The CBB supports a seed grant program designed to encourage innovative and interdisciplinary research activities in the Biocatalytic Sciences. This program is supported through a combination of funding from the State of Iowa as well as revenue generated from the Center’s Microbial Fermentation and Bioprocessing Facility.
The objectives of this seed grant program are to: 1) stimulate new federal grants by supporting early-stage research efforts; 2) strengthen graduate programs through interdisciplinary studies; 3) support research teams across disciplines to provide unique approaches for solving long-standing scientific questions; and 4) create interest in the submission of federally supported center grants.
Biocatalytic Sciences is broadly defined as experimental and computational efforts, both basic and applied, that advance the fundamental understanding and general utility of processes intrinsically dependent upon biocatalytic elements, such as cells, enzymes, RNA, and DNA. Examples include: enzyme mechanisms, biochemical pathways, bioprocess monitoring, molecular biology processes, drug target development, cellular engineering, genetic engineering, metabolomics, transcriptomics, synthetic biology, green manufacturing (fuels, foods, materials), environmental remediation, medicines (vaccines, biologics, medicinal microbiome, genetic correction of diseases), and devices.
Applications are encouraged for seed projects designed to advance knowledge in the Biocatalytic Sciences or to initiate efforts to pursue a center grant. Applicants must be either current members of the CBB faculty or University of Iowa faculty interested in joining the CBB. Only faculty with either tenured or tenure-track appointments are eligible to apply. Budgets are limited to a maximum of $35,000 (direct costs only) for a one-year effort. We anticipate funding three to five awards for FY21. Previous awardees are not eligible to submit an application for two years after an award.
Proposal Deadline: Funding for seed grants in FY24 will be determined at a later date. Contact Mitch Rotman at mitchell-rotman@uiowa.edu if you have any questions.
Second year Seed Grants Awardees- August 1, 2019-July 31, 2020
Mechanoenzymatic Hydroxylation of Steroids with Cytochrome P450 CYP3A4- Leonard MacGillivray, Professor, Department of Chemistry $34,837
Collaborator: Shuvedu Das, Associate Research Scientist, Center for Biocatalysis and Bioprocessing (CBB)
Executive Summary
We propose to apply mechanochemistry to selectively hydroxylate steroids using CYP3A4 from
the cytochrome P450 family. The application of mechanochemistry to biochemical processes
is a burgeoning area, yet there has been no attention placed on chemical transformations of
biocatalytic importance. Steroids are extremely important industrially (e.g. pharmaceutical, fine
chemical). Biochemical processes that perform hydroxylations of steroids with high efficiency
and selectivity are favored to functionalize C-H bonds. The generally poor solubility of steroids
in aqueous media considerably limits conversions by enzymes. Mechanochemical approaches
to enzymatic transformations can provide efficient and clean solutions to tackle modern
problems of biocatalysis. We propose to study the application and scope of ball milling for 6β-
hydroxylation of testosterone using CYP3A4. Professor MacGillivray will make use of his
expertise in mechanochemistry. The work will be conducted with Dr. Das who has extensive
academic and industrial experience handling, expressing, developing, and commercializing
cytochrome P450 enzymes.
Mechanistic Basis and Exploration of Alix Incorporation into Exosomes-Robert Piper, Professor, Department of Molecular Physiology and Biophysics $35,000
Collaborator: Michael Henry, Professor, Department of Molecular Physiology and Biophysics
Executive Summary
Many cells release small membrane vesicles that contain a variety of protein and RNA
components. There has been an intense interest in these ‘exosomes’ over the last decade
due to their potential as biomarkers as well as vehicles to deliver components to different
compartments in the body. Yet the molecular basis for how specific proteins are selected for
incorporation into exosomes, and the technology to use such knowledge to effectively program
biologically-engineered exosomes with a particular content, remain elusive. This proposal
aims to provide such mechanistic insight and to use that to build chimeric molecules that can
package proteins and RNAs into exosomes. These experiments will provide the preliminary
data for extramural funding to better explore the biological mechanisms behind exosome
formation in conjunction with studies on how to load exosomes with specific content and to
target exosomes to particular tissues.
