Effect of Indole-3-Acetic Acid on Tomato Plant Growth.

Plant growth-promoting bacteria have several abilities to promote plant growth and development. One of these skills is the synthesis of indole-3-acetic acid (IAA), which mainly promotes root and shoot development. The bacteria Bacillus subtilis and Azospirillum brasilense have been widely used in agriculture with this function. However, little is known about whether the joint inoculation of these bacteria can reduce plant development by the excess of IAA produced as a result of the joint inoculation. The objective of the present study was to verify the effect of IAA on the inoculation of B. subtilis and A. brasilense in three tomato genotypes. The Micro-Tom genotype without mutation for IAA synthesis, Entire, has high sensitivity to IAA, and the diageotropic genotype (dgt) has low sensitivity to IAA. The results show that the plant parameter most sensitive to microbial inoculation is the number of roots. No treatment increased the shoot dry mass parameters for the Micro-Tom genotype and dgt, root dry mass for the Micro-Tom genotype, plant height for the Micro-Tom and Entire genotypes, root area and root volume for the genotype dgt. The Azm treatment reduced plant height compared to the control in the dgt, the BS + Azw and BS + Azm treatments in the Micro-Tom genotype and the Azw + Azm treatment in the dgt genotype reduced the plant diameter compared to the control. BS and BS + Azw reduced the number of roots in the Micro-Tom. The results strongly support that the mixture of B. subtilis and A. brasilense can reduce some parameters of plant development; however, this effect is possibly an interference in the mode of action of growth promotion of each isolate and is not related to an excess of IAA produced by the bacteria.

Bioremediation of heavy metal-polluted environments by non-living cells from rhizobial isolates.

Rhizosphere bacteria, for example, rhizobia, can play several roles, and one of the most important, the protection of plant roots against toxic conditions and other environmental stresses. In this work, the action of Cu2+ and Cr6+ on cell growth and EPS production of four strains of rhizobia, Rhizobium tropici (LBMP-C01), Ensifer sp. (LBMP-C02 and LBMP-C03), and Rhizobium sp. LBMP-C04, were tested. The results confirmed the strong effect of Cu2+ and Cr6+ on bacterial exopolysaccharides (EPS) synthesis, and how cells can adsorb these metals, which may be a key factor in the interactions between rhizosphere bacteria and host plants in heavy metal-contaminated soils. Here, we emphasize the importance of proving the potential of treating bacterial cells and their extracellular EPS to promote the bio-detoxification of terrestrial and aquatic systems contaminated by heavy metals in a highly sustainable, economic, and ecological way.

Preparation of Aspergillus niger 426 naringinases for debittering citrus juice utilization of agro-industrial residues.

Naringin, considered as the main bitter component of grapefruit, requires the use of enzymes to reduce the level of this substance during juice processing. For this reason, it has been the focus of many studies. In this study, to increase the production of naringinase by Aspergillus niger cultivated in solid-state fermentation (SSF), a three-component simplex-centric mixing design along with a response surface methodology (RSM) was applied to generate statistical models and analyze the dataset. First, grapefruit peel, rice bran, and wheat bran were used for substrate selection for naringinase production and, finally, selected the best of the three inducers or their mixtures to remove the bitterness of grapefruit juice. Cultivation with 2.3 g of grapefruit peel, 2.5 g of rice bran, and 5.2 g of wheat bran and medium supplementation with a mixture of naringin, rutin, and hesperidin in the concentration of 2, 5, 4.5, and 3.0 g/L, respectively, resulted in a maximum activity of 28 U/mL. The results indicate that the sequencing procedure, which allowed the definition of an optimal mixture of components, is a new way for microorganisms to have a high naringinase yield, in particular by SSF, since our data showed a 96% increase in the production of naringinase. This dataset can help other researchers apply a mixing design to increase enzyme production.

The potential of extracellular biopolymer production by Mesorhizobium sp. from monosaccharide constituents of lignocellulosic biomass.

