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Evolution and Distribution of C7-Cyclitol Synthases in Prokaryotes and Eukaryotes
Andrew R Osborn1, Kelsey M Kean2, Khaled M Alseud1
1Department of Pharmaceutical Sciences, Oregon State University , Corvallis, Oregon 97331-3507, United States.
ACS Chemical Biology
|February 10, 2017
Summary
2-Epi-5-epi-valiolone synthase (EEVS) and desmethyl-4-deoxygadusol synthase (DDGS) are C7-sugar phosphate cyclases with broad distribution. This study clarifies their identification, structure, and function, revealing their widespread biological roles.
Area of Science:
- Biochemistry
- Enzymology
- Genomics
Background:
- 2-Epi-5-epi-valiolone synthase (EEVS) and desmethyl-4-deoxygadusol synthase (DDGS) are C7-sugar phosphate cyclases (SPCs) with homologous structures to 3-dehydroquinate synthase (DHQS).
- EEVS is involved in C7N-aminocyclitol biosynthesis, while DDGS is crucial for mycosporine-like amino acid sunscreen production.
- Current annotations inaccurately classify these enzymes, hindering research.
Purpose of the Study:
- To develop reliable methods for distinguishing EEVS and DDGS enzymes.
- To elucidate the structural and functional characteristics of EEVS and DDGS.
- To investigate the evolutionary distribution and potential roles of EEVS and DDGS genes across diverse organisms.
Main Methods:
- Bioinformatic analysis of genome sequences to identify conserved sequence features.
- X-ray crystallography to determine the structure of a representative DDGS enzyme.
- Site-directed mutagenesis to probe the function of active site residues.
- Functional characterization of novel EEVS enzyme clade representatives.
Main Results:
- Identified key sequence features for accurate EEVS and DDGS classification.
- Reported a crystal structure of DDGS, highlighting high similarity and distinct active site residues compared to EEVS.
- Demonstrated the catalytic importance of specific active site residues through mutagenesis.
- Confirmed the identity and function of novel EEVS enzymes from a distinct evolutionary clade.
- Documented the widespread distribution of EEVS and DDGS genes in prokaryotes and eukaryotes.
Conclusions:
- Accurate identification and characterization of EEVS and DDGS are crucial for understanding their roles in natural product biosynthesis.
- Structural and functional insights reveal conserved and divergent features of these SPCs.
- The broad phylogenetic distribution suggests significant, yet largely unexplored, biological functions across diverse life forms.
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