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Date of Award

Summer 2021

Rights

Access restricted for 2 years to EWU users with an active EWU NetID

Date Available to Non-EWU Users

2023-08-13

Document Type

Thesis: EWU Only

Degree Name

Master of Science (MS) in Biology

Department

Biology

Abstract

Chronic inflammation stems from the inability of the body to resolve acute inflammation. Chronic inflammation is a growing health concern, as nearly 60% of Americans reported coping with at least one chronic condition (as of 2014). On a world scale, three fifths of deaths result from a chronic inflammation disease. Treatments for chronic inflammation include lifestyle changes (diet and exercise) and a variety of conventional drugs. Beyond these treatments, supplementing the body with systemic enzymes may attenuate inflammation. One promising enzyme is serratiopeptidase. Serratiopeptidase is a serine protease expressed by Serratia marcescens which has demonstrated anti-inflammatory, anti-edemic, and analgesic activity in laboratory studies and pre-clinical trials. Clinical trial data is less convincing, likely because the enzyme delivered is not bioavailable to the body after it passes through the stomach. To circumvent this issue, we proposed developing a plasmid vector that, when transformed into the probiotic Lactococcus lactis, would allow for bioactive serratiopeptidase production within the gut; thus, increasing bioavailability. The vector designed was delivered from the synthesis company in two fragments (they failed to assemble to full vector). After successfully, assembling the vector, the selection system in the commercial L. lactis strain we chose did not work as advertised in our hands. We introduced antibiotic resistance into the vector to facilitate the selection process after transformation. Unfortunately, the vector with antibiotic failed to transform L. lactis indicating some unknown problem with the vector / host system.

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