OBJECTIVE: The effects of monosaccharide constituents of lignocellulosic materials on exopolysaccharide (EPS) production by Mesorhizobium sp. Semia 816 were studied. RESULTS: According to the results, by using sugars commonly found in lignocellulosic biomass as carbon sources (glucose, arabinose and xylose), no significant differences were observed in the production of EPS, reaching 3.39 g/L, 3.33 g/L and 3.27 g/L, respectively. Differences were observed in monosaccharide composition, mainly in relation to rhamnose and glucuronic acid contents (1.8 times higher when arabinose was compared with xylose). However, the biopolymers showed no differences in relation to rheological properties, with EPS aqueous-based suspensions (1.0% w/v) presenting pseudoplastic behavior, and a slight difference in degradation temperatures. Using soybean hulls hydrolysate as carbon source, slightly higher values were obtained (3.93 g/L). CONCLUSION: The results indicate the potential of the use of lignocellulosic hydrolysates containing these sugars as a source of carbon in the cultivation of Mesorhizobium sp. Semia 816 for the production of EPS with potential industrial applications.

Bagasse minority pathway expression: Real time study of GH2 ß-mannosidases from bacteroidetes.

After being isolated from a sugarcane pile, the bacterium Chitinophaga sp. CB10 demonstrated to be a rich source of carbohydrases, with 350 predicted CAZyme domains. CB10 was able to grow on carbohydrates of different structural complexities: glucose, carboxymethylcellulose, corn starch, galactomannan, Aloe vera gum and sugarcane bagasse. The sugarcane bagasse is a rich source of complex polymers, and the diversity of metabolites released by its enzymatic hydrolysis has an important role for green chemistry, including minority pathways such as the degradation of mannan conjugates. In this sense, CB10 demonstrated considerable levels of gene expression for mannanases, and was stable for a period of 96-144 hours in the presence of sugarcane bagasse as sole carbon source. The bacterium showed respectively 4.8x and 5.6x expression levels for two genes predicted for GH2 ß-mannosidase: one located within a gene cluster identified as "polysaccharide utilization loci" (PUL), and another a classic ß-mannosidase. These enzymes shared less than 45% of identity with enzymes characterized from the genus Chitinophaga belonging to the phylum Bacteroidetes. The degree of novelty-as demonstrated by the low identity with previously characterized enzymes; the remarkable capability to grow in different substrates; mannanase activity, evidenced by the release of residual oligosaccharides in the cultivation with galactomannan (HPLC-RID, 12.3 mMol); associated to the ability of mannanases expression in a low concentration of inductor conditions (sugarcane bagasse, 0.2%) indicate the high potential for the application of CB10 as a source of enzymes in the production of oligosaccharides from biomass. This capacity might prove to be very valuable for the biorefinery process of pre-biotic precursors and other functional oligosaccharides focused on the food and pharmaceutical industries.

Exploring the Potential of Two Bacterial Consortia to Degrade Cellulosic Biomass for Biotechnological Applications.

This paper outlines two cellulolytic bacterial consortia named SCS and SCB, isolated from soil samples of sugarcane (Saccharum officinarum) crop field, and a sugarcane bagasse deposit in an ethanol mill. Both consortia were able to grow on different carbon sources, such as sugarcane bagasse, corn husk, peanut hulls, and carboxymethylcellulose, releasing up to 11.90 µmol/mL and 15.23 µmol/mL of glucose for SCS and SCB, respectively. In addition, SCS and SCB have several strains capable of producing cellulase, amylase, lipase, and protease. Whole genome sequencing of the SCS consortium revealed that Burkholderia was the most prevalent genus, encompassing approximately 80% of the consortia. In addition, metagenome analysis allowed the identification of genes encoding enzymes related to starch and cellulose degradation, as well as enzymes related to lipases and proteases, confirming our initial findings. The results showed that SCS and SCB had the capability to degrade cellulose, and that they were an efficient source of enzyme production, which would provide a new choice for use in different biotechnological applications.

Rhizobium spp exopolysaccharides production and xanthan lyase use on its structural modification.

