Xanthan gum production in Xanthomonas campestris is increased by favoring the biosynthesis of its monomers.

Current efforts to improve xanthan gum (XG) production by Xanthomonas have focused on the growth medium, operating parameters, and downstream steps. However, a key aspect is the development of optimal strains. The present work aimed to investigate the formation of XG monomers, using kinetic and stoichiometric models to identify possible bottlenecks, and to engineer a recombinant strain to overcome such limitations.

Glycerol as substrate and NADP-dependent glyceraldehyde-3-phosphate dehydrogenase enable higher production of 3-hydroxypropionic acid through the β-alanine pathway in E. coli.

Microbial engineering is a promising way to produce3-HP using biorenewable substrates such as glycerol. However, theglycerol pathway to obtain 3-HPrequires vitamin B-12, which hinders its economic viability. The present work showed that 3-HP can be efficiently produced from glycerol through the β-alanine pathway.

Immunogenicity and Protection by DnaK and SpaA Recombinant Proteins Against in a Murine Model.

Background/aims: Swine erysipelas is a disease caused by , a Gram-positive bacillus, which has great economic importance because it leads to the loss of the swine herd. To control this disease, animals are immunized with a cellular vaccine of killed or attenuated , but even with herd vaccination, cases of swine erysipelas outbreaks have been reported in the United States, China and Japan, leading to the search for other antigenic components of the bacteria that may promote greater protection against . The surface protein SpaA from has been shown to be a candidate to constitute a subunit vaccine, since it has already been reported to induce a host immune response against the bacterium.

Improving 3-hydroxypropionic acid production in E. coli by in silico prediction of new metabolic targets.

3-Hydroxypropionic acid (3-HP) production from renewable feedstocks is of great interest in efforts to develop greener processes for obtaining this chemical platform. Here we report an engineered E. coli strain for 3-HP production through the β-alanine pathway.

Antitumor Effect of IL-2 and TRAIL Proteins Expressed by Recombinant Salmonella in Murine Bladder Cancer Cells.

Background/aims: Cancer is the second most deadly disease in the world. The bladder cancer is one of the most aggressive types and shows a continuous increase in the number of cases. The use of bacteria as live vectors to deliver molecules directly to the tumor is a promising tool and has been used as an adjuvant treatment against several types of cancer.

Metabolic engineering of E. coli for pyocyanin production.

Pyocyanin is a secondary metabolite from Pseudomonas aeruginosa that belongs to the class of phenazines, which are aromatic nitrogenous compounds with numerous biological functions. Besides its antifungal and antimicrobial activities, pyocyanin is a remarkable redox-active molecule with potential applications ranging from the pharma industry to the development of microbial fuel cells. Nevertheless, pyocyanin production has been restricted to P.

Comparing the electrochemical degradation of the fluoroquinolone antibiotics norfloxacin and ciprofloxacin using distinct electrolytes and a BDD anode: evolution of main oxidation byproducts and toxicity.

The effects of the supporting electrolytes (SEs) NaSO, NaCl, NaCO, NaNO, and NaPO on the anodic oxidation of norfloxacin (NOR) and ciprofloxacin (CIPRO), assessed by the respective degradation kinetics and byproducts and electrolyzed solution antimicrobial activity, are compared. Galvanostatic anodic oxidations were performed in a filter-press flow cell fitted with a boron-doped diamond anode. Removal rates higher than the theoretical one for a process purely controlled by mass transfer were found for all SEs, indicative of contribution by indirect oxidation processes.

Electrochemical degradation of the antibiotic ciprofloxacin in a flow reactor using distinct BDD anodes: Reaction kinetics, identification and toxicity of the degradation products.

The performances of distinct BDD anodes (boron doping of 100, 500 and 2500 ppm, with sp/sp carbon ratios of 215, 325, and 284, respectively) in the electrochemical degradation of ciprofloxacin - CIP (0.5 L of 50 mg L in 0.10 M NaSO, at 25 °C) were comparatively assessed using a recirculating flow system with a filter-press reactor.

