RESUMEN
In the present study, a new extracellular polysaccharide (EPS-M816) was obtained during the growth of Mesorhizobium loti Semia 816 on a crude glycerol-based medium. EPS-M816 precipitate mainly consisted of carbohydrates (82.54%) and proteins (11.31%), and the weight average molecular weight was estimated at 1.646â¯×â¯106â¯Da. The biopolymer was characterized by FT-IR and NMR spectroscopy, and was found to have typical functional groups of other rhizobial polysaccharides. Furthermore, the rheological and emulsifying properties were investigated. The EPS-M816 solution (1.0% w/v) showed typical pseudoplastic non-Newtonian fluid behavior, and the addition of sodium and potassium chloride (1â¯molâ¯L-1) increased the apparent viscosity. Regarding its emulsification activity, EPS-M816 formed emulsions with different food-grade vegetable oils (soybean, rice, canola, sunflower and corn oils), showing emulsification index values over 65% in 24â¯h, indicative of strong emulsion-stabilizing capacity. The biopolymer was able to form gels with texture parameters similar to those reported for xanthan gum and low syneresis. Overall, these results suggest that EPS-M816 is a good candidate for application in the food, cosmetics and pharmaceutical industries as a thickening, gelling, stabilizing and emulsifying agent.
Asunto(s)
Medios de Cultivo/química , Emulsionantes/metabolismo , Glicerol/farmacología , Mesorhizobium/crecimiento & desarrollo , Mesorhizobium/metabolismo , Polisacáridos Bacterianos/biosíntesis , Reología , Emulsionantes/química , Mesorhizobium/efectos de los fármacos , Peso Molecular , Polisacáridos Bacterianos/químicaRESUMEN
The aminopeptidase gene from Mesorhizobium SEMIA3007 was cloned and overexpressed in Escherichia coli. The enzyme called MesoAmp exhibited optimum activity at pH 8.5 and 45 °C and was strongly activated by Co2+ and Mn2+. Under these reaction conditions, the enzyme displayed Km and kcat values of 0.2364 ± 0.018 mM and 712.1 ± 88.12 s-1, respectively. Additionally, the enzyme showed remarkable stability in organic solvents and was active at high concentrations of NaCl, suggesting that the enzyme might be suitable for use in biotechnology. MesoAmp is responsible for 40% of the organism's aminopeptidase activity. However, the enzyme's absence does not affect bacterial growth in synthetic broth, although it interfered with biofilm synthesis and osmoregulation. To the best of our knowledge, this report describes the first detailed characterization of aminopeptidase from Mesorhizobium and suggests its importance in biofilm formation and osmotic stress tolerance. In summary, this work lays the foundation for potential biotechnological applications and/or the development of environmentally friendly technologies and describes the first solvent- and halo-tolerant aminopeptidases identified from the Mesorhizobium genus and its importance in bacterial metabolism.
Asunto(s)
Aminopeptidasas/química , Aminopeptidasas/farmacología , Biopelículas/efectos de los fármacos , Mesorhizobium/metabolismo , Secuencia de Aminoácidos , Aminopeptidasas/genética , Aminopeptidasas/metabolismo , Activación Enzimática , Evolución Molecular , Genoma Bacteriano , Genómica/métodos , Multimerización de Proteína , Relación Estructura-Actividad Cuantitativa , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Análisis de Secuencia de ADN , SolucionesRESUMEN
Plants on contaminated mining soils often show a reduced growth due to nutrient depletion as well as trace elements (TEs) toxicity. Since those conditions threat plant's survival, plant growth-promoting rhizobacteria (PGPRs), such as rhizobia, might be of crucial importance for plant colonization on TE-contaminated soils. Native rhizobia from mining soils are promising candidates for bioaugmented phytoremediation of those soils as they are adapted to the specific conditions. In this work, rhizobia from Zn- and Cd-contaminated mining soils were in vitro screened for their PGP features [organic acids, indole-3-acetic acid (IAA), and siderophore (SID) production; 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity; and Ca3(PO4)2 solubilization] and Zn and Cd tolerance. In addition, some type and reference rhizobia strains were included in the study as well. The in vitro screening indicated that rhizobia and other native genera have great potential for phytoremediation purposes, by exerting, besides biological N2 fixation, other plant growth-promoting traits. Leucaena leucocephala-Mesorhizobium sp. (UFLA 01-765) showed multielement tolerance and an efficient symbiosis on contaminated soil, decreasing the activities of antioxidative enzymes in shoots. This symbiosis is a promising combination for phytostabilization.