Statistical screening of medium components for recombinant production of Pseudomonas aeruginosa ATCC 9027 rhamnolipids by nonpathogenic cell factory Pseudomonas putida KT2440.

Rhamnolipids (RLs) produced by the opportunistic human pathogen Pseudomonas aeruginosa are considered as potential candidates for the next generation of surfactants. Large-scale production of RLs depends on progress in strain engineering, medium design, operating strategies, and purification procedures. In this work, the rhlAB genes extracted from a mono_RLs_producing strain of P. aeruginosa (ATCC 9027) were introduced to an appropriate safety host Pseudomonas putida KT2440. The capability of the recombinant strain was evaluated in various media. As a prerequisite for optimal medium design, a set of 32 experiments was performed in two steps for screening a number of macro-nutritional compounds. In the experiments, a two-level fractional factorial design resolution IV was followed by a two-level full factorial one. By means of this approach, it was observed that glycerol, yeast extract, and peptone have significant positive influence on recombinant RLs production while the yeast extract/peptone two-factor and glycerol/yeast extract/peptone three-factor interactions have considerable negative effects. A wide range of variation from 0 to 570 mg/l was obtained for RLs production during the screening experiments indicating the importance of medium optimization. The results point out the opportunity for possible higher yields of RLs through further screening, mixture/combined mixture designs, and high-cell-density cultivations.

Investigating wettability alteration during MEOR process, a micro/macro scale analysis.

Wettability alteration is considered to be one of the important mechanisms that lead to increased oil recovery during microbial enhanced oil recovery (MEOR) processes. Changes in wettability will greatly influence the petrophysical properties of the reservoir rocks and determine the location, flow and distribution of different fluids inside the porous media. Understanding the active mechanisms of surface wettability changes by the bacteria would help to optimize the condition for more oil recovery. As the mechanisms behind wettability alteration are still poorly understood, the objective of this study is to investigate the wettability alteration at pore scale and find the most effective mechanism of wettability changes in different cases. The experiments were performed on different substrates at fresh condition or aged in crude oil to mimic various wetting conditions. Using an Enterobacter cloacae strain, the influence of bacterial metabolites, bacterial adhesion and bacterial solution with two different carbon sources on wettability were determined for different aging periods. Contact angle measurements were used to quantify the wettability alteration of the solid surfaces. Atomic force microscopy (AFM) experiments were also utilized to combine the macroscopic measurements of wettability with the microscopic study of the surface changes. It was found that the surface wettability could vary from neutral- or oil-wet to water-wet state. Bacterial adhesion and biofilm formation seems to be the dominant mechanism of wettability alteration. The aged glass surfaces regained their initial water wetness where the bacteria could remove the polar and asphaltene compounds from them.