Antibiotic Korormicin A Kills Bacteria by Producing Reactive Oxygen Species.

Korormicin is an antibiotic produced by some pseudoalteromonads which selectively kills Gram-negative bacteria that express the Na-pumping NADH:quinone oxidoreductase (Na-NQR.) We show that although korormicin is an inhibitor of Na-NQR, the antibiotic action is not a direct result of inhibiting enzyme activity. Instead, perturbation of electron transfer inside the enzyme promotes a reaction between O and one or more redox cofactors in the enzyme (likely the flavin adenine dinucleotide [FAD] and 2Fe-2S center), leading to the production of reactive oxygen species (ROS).

Mapping Salmonella typhimurium pathways using C metabolic flux analysis.

In the last years, Salmonella has been extensively studied not only due to its importance as a pathogen, but also as a host to produce pharmaceutical compounds. However, the full exploitation of Salmonella as a platform for bioproduct delivery has been hampered by the lack of information about its metabolism. Genome-scale metabolic models can be valuable tools to delineate metabolic engineering strategies as long as they closely represent the actual metabolism of the target organism.

Optimization of the electrochemical degradation process of the antibiotic ciprofloxacin using a double-sided β-PbO anode in a flow reactor: kinetics, identification of oxidation intermediates and toxicity evaluation.

The electrochemical degradation of ciprofloxacin-CIP (50 mg L in 0.10 mol L NaSO) was investigated using a double-sided Ti-Pt/β-PbO anode in a filter-press flow reactor, with identification of oxidation intermediates and follow-up of antimicrobial activity against Escherichia coli. The effect of solution pH, flow rate, current density, and temperature on the CIP removal rate was evaluated.

The effect of the supporting electrolyte on the electrooxidation of enrofloxacin using a flow cell with a BDD anode: Kinetics and follow-up of oxidation intermediates and antimicrobial activity.

The role of the supporting electrolyte - SE (NaSO; NaCl; NaCO; NaNO; NaPO - 0.1 M ionic strength) in the galvanostatic (10 mA cm) electrochemical degradation of the fluoroquinolone antibiotic enrofloxacin (ENRO; 100 mg L) using a filter-press flow cell with a boron-doped diamond anode was investigated (flow rate, solution volume, and temperature were kept fixed at 420 L h, 1.0 L, and 25 °C, respectively).

One-pot strategy for on-site enzyme production, biomass hydrolysis, and ethanol production using the whole solid-state fermentation medium of mixed filamentous fungi.

The efficient use of renewable lignocellulosic feedstocks to obtain biofuels and other bioproducts is a key requirement for a sustainable biobased economy. This requires novel and effective strategies to reduce the cost contribution of the cellulolytic enzymatic cocktails needed to convert the carbohydrates into simple sugars, in order to make large-scale commercial processes economically competitive. Here, we propose the use of the whole solid-state fermentation (SSF) medium of mixed filamentous fungi as an integrated one-pot strategy for on-site enzyme production, biomass hydrolysis, and ethanol production.

Electrochemical mineralization of cephalexin using a conductive diamond anode: A mechanistic and toxicity investigation.

The contamination of surface and ground water by antibiotics is of significant importance due to their potential chronic toxic effects to the aquatic and human lives. Thus, in this work, the electrochemical oxidation of cephalexin (CEX) was carried out in a one compartment filter-press flow cell using a boron-doped diamond (BDD) electrode as anode. During the electrolysis, the investigated variables were: supporting electrolyte (NaSO, NaCl, NaNO, and NaCO) at constant ionic strength (0.

Salmonella typhimurium and Escherichia coli dissimilarity: Closely related bacteria with distinct metabolic profiles.

Live attenuated strains of Salmonella typhimurium have been extensively investigated as vaccines for a number of infectious diseases. However, there is still little information available concerning aspects of their metabolism. S.

