The two faces of pyocyanin - why and how to steer its production?

The ambiguous nature of pyocyanin was noted quite early after its discovery. This substance is a recognized Pseudomonas aeruginosa virulence factor that causes problems in cystic fibrosis, wound healing, and microbiologically induced corrosion. However, it can also be a potent chemical with potential use in a wide variety of technologies and applications, e.g. green energy production in microbial fuel cells, biocontrol in agriculture, therapy in medicine, or environmental protection. In this mini-review, we shortly describe the properties of pyocyanin, its role in the physiology of Pseudomonas and show the ever-growing interest in it. We also summarize the possible ways of modulating pyocyanin production. We underline different approaches of the researchers that aim either at lowering or increasing pyocyanin production by using different culturing methods, chemical additives, physical factors (e.g. electromagnetic field), or genetic engineering techniques. The review aims to present the ambiguous character of pyocyanin, underline its potential, and signalize the possible further research directions.

The use of the electromagnetic field in microbial process bioengineering.

An electromagnetic field (EMF) has been shown to have various stimulatory or inhibitory effects on microorganisms. Over the years, growing interest in this topic led to numerous discoveries suggesting the potential applicability of EMF in biotechnological processes. Among these observations are stimulative effects of this physical influence resulting in intensified biomass production, modification of metabolic activity, or pigments secretion. In this review, we present the current state of the art and underline the main findings of the application of EMF in bioprocessing and their practical meaning in process engineering using examples selected from studies on bacteria, archaea, microscopic fungi and yeasts, viruses, and microalgae. All biological data are presented concerning the classification of EMF. Furthermore, we aimed to highlight missing parts of contemporary knowledge and indicate weak spots in the approaches found in the literature.

Application of Magnetically Assisted Reactors for Modulation of Growth and Pyocyanin Production by Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a producer of desired secondary metabolites, including pyocyanin. Potential uses of this pigment urge a search for improved production methods. Recent trends in bioprocessing show the potential of the use of electromagnetic fields (EMFs) to influence the growth of microorganisms and even modulate the concentration of bioproducts. Here, we aimed at assessing the influence of rotating magnetic field (RMF) and static magnetic field (SMF) on pyocyanin production, growth rate, and respiration of P. aeruginosa. Moreover, exposure time to EMFs (2, 6, and 12 h) and culture volume (10 and 50 ml) were initially assessed. P. aeruginosa was cultivated in magnetically assisted reactors with 5 and 50 Hz RMF (magnetic induction of 24.32 and 42.64 mT, respectively) and SMF (-17.37 mT). Growth kinetics was assessed with Gompertz equation. The viability was tested using resazurin assay, whereas pyocyanin production by chloroform-HCl methodology. The growth of P. aeruginosa was slightly stimulated by exposure to a RMF with 50 Hz (108% related to the control) and significantly by SMF (132% related to the control), while RMF 5 Hz exposure prolonged the time of inflection (in comparison to RMF 50 Hz and SMF). The 6-h exposure to EMFs resulted in the highest pyocyanin production in comparison to the control, indicating a relationship between exposure time and product concentration. Moreover, cultures led in smaller volumes produced more pyocyanin. Our findings show that the use of different EMF types, frequency, and exposition time and volume could be used interchangeably to obtain different bioprocess aims.

The Influence of Hydrodynamic Conditions in a Laboratory-Scale Bioreactor on Pseudomonas aeruginosa Metabolite Production.

Hydrodynamic conditions are critical in bioprocessing because they influence oxygen availability for cultured cells. Processes in typical laboratory bioreactors need optimization of these conditions using mixing and aeration control to obtain high production of the desired bioproduct. It could be done by experiments supported by computational fluid dynamics (CFD) modeling. In this work, we characterized parameters such as mixing time, power consumption and mass transfer in a 2 L bioreactor. Based on the obtained results, we chose a set of nine process parameters to test the hydrodynamic impact on a selected bioprocess (mixing in the range of 0-160 rpm and aeration in the range of 0-250 ccm). Therefore, we conducted experiments with P. aeruginosa culture and assessed how various hydrodynamic conditions influenced biomass, pyocyanin and rhamnolipid production. We found that a relatively high mass transfer of oxygen (kLa = 0.0013 s-1) connected with intensive mixing (160 rpm) leads to the highest output of pyocyanin production. In contrast, rhamnolipid production reached maximal efficiency under moderate oxygen mass transfer (kLa = 0.0005 s-1) and less intense mixing (in the range of 0-60 rpm). The results indicate that manipulating hydrodynamics inside the bioreactor allows control of the process and may lead to a change in the metabolites produced by bacterial cells.

Development of an Autochthonous Microbial Consortium for Enhanced Bioremediation of PAH-Contaminated Soil.

The main objectives of this study were to isolate bacteria from soil chronically contaminated with polycyclic aromatic hydrocarbons (PAHs), develop an autochthonous microbial consortium, and evaluate its ability to degrade PAHs in their native contaminated soil. Strains with the best bioremediation potential were selected during the multi-stage isolation process. Moreover, to choose bacteria with the highest bioremediation potential, the presence of PAH-degrading genes (pahE) was confirmed and the following tests were performed: tolerance to heavy metals, antagonistic behavior, phytotoxicity, and antimicrobial susceptibility. In vitro degradation of hydrocarbons led to the reduction of the total PAH content by 93.5% after the first day of incubation and by 99.22% after the eighth day. Bioremediation experiment conducted in situ in the contaminated area resulted in the average reduction of the total PAH concentration by 33.3% after 5 months and by over 72% after 13 months, compared to the concentration recorded before the intervention. Therefore, this study implicates that the development of an autochthonous microbial consortium isolated from long-term PAH-contaminated soil has the potential to enhance the bioremediation process.