Targeting RAD51 DNA repair protein for cancer therapy: development of a combined experimental/computational workflow- Maria Spies, Professor, Department of Pharmaceutical Sciences and Experimental Therapeutics $35,000
Collabortor: M Ashely Spies, Professor, Department of Pharmaceutical Sciences and Experimental Therapeutics
Excutive Summary
We are seeking support to initiate a drug-discovery campaign targeting human RAD51
recombinase, an important player in homology-directed DNA repair. Several campaigns targeting
RAD51 demonstrated a therapeutic potential, but yielded compounds with poor drug-like
properties. This seed grant will allow us to carry out a fragment screen to identify RAD51 binders
by SPR and to characterize the effect of the hits on the biochemical activities of RAD51. We are
in a unique position to overcome multiple challenges associated with targeting RAD51 as we
possess a high quality model of the RAD51 nucleoprotein filament, a strategy to surface-tether a
monomeric form of RAD51, an experimental and computational pipeline for growing and
combining the fragments and scaffold hopping, and a set of biochemical assays to determine the
effect of the identified small molecules. The results of these studies will constitute essential
preliminary data for submission of a multi-PI R01 and/or SBIR grant.
Initial Seed Grants Awardees- August 27, 2018-August 26, 2019
Amyloid Proteins with a Role in Animal Development- Jan Fassler, Professor, Department of Biology $35,000
Collaborators: Bryan Phillips, Associate Professor, Department of Biology; Daniel Weeks, Professor, Department of Biochemistry
Executive Summary
Amyloid formation and accumulation is central to many neurodegenerative diseases, however,
paradoxically, mechanisms that generate amyloid aggregates are well-conserved across species. The
presence of amyloids in frog oocytes and worm embryos led us to hypothesize a requirement for their
formation and regulated processing during early animal development. Our immediate goal is to
understand the developmental role of amyloids, and, long-term, identify their dedicated disaggregases.
Here, we propose a genetic screen to identify proteins whose amyloid conformation is necessary for
normal C. elegans development. The intrinsic aggregating propensity of each candidate will be
evaluated using GFP-tagged protein fusions in C. elegans embryos, and in the amyloid-rich nuclear
environment of Xenopus oocytes, as well as in yeast and in vitro. These experiments are significant
because they will add a novel layer of protein regulation to the arsenal of gene regulatory networks
already recognized as important during animal development
Harnessing gut microbiome-mediated metabolism of the natural isoflavone daidzein to treat breast cancer - Edward Filardo, Professor, Surgical Oncology $35,000
Collaborator: Ashutosh Mangalam, Professor in Surgery, Department of Pathology
Executive Summary
Blockade of nonovarian estrogen is the most widely administered and well-tolerated cancer therapy.
Dietary estrogens, such as the natural soy isoflavone, daidzein are abundant bioavailable estrogens
whose physiological concentration exceeds endogenous estrogens, and whose pathogenic potential is
determined solely by the gut microbiome. This proposal by Drs. Mangalam and Filardo is a crossdisciplinary
study spanning microbial pathogenesis, molecular endocrinology and pharmacology to
investigate the role of daidzein-metabolizing bacteria in breast cancer development. Here, funds are
requested for preliminary studies in human subjects, and in mice to generate publications and data to
support NCI-based grant awards. Initial federal grant applications will focus on employing humanderived,
daidzein-metabolizing, anaerobic gut bacteria (eg Slackia isoflavoniconvertens). Later
applications will represent further “proof of concept” by testing necessary and sufficient
contingencies of daidzein to equol metabolism using Escherichia coli K12 derivatives that separately
express the three bacterial enzymes involved in the daidzein to S-equol pathway.
Chemically modified RNA activated matrices for bone regeneration -Aliasger Salem, Professor, Department of Pharmaceutical Sciences and Experimental Therapeutics $35,000
Collaborator: Satheesh Elangovan, Professor, Department of Periodontics
Executive Summary
There is an ever increasing need for novel biomaterials mimicking the natural wound healing process
that can predictably enhance bone regeneration and favor normal fracture healing in patients.
Ribonucleic acids (RNA) when chemically modified at their bases (cmRNA) have the unique advantage
of functioning entirely in the cytoplasm (avoiding the need for nuclear entry) thereby leading to a
targeted transient expression of desired proteins with immense potential to overcome the challenges
that exist with protein and DNA based approaches. We propose to identify the best combination and
sequential delivery of cmRNA encoding key regenerative factors, which will then be evaluated for
efficacy in a calvarial bone defect animal model.