose, the most abundant biopolymer on earth, is produced at different ratios by all land plants. Since the morphology and crystallinity of cellulose are key factors involved in its enzymatic hydrolysis, in the present work, we tackled the study of the effects of such variables on the nanocellulose conversion into glucose. Cellulase from Trichoderma sp at 37 °C was used to produce glucose, the best results were found for the cellulose nanoplatelets (S-CNP) after 60 h of hydrolysis, which afforded a conversion of 47% to glucose, in contrast to 15% for the non-purified sample (W-CP) and 22% for microcrystalline cellulose (MCC20) used as control. The X-ray diffractogram recorded on the samples showed an initial crystallinity index of 45%, 54% and 72% for W-CNP, S-CNP and MCC20, respectively. Also, we showed that after 24 h of hydrolysis, long cellulose nanofibrils (∅ ≈ 30 nm) were found as a residue.
Asunto(s)
Celulasa/metabolismo , Celulosa/química , Nanoestructuras/química , Hidrólisis , Temperatura , Trichoderma/enzimologíaRESUMEN
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.