Enzymes can be very useful on exopolysaccharides (EPS) research, can be used at elucidation and also to modify the polysaccharides' structure in order to alter their physical properties. Thus, the reduction of the molecular mass could increase applications of these biopolymers. Therefore, the EPS production of different rhizobia and the action of xanthan lyase on its structures were evaluated. The strains produced significant amounts of EPS, and it was noticed that are heteropolysaccharides, composed galactose and glucose. Both EPS and xanthan were modified on ß-glycosidic bonds, the mannose was removed of xanthan had but the EPS was affected in the CO stretching vibration, where the glucuronic acid removed from of your structure. The ester/carboxylic acid portions affected functional groups of the acetate/succinate, methyl carbons of the O-acetyl and pyruvate methyl groups in addition to affect the carbons the main pyranoid. The Resistance to temperature increase of the EPS was observed, made possible by the activity of the lyase. EPS has the ability to form stable gels at higher temperatures and anionic feature can be used on solubilization and controlled release of substances. Modified EPS knowledge will presently facilitate future investigations relating the structure of the rhizobia polysaccharide against rheological properties.

Characterization of new exopolysaccharide production by Rhizobium tropici during growth on hydrocarbon substrate.

Exopolysaccharide (EPS) are produced by a diverse of rhizobia species and has been demonstrated to be a bioemulsifier with potential applications in the degradation of hydrocarbons. In the present study, attempts were made to obtain the new exopolysaccharide production by Rhizobium tropici (SEMIA 4080 and MUTZC3) strains during growth on hydrocarbon substrate. Under the different cultivation conditions, the high molecular weight exopolysaccharides from Rhizobium tropici strains cultivated for 96h mainly consisted of carbohydrates (79-85%) and a low percentage of protein. The EPSC3-D differed from the others, with only 60% of carbohydrate. However, all strains produced polymers with distinct rheology properties, such as viscosity of each EPS sample, suitable for different applications. In addition, RP-HPLC, FTIR and NMR studies revealed EPS produced by rhizobia strains were similar indicating minimal difference between EPS compositions.

Bg10: A Novel Metagenomics Alcohol-Tolerant and Glucose-Stimulated GH1 ß-Glucosidase Suitable for Lactose-Free Milk Preparation.

New ß-glucosidases with product (glucose) or ethanol tolerances are greatly desired to make industrial processes more marketable and efficient. Therefore, this report describes the in silico/vitro characterization of Bg10, a metagenomically derived homodimeric ß-glucosidase that exhibited a Vmax of 10.81 ± 0.43 µM min-1, Kcat of 175.1± 6.91 min-1, and Km of 0.49 ± 0.12 mM at a neutral pH and 37°C when pNP-ß-D-glucopyranoside was used as the substrate, and the enzyme retained greater than 80% activity within the respective pH and temperature ranges of 6.5 to 8.0 and 35 to 40°C. The enzyme was stimulated by its product, glucose; consequently, the Bg10 activity against 50 and 100 mM of glucose were increased by 36.8% and 22%, respectively, while half of the activity was retained at 350 mM. Moreover, the Bg10 was able to hydrolyse 55% (milk sample) and 100% (purified sugar) of the lactose at low (6°C) and optimum (37°C) temperatures, respectively, suggesting the possibility of further optimization of the reaction for lactose-free dairy production. In addition, the enzyme was able to fully hydrolyse 40 mM of cellobiose at one hour and was tolerant to ethanol up to concentrations of 500 mM (86% of activity), while a 1 M concentration still resulted in 41% residual activity, which could be interesting for biofuel production.

Optimized medium culture for Acidobacteria subdivision 1 strains.

Members of subdivision 1 of the phylum Acidobacteria were grown at different pH values in a new medium formulation named PSYL 5, which includes sucrose as a carbon source and other compounds (such as KH2PO4 and MgSO4.7H2O). Growth rate was nearly constant at pH 5.0 and declined at pH 3-4 and 6-7. However, it was found that effects involving good carbon/nitrogen ratios and pH on the growth of the members of Acidobacteria subdivision 1 were significant, and the strongest effect of these conditions was at pH 5.0. In addition, incubation time of 48, 72, 96 and 120 h was shorter than that described previously for members of Acidobacteria subdivision 1 on solid laboratory media.

Properties of Polyhydroxyalkanoate Granules and Bioemulsifiers from Pseudomonas sp. and Burkholderia sp. Isolates Growing on Glucose.

