Bioengineering a non-pathogenic bacteria to produce medically-relevant biopolymers

DESCRIPTION provided by applicant Alginate is a commercially viable anionic biopolymer consisting of two monomeric subunits D mannuronate and a L guluronate Alginate is used in a variety of applications including textile manufacturing waste water remediation food production and a variety of medical applications such as wound care and drug delivery Currently commercial alginate is harvested from various genera of brown seaweeds However seaweed derived alginate has several limitations including seasonal growth inconsistencies in yield composition and quality between harvests supply side instability and the use of economically and environmentally costly processing methods These limitations impact seaweed derived alginateandapos s use for biomedical and pharmaceutical application Alginate is also produced by bacteria in the genera Azotobacter and Pseudomonas Moreover bacteria derived alginate has several advantages specifically it is relatively easy inexpensive and environmentally friendly to cultivate and harvest Additionally the bacteria can be easily manipulated to produce large amounts of alginate with specific customized properties for biomedical and pharmaceutical applications However there is little ongoing research to develop bacterial strains for commercial alginate production Progenesis Technologies LLC has patented a stable high alginate producing strain of Pseudomonas aeruginosa for use in recombinant alginate production At a pre petition consultation the United States Federal Drug Administration FDA suggested abrogating the function of the secreted virulence factors exotoxin A and pyocyanin to decrease the relative pathogenicity of VE The FDA also suggested using the currently approved bacterial strains for alginate production To date we have abrogated the function of the virulence factors exotoxin A pyocyanin phospholipase C and endotoxin In this proposal we intended to design and evaluate a non pathogenic bacterial strain for the production of the biopolymer alginate We propose to accomplish this through the completion of two Specific Aims In Aim we will evaluate the pathogenicity of our recently modified P aeruginosa strain PGN using a murine model In Aim we will determine the feasibility of producing alginate using currently approved E coli strains At the conclusion of ths project we will seek the appropriate federal approval to use our newly acquired non pathogenic bacterial strain as a platform for the production of biomedical and pharmaceutical grade alginate for use in various medical applications PUBLIC HEALTH RELEVANCE The primary goal of Progenesis Technologies LLC is to genetically engineer bacteria to produce the biopolymer alginate at a cost that is competitive with seaweed derived alginate Our technology will lead to the development of a product commonly used in a variety of biomedical and pharmaceutical applications which has enhanced performance due to our ability to alter the composition of the biopolymer and is manufactured in an environmentally friendly manner Our genetically engineered alginates can have a positive impact on public health by aiding in the care and recovery of patients that require specialized wound care topical medications and time released drug delivery