Identification of novel isoprene synthases through genome mining and expression in Escherichia coli

Isoprene is a naturally produced hydrocarbon emitted into the atmosphere by green plants. It is also a constituent of synthetic rubber and a potential biofuel. Microbial production of isoprene can become a sustainable alternative to the prevailing chemical production of isoprene from petroleum. In t...

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Bibliographic Details
Published in:Metabolic engineering 2015-09, Vol.31, p.153-162
Main Authors: Ilmén, Marja, Oja, Merja, Huuskonen, Anne, Lee, Sangmin, Ruohonen, Laura, Jung, Simon
Format: Article
Language:English
Subjects:
Alkyl and Aryl Transferases - chemistry
Alkyl and Aryl Transferases - isolation & purification
Alkyl and Aryl Transferases - physiology
Amino Acid Sequence
Butadienes
Escherichia coli - genetics
Genome mining
Genome, Bacterial
Hemiterpenes - biosynthesis
Homology-based screening
Isoprenoid
Molecular Sequence Data
Pentanes
Sequence Homology
Terpene
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Summary:Isoprene is a naturally produced hydrocarbon emitted into the atmosphere by green plants. It is also a constituent of synthetic rubber and a potential biofuel. Microbial production of isoprene can become a sustainable alternative to the prevailing chemical production of isoprene from petroleum. In this work, sequence homology searches were conducted to find novel isoprene synthases. Candidate sequences were functionally expressed in Escherichia coli and the desired enzymes were identified based on an isoprene production assay. The activity of three enzymes was shown for the first time: expression of the candidate genes from Ipomoea batatas, Mangifera indica, and Elaeocarpus photiniifolius resulted in isoprene formation. The Ipomoea batatas isoprene synthase produced the highest amounts of isoprene in all experiments, exceeding the isoprene levels obtained by the previously known Populus alba and Pueraria montana isoprene synthases that were studied in parallel as controls. •Three uncharacterized terpene synthases were shown to code for active isoprene synthases.•Production of isoprene by the novel and previously known isoprene synthases was assessed in E. coli.•The newly identified enzyme from Ipomoea batatas produced isoprene effectively.•batatas IspS is an attractive option for development of microbial isoprene factories.
ISSN:1096-7176
1096-7184
DOI:10.1016/j.ymben.2015.08.001