# In Vitro Glycorandomization of Natural Products

> **NIH NIH R37** · UNIVERSITY OF KENTUCKY · 2021 · $445,994

## Abstract

ABSTRACT OF THE RESEARCH PLAN
 The studies to be pursued under the proposed extension fall within three general areas: i) enzyme catalyst
engineering and modulation (theme 1); ii) selected platform applications (theme 2); and iii) a systematic study
of the impact of glycosylation on fundamental drug properties (theme 3). The key tenets of each are briefly
described below and parallel the consistently high level of significance and innovation reflected in past periods
of support. The overarching significance stems from the anticipated advancements of universal principles for
enzyme catalyst engineering/modulation, novel enabling strategies for synthesis and biosynthetic study, unique
probes or leads of significance to diverse therapeutic areas, and a potential new conceptual framework for the
general use of glycosylation in drug development. Anticipated drivers of innovation are both by design
(particularly novel synthetic and assay strategies) and directed discovery (unprecedented glycosyltransferase
modulators and probes/leads with high potential to lead to new understanding in the fundamentals of
immunomodulation and/or inflammation).
 Studies directed toward enzyme catalyst engineering and modulation (theme 1) will focus on comparing the
directed evolutionary paths among the structurally similar glycosyltransferases (GTs) OleD and OleI, assessing
the transferability of beneficial mutations across OleD and OleI and identifying mutational hot spots within the
atypical GTB-fold GT AmphD1. The comparative OleD/OleI studies will consist of: i) the evaluation of an ‘OleI
Loki,’ which contains the 5 key mutations of structurally conserved residues that led to OleD Loki; and ii) a
parallel epPCR-directed evolution approach for increasing OleI proficiency/permissivity using the same screens
and mutational strategy that led to the discovery of enhanced OleD progeny. Key outcomes of this comparative
study will be a potential universal blueprint for improving the proficiency/promiscuity of most GTB-fold GTs and
improved GTs for select sugar-conjugation (D-Gal analogs) to support theme 3. The directed evolution of
AmphD1 (and parallel proposed structure elucidation and structure-based engineering) will also follow our OleD
precedent and offers both the potential to expand the universal GT engineering blueprint toward atypical GTs
and to identify GT variants to dramatically simplify the targeted synthesis of the unique preclinical antifungal lead
2’-epi-amphotericin (theme 2). In addition, we will pursue the mechanistic study of small molecule inhibitors and
activators identified via the ClNP-Glc-based HT screen - studies that hold high potential in the discovery of new
paradigms for GT catalytic modulation that may extend to other enzyme families and/or have pharmacologic
applications. Finally, while we remain enthusiastic about the uncharted frontier of enzyme modulation via
controlled electrical fields, such technically challenging experiments require th...

## Key facts

- **NIH application ID:** 10158383
- **Project number:** 5R37AI052218-20
- **Recipient organization:** UNIVERSITY OF KENTUCKY
- **Principal Investigator:** Jon Scott Thorson
- **Activity code:** R37 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $445,994
- **Award type:** 5
- **Project period:** 2002-06-03 → 2024-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10158383

## Citation

> US National Institutes of Health, RePORTER application 10158383, In Vitro Glycorandomization of Natural Products (5R37AI052218-20). Retrieved via AI Analytics 2026-06-12 from https://api.ai-analytics.org/grant/nih/10158383. Licensed CC0.

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