Characterization of Three Types of Elongases from Different Fungi and Site-Directed Mutagenesis

Author:

Wang Yuxin12,Chang Lulu12,Zhang Hao123,Chen Yong Q.12ORCID,Chen Wei123,Chen Haiqin12

Affiliation:

1. State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi 214122, China

2. School of Food Science and Technology, Jiangnan University, Wuxi 214122, China

3. National Engineering Research Center for Functional Food, Jiangnan University, Wuxi 214122, China

Abstract

Fatty acid elongases play crucial roles in synthesizing long-chain polyunsaturated fatty acids. Identifying more efficient elongases is essential for enhancing oleaginous microorganisms to produce high yields of target products. We characterized three elongases that were identified with distinct specificities: McELO from Mucor circinelloides, PrELO from Phytophthora ramorum, and PsELO from Phytophthora sojae. Heterologous expression in Saccharomyces cerevisiae showed that McELO preferentially elongates C16 to C18 fatty acids, PrELO targets Δ6 polyunsaturated fatty acids, and PsELO uses long chain saturated fatty acids as substrates. McELO and PrELO exhibited more homology, potentially enabling fatty acid composition remodeling and enhanced LC-PUFAs production in oleaginous microorganisms. Site-directed mutagenesis of conserved amino acids across elongase types identified residues essential for activity, supported by molecular docking. Alanine substitution of conserved polar residues led to enzyme inactivation, underscoring their importance in the condensation reaction. Our findings offer promising elongase candidates for polyunsaturated fatty acid production, contributing to the bioindustry’s sustainable development.

Funder

National Key Research and Development Program of China

China Postdoctoral Science Foundation Funded Project

Program of “Collaborative Innovation Center for Food Safety and Quality Control in Jiangsu Province”

Publisher

MDPI AG

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