High-throughput strategies for penicillin G acylase production in rE. coli fed-batch cultivations.

Background: Penicillin G acylase (PGA) is used industrially to catalyze the hydrolysis of penicillin G to obtain 6-aminopenicillanic acid. In Escherichia coli, the most-studied microorganism for PGA production, this enzyme accumulates in the periplasmic cell space, and temperature plays an important role in the correct synthesis of its subunits.

Results: This work investigates the influence of medium composition, cultivation strategy, and temperature on PGA production by recombinant E.

Live bacterial vaccine vectors: an overview.

Genetically attenuated microorganisms, pathogens, and some commensal bacteria can be engineered to deliver recombinant heterologous antigens to stimulate the host immune system, while still offering good levels of safety. A key feature of these live vectors is their capacity to stimulate mucosal as well as humoral and/or cellular systemic immunity. This enables the use of different forms of vaccination to prevent pathogen colonization of mucosal tissues, the front door for many infectious agents.

Non-conventional induction strategies for production of subunit swine erysipelas vaccine antigen in rE. coli fed-batch cultures.

In spite of the large number of reports on fed-batch cultivation of E. coli, alternative cultivation/induction strategies remain to be more deeply exploited. Among these strategies, it could be mentioned the use of complex media with combination of different carbon sources, novel induction procedures and feed flow rate control matching the actual cell growth rate.

Enhanced production of recombinant thermo-stable lipase in Escherichia coli at high induction temperature.

Thermostable microbial lipases are potential candidates for industrial applications such as specialty organic syntheses as well as hydrolysis of fats and oils. In this work, basic biochemical engineering tools were applied to enhance the production of BTL2 lipase cloned in Escherichia coli BL321 under control of the strong temperature-inducible λP(L) promoter. Initially, surface response analysis was used to assess the influence of growth and induction temperatures on enzyme production, in flask experiments.

Cloning, auto-induction expression, and purification of rSpaA swine erysipelas antigen.

This work reports the cloning, expression, and purification of a 42-kDa fragment of the SpaA protein from Erysipelothrix rhusiopathiae, the main antigenic candidate for a subunit vaccine against swine erysipelas. The use of an auto-induction protocol to improve heterologous protein expression in recombinant Escherichia coli cultures was also investigated. The cellular growth pattern and metabolite formation were evaluated under different induction conditions.

Intensification of high cell-density cultivations of rE. coli for production of S. pneumoniae antigenic surface protein, PspA3, using model-based adaptive control.

This work proposes an innovative methodology to control high density fed-batch cultures of E. coli, based on measurements of the concentration of dissolved oxygen and on estimations of the cellular specific growth rate (µ), of the yield of biomass/limiting substrate (Y (xs)) and of the maintenance coefficient (m). The underlying idea is to allow cells to grow according to their metabolic capacity, without the constraints inherent to pre-set growth rates.

Robust artificial intelligence tool for automatic start-up of the supplementary medium feeding in recombinant E. coli cultivations.

One of the most important events in fed-batch fermentations is the definition of the moment to start the feeding. This paper presents a methodology for a rational selection of the architecture of an artificial intelligence (AI) system, based on a neural network committee (NNC), which identifies the end of the batch phase. The AI system was successfully used during high cell density cultivations of recombinant Escherichia coli.

Bioreactor aeration conditions modulate growth and antigen expression during Erysipelothrix rhusiopathiae cultivation.

Erysipelothrix rhusiopathiae, the causative agent of swine erysipelas, was cultivated in a 5-L stirred and aerated bioreactor under different dissolved oxygen tensions (0%, 5%, and 30% of saturation) for evaluation of the influence of oxygen on cell growth as well as on the production of the main antigenic component of the vaccine against erysipelas, a 64-69 kDa protein (SpaA). The microorganism presented different growth profiles for different aeration conditions. However, at the end of the batch cultivations, similar cell concentrations were obtained under the studied conditions.