Date of Award

2023-08-01

Degree Name

Master of Science

Department

Chemistry

Advisor(s)

Chu-Young Kim

Abstract

Bioactive natural products often possess a complex structure and have a large size which make their chemical synthesis highly challenging. To overcome this challenge, we are developing a biosynthetic method for production of natural products and their analogs. This technology is expected to decrease the synthesis time from years to weeks and reduce the cost of drug production by multiple orders of magnitude. We are investigating the biosynthesis pathway of lasalocid A, an anticancer antibiotic, to develop this technology. Specifically, we aim to characterize the enzymes structurally and biochemically in this pathway, thus paving the way for their rational engineering. We have expressed and purified the recombinant Lsd13 and Lsd14 polyketide synthases, two key enzymes from the lasalocid A biosynthesis pathway. We have also obtained potential crystals of the Lsd13 protein and potential crystals of the partial Lsd13-Lsd14 complex. We will use these crystals to perform X-ray diffraction experiments in the future and ultimately determine their atomic structure. We have also characterized the binding interaction between Lsd13 and Lsd14 using surface plasmon resonance spectroscopy. Our results suggest that Lsd13 and Lsd14 are dimeric proteins and that they form a stable complex to allow transfer of the intermediate polyketide chain product from Lsd13 to Lsd14.

Language

en

Provenance

Recieved from ProQuest

File Size

54 p.

File Format

application/pdf

Rights Holder

Dayan Viera

Included in

Biochemistry Commons

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