Quantifying the Effects of Frequency and Amplitude of Periodic Oxygen-Related Stress on Recombinant Protein Production in Pichia pastoris

Pichia pastoris is an attractive candidate platform for recombinant protein production. Dissolved oxygen is one of the most important factors in the cultivation of P. pastoris. However, the effect of oxygen on triggering productivity led to ambivalent results. In our earlier work, a two-compartment system, consisting of a single reactor coupled with a plug flow reactor (PFR), has been proposed as a tool to improve protein quantity and quality. The goal of this work was to investigate the effects of frequency (the residence time of broth in the PFR) and amplitude (the dissolved oxygen level in the reactor) of the stress on productivity, titer and physiology. A recombinant P. pastoris strain, which expressed horseradish peroxidase, was used as the model system. Thirteen experiments were performed. Multivariate data analysis was done and the results showed that the residence time did not influence titer, productivity and physiology over the range of residence time studied while dissolved oxygen influenced titer and specific productivity in a quadratic function. In other words, an intermediate level of dissolved oxygen (25%) showed the highest specific productivity and titter, irrespective of the residence time in the PFR. In turn, the variation of the residence time and dissolved oxygen did not influence growth physiology, as quantified in biomass and carbon dioxide yields

Experimental study of microbial enhanced oil recovery in oil-wet fractured porous media

Without Enhanced Oil Recovery (EOR) operations, the final recovery factor of most hydrocarbon reservoirs would be limited. However, EOR can be an expensive task, especially for methods involving gas injection. On the other hand, aqueous injection in fractured reservoirs with small oil-wet or mixed-wet matrices will not be beneficial if the rock wettability is not changed effectively. In the current research, an unpracticed fabrication method was implemented to build natively oil-wet, fractured micromodels. Then, the efficiency of microbial flooding in the micromodels, as a low-cost EOR method, is investigated using a new-found bacteria, Bacillus persicus. Bacillus persicus improves the sweep efficiency via reduction of water/oil IFT and oil viscosity, in-situ gas production, and wettability alteration mechanisms. In our experiments, the microbial flooding technique extracted 65% of matrix oil, while no oil was produced from the matrix system by water or surfactant flooding