A Burkholderia and Pseudomonas species designated as AB4 and AS1, respectively, were isolated from soil containing decomposing straw or sugar cane bagasse collected from Brazil. This study sought to evaluate the capacities of culture media, cell-free medium, and crude lysate preparations (containing PHB inclusion bodies) from bacterial cell cultures to stabilize emulsions with several hydrophobic compounds. Four conditions showed good production of bioemulsifiers (E24 ≥ 50 %), headed by substantially cell-free media from bacterial cell cultures in which bacterial isolates from Burkholderia sp. strain AB4 and Pseudomonas sp. strain AS1 were grown. Our results revealed that the both isolates (AB4 and AS1 strains) exhibited high emulsification indices (indicating usefulness in bioremediation) and good stabilities.

Chemical and rheological properties of exopolysaccharides produced by four isolates of rhizobia.

The rheological, physicochemical properties, emulsification and stability of exopolysaccharides (EPSs) from four rhizobia isolates (LBMP-C01, LBMP-C02, LBMP-C03 and LBMP-C04) were studied. The EPS yields of isolates under these experimental conditions were in the range of 1.5-6.63gL(-1). The LBMP-C04 isolate, which presented the highest EPS production (6.63gL(-1)), was isolated from Arachis pintoi and was identified as a Rhizobium sp. strain that could be explored as a possible potential source for the production of extracellular heteropolysaccharides. All polymers showed a pseudoplastic non-Newtonian fluid behavior or shear thinning property in aqueous solutions. Among the four EPS tested against hydrocarbons, EPS LBMP-C01 was found to be more effective against hexane, olive and soybean oils (89.94%, 82.75% and 81.15%, respectively). Importantly, we found that changes in pH (2-11) and salinity (0-30%) influenced the emulsification of diesel oil by the EPSs. EPSLBMP-C04 presented optimal emulsification capacity at pH 10 (E24=53%) and 30% salinity (E24=27%). These findings contribute to the understanding of the influence of the chemical composition, physical properties and biotechnology applications of rhizobial EPS solutions their bioemulsifying properties.

Production of exopolysaccharide from rhizobia with potential biotechnological and bioremediation applications.

The potential use of rhizobia under controlled fermentation conditions may result in the production of new extracellular polymeric substances (EPS) having novel and superior properties that will open up new areas of industrial applications and thus increase their demand. The production of EPS and the stability of emulsions formed with soybean oil, diesel oil and toluene using different concentrations of purified EPS derived from wild-type and mutant strains of Rhizobium tropici SEMIA 4077 was investigated. The EPS was defined as a heteropolysaccharide composed of six constituent monosaccharides that displayed higher intrinsic viscosity and pseudoplastic non-Newtonian fluid behavior in an aqueous solution. The ratio between the total EPS production and the cellular biomass was 76.70 for the 4077::Z04 mutant strain and only 8.10 for the wild-type strain. The EPS produced by the wild-type R. tropici SEMIA 4077 resulted in more stable emulsions with the tested toluene than xanthan gum, and the emulsification indexes with hydrocarbons and soybean oil were higher than 50%, indicating strong emulsion-stabilizing capacity. These results demonstrate that the EPS of R. tropici strains could be attractive for use in industrial and environmental applications, as it had higher intrinsic viscosity and good emulsification activity.

Evaluation of the biotechnological potential of Rhizobium tropici strains for exopolysaccharide production.

Rhizobium tropici, a member of the Rhizobiaceae family, has the ability to synthesize and secrete extracellular polysaccharides (EPS). Rhizobial EPS have attracted much attention from the scientific and industrial communities. Rhizobial isolates and R. tropici mutants that produced higher levels of EPS than the wild-type strain SEMIA4080 were used in the present study. The results suggested a heteropolymer structure for these EPS composed by glucose and galactose as prevailing monomer unit. All EPS samples exhibited a typical non-Newtonian and pseudoplastic fluid flow, and the aqueous solutions apparent viscosities increased in a concentration-dependent manner. These results serve as a foundation for further studies aimed at enhancing interest in the application of the MUTZC3, JAB1 and JAB6 strains with high EPS production and viscosity can be exploited for the large-scale commercial production of Rhizobial polysaccharides.