# Acquisition of a Preparative High-Performance Liquid Chromatography (HPLC) System

> **NIH NIH R01** · UNIVERSITY OF OKLAHOMA · 2024 · $105,000

## Abstract

Project Summary of the Funded Parent Award (1R01GM138800-01A1). Isoprenoids represent a
diverse class of compounds with a broad range of applications in medicine and industry. Their extraction
from natural sources is both challenging and potentially harmful to the environment, while the enormous
structural complexity of many isoprenoids makes traditional chemical synthesis nontrivial. Modern
metabolic engineering and synthetic biology approaches have overcome some of these difficulties, but
issues related to metabolic flux and the limited availability of the universal isoprenoid precursors complicate
their widespread implementation. The artificial pathways developed thus far have been solely focused on
synthesizing dimethylallyl and isopentenyl diphosphates and require additional enzymes for the generation
of polyprenyl-diphosphates (polyprenyl-PPs). Thus, the primary objective of this proposal is to develop an
efficient strategy for the synthesis of both natural and unnatural (poly)prenyl-PPs for downstream
applications. This will be achieved using two complementary methods: i) employing undecaprenol kinases
and isopentenyl phosphate kinases; and ii) employing hydroxyethylthiazole kinase, isopentenyl phosphate
kinases, and farnesyl diphosphate synthase. Additionally, the two methods will work in conjunction with
isoprenoid methylatransferases to incorporate additional diversity into the polyprenyl-PPs. The proposed
studies include: i) structural and functional assessment of selected enzymes, ii) catalyst engineering, and
iii) optimization of coupled in vitro and in vivo platforms for the generation of diversified libraries of select
natural products. We expect these studies to generate: i) rules and concepts to advance knowledge on
structure-activity relationships in selected classes of enzymes; ii) an optimized, enzyme-coupled platform
to generate diversified substrates and isoprenoids; and iii) novel isoprenoid analogs with potential
therapeutic applications. Thus, the proposed work will offer unprecedented access to uniquely bioactive
isoprenoid libraries not readily accessible via traditional methods, and it stands to deepen our fundamental
understanding of four enzyme classes while also developing them into useful biocatalysts.

## Key facts

- **NIH application ID:** 11096187
- **Project number:** 3R01GM138800-03S1
- **Recipient organization:** UNIVERSITY OF OKLAHOMA
- **Principal Investigator:** Shanteri Singh
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $105,000
- **Award type:** 3
- **Project period:** 2022-01-01 → 2026-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11096187

## Citation

> US National Institutes of Health, RePORTER application 11096187, Acquisition of a Preparative High-Performance Liquid Chromatography (HPLC) System (3R01GM138800-03S1). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/11096187. Licensed CC0.

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