Naturalproductshavebeenandcontinuetobearichsourceofleadcompoundsforthetreatmentandstudyof humandiseases.Manyofthesecompoundshaveunusualcyclicskeletonsonwhichtheirbiologicalproperties depend and often require equally unusual enzyme-catalyzed reactions for their construction. By mapping the biosynthetic pathways of these natural products and elucidating the chemical mechanisms of the reactions therein,weaimtoenrichtherepertoireoftoolsavailabletonaturalproductchemistsandsyntheticbiologistsin their efforts to develop and engineer new technologies and pharmaceuticals for the benefit of human health. However, in order to fully realize the potential of natural product biosynthesis, the pathways must be characterized, and the underlying chemistry thoroughly understood. In this spirit, we have identified three principal systems for study in the next funding period. Thus, the first specific aim is to explore the unprecedented biosynthetic pathway of ladderane lipids. The cis-fused cyclobutane ring systems of the ladderaneshavelongbeenofinteresttoscientistsgiventheirimportanceinanammoxbacteria,theirimpacton the global nitrogen cycle and their potential as biofuels. However, their biosynthesis remains enigmatic as the necessary enzyme transformations are essentially unknown and may very well involve a number of radical- mediated reactions catalyzed by radical SAM enzymes. The second specific aim seeks to understand the biological origin of two unique peptidyl nucleoside antibiotics (PNAs). Polyketide and carbohydrate biosynthesis have traditionally been considered two separate paradigms in secondary metabolism. However, recent biosynthetic investigations of amipurimycin and miharamycins have suggested that the high-carbon sugar cores of these PNAs are likely biosynthesized as polyketides.
We aim to rigorously test this hypothesis by reconstituting the biosynthetic pathways in vitro. We will determine the origin of the sugar cores in these compounds and establish the sequence and nature of the reactions involved in their construction. The third specific aim is to elucidate the pathway and reactions that are responsible for pyrazole ring formation in formycin A and pyrazofurin. The pyrazole moieties in these C-nucleoside antibiotics are notable for their N?N linkage that may require formation of an organohydrazine intermediate. However, the biological transformations underlying N?N bond formation and cyclization are presently almost entirely speculative. Thorough investigation of thesehypotheses will require the collectiveapplication of our expertise in molecular biology, chemical synthesis and enzymology to establish the biosynthetic pathways and enzymatic mechanismsofcatalysis.Thesesystemshavebeenselectedonthebasisoftheirnovelty,implicationsforthe field of mechanistic enzymology, and potential utility in biomedical research at the basic and translational levels. We believe this work will continue to address standing questions in biological chemistry and open new avenuesofdiscoveryinsecondarymetabolismandpharmaceuticalresearch.

Public Health Relevance

The proposal seeks to understand the natural formation of a number of unusual ring-containing compounds that are also notable for their established relevance with respect to antimicrobial development (amipurimycin, miharamycins, formycin A, pyrazofurin), anticancer properties (pyrazofurin) and biotechnology such as biofuels and the bioremediation of nitrogen-contaminated wastewater (ladderanes). Given their unusual chemical structures, the biosynthesis of these compounds cannot be understood in terms of currently recognized paradigms of metabolism and enzyme catalysis. This gap in understanding will be addressed by the proposal such that these and related compounds may be further developed for biomedical and biotechnological purposes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM040541-31
Application #
10050115
Study Section
Synthetic and Biological Chemistry B Study Section (SBCB)
Program Officer
Bond, Michelle Rueffer
Project Start
1989-04-01
Project End
2024-06-30
Budget Start
2020-07-01
Budget End
2021-06-30
Support Year
31
Fiscal Year
2020
Total Cost
Indirect Cost
Name
University of Texas Austin
Department
Type
Schools of Pharmacy
DUNS #
170230239
City
Austin
State
TX
Country
United States
Zip Code
78759
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