(620bx) Engineering Pseudomonas Fluorescens for Efficient Bioseparation
P. fluorescens is slowly gaining popularity as a protein expression platform principally due to its low endotoxin potential and relative ease of cultivation. However, little is known about the proteome of this organism as it relates to downstream processing, as well as how targeted alterations can be used to mitigate the expression of nuisance proteins. To this end, this poster describes our effort to enhance the bioprocessing potential of P. fluorescens through rational host cell engineering. Host cell proteins were identified within the proteome that are retained strongly by ion exchange chromatography media, the data on which was used to guide recombineering efforts. Lack of an identified CRISPR system prompted our group to import elements of this mechanism for gene targeting in pseudomonas, used to delete or tune repression of proteins which were demonstrated to reduce column capacity and complicate gradient elution.