# Understanding Oligomer Assembly of Isoprenoid and Terpene Synthases

> **NIH NIH F32** · UNIVERSITY OF PENNSYLVANIA · 2022 · $10,651

## Abstract

Project Summary/Abstract
 Oligomeric assembly of terpene synthases improves biosynthetic product flux by proximity or cluster
channeling. The unusual diterpene (C20) synthase, PvCPS, from Penicillium verruculosum was identified and
characterized as the first bifunctional terpene synthase with abg domain architecture containing both
prenyltransferase and class II cyclase activities. The C-terminal a domain generates geranylgeranyl diphosphate
which is then cyclized to copalyl diphosphate at the interface of the N-terminal bg domains. Upon recombinant
expression and purification, I determined that PvCPS exists as a hexamer at high concentrations – a unique
quaternary structure for known abg terpene synthases – and dissociates to monomers at low concentrations.
 Interestingly, oligomerization is common among prenyltransferases and terpene synthases, which have
been observed as monomers, dimers, trimers, tetramers, and hexamers. The first part of this proposal aims to
determine the structural aspects of oligomer formation using the prenyltransferase-cyclase chimera of PvCPS
by determining the high-resolution structure using Cryo-Electron Microscopy (Cryo-EM). The architecture of
PvCPS presents an ideal engineering opportunity for multifunctional assembly-line terpene biosynthesis. Thus,
the second part of this proposal aims to utilize the oligomer assembly of PvCPS to engineer novel multifunctional
enzymes, since many terpene products exhibit useful pharmaceutical properties. Successful completion of these
aims will lead to the development of multifunctional synthases for in vitro biosynthesis of complex terpenes.
 Lastly, human prenyltransferases, such as farnesyl diphosphate synthase (hFPPS), geranylgeranyl
diphosphate synthase (hGGPPS), and squalene synthase (hSQS), have been implicated in a variety of diseases
and cancers. Each of these prenyltransferases has been characterized as an oligomer in vitro. However, if
oligomer assembly is concentration-dependent, does oligomer assembly occur under cellular conditions? And if
so, what is its role? The last aim will determine if oligomer assembly occurs in vivo, and its associated function.

## Key facts

- **NIH application ID:** 10328886
- **Project number:** 5F32GM137461-02
- **Recipient organization:** UNIVERSITY OF PENNSYLVANIA
- **Principal Investigator:** Trey Adam Ronnebaum
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $10,651
- **Award type:** 5
- **Project period:** 2021-03-01 → 2022-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10328886

## Citation

> US National Institutes of Health, RePORTER application 10328886, Understanding Oligomer Assembly of Isoprenoid and Terpene Synthases (5F32GM137461-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10328886. Licensed CC0.

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