(161a) Bacterial Expression of Moringa Oleifera Seed Proteins for Virus Filtration

The purpose of the work presented in this poster is to recombinantly produce Moringa oleifera proteins to understand their interactions with viral capsids in the context of water filtration. 785 million people around the world lack access to safe water (1). The leading cause of acute diarrhea in children are enteric viruses, which include rotavirus and norovirus (2). Recently, we showed that functionalizing a sand filter with crude water extract from Moringa seed can increase the virus removal efficiency by 7 orders of magnitude (3). A chitin binding protein, MOCBP, is thought to be responsible for the virus removal. An antimicrobial protein, MOCP, is also thought to affect the interactions. MOCP has previously been produced using B. subtilis to gain higher titers (4). We replicate that work for both MOCP and MOCBP. We also have also utilized a self-cleaving intein method to create a tag free version of each protein to study (5). Experiments were conducted using MS2 bacteriophage, a common surrogate for human enteric viruses. Each type of purified protein was tested independently and in combination to understand the individual and synergistic effect with virus capsid. Better understanding of the protein-virus interactions can help us design better columns and hopefully extend this research to other plant antimicrobial proteins.

References:

1. UNICEF, WHO (2019) Progress on household drinking water, sanitation and hygiene 2000-2017: Special focus on inequalities Available at: https://washdata.org/reports.
2. WHO (2008) Weekly epidemiological record Available at: http://www.who.int/vaccines-documents/DocsPDF02/www698.pdf. [Accessed April 30, 2020].
3. Samineni L, et al. (2019) 7 Log Virus Removal in a Simple Functionalized Sand Filter. Environ Sci Technol:12706–12714.
4. Arnett C, Lange J, Boyd A, Page M, Cropek D (2019) Expression and secretion of active Moringa oleifera coagulant protein in Bacillus subtilis. Appl Microbiol Biotechnol 103:9411–9422.
5. Wu W-Y, Mee C, Califano F, Banki R, Wood DW (2006) Recombinant protein purification by self-cleaving aggregation tag. Nat Protoc 1(5):2257–62.