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 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.
VTT Technical Research Centre of Finland Ltd., P.O. Box 1000, FI-02044 VTT, Espoo, Finland.
Marja
M
Ilmén
VTT Technical Research Centre of Finland Ltd., P.O. Box 1000, FI-02044 VTT, Espoo, Finland.
Merja
M
Oja
VTT Technical Research Centre of Finland Ltd., P.O. Box 1000, FI-02044 VTT, Espoo, Finland.
Anne
A
Huuskonen
Global Technology, SK Innovation, Daejeon, Republic of Korea.
Sangmin
S
Lee
VTT Technical Research Centre of Finland Ltd., P.O. Box 1000, FI-02044 VTT, Espoo, Finland.
Laura
L
Ruohonen
Global Technology, SK Innovation, Daejeon, Republic of Korea. Electronic address: jungmoongeun@sk.com.
Simon
S
Jung
D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000302:isolation & purification* / Q000502:physiology; D000595:Amino Acid Sequence; D002070:Butadienes; D004926:Escherichia coli / Q000235:genetics*; D016680:Genome, Bacterial; D045782:Hemiterpenes / Q000096:biosynthesis; D008969:Molecular Sequence Data; D010420:Pentanes; D017385:Sequence Homology
D002070:Butadienes; D045782:Hemiterpenes; D010420:Pentanes; C005059:isoprene; D019883:Alkyl and Aryl Transferases; C093854:isoprene synthase
Genome mining; Homology-based screening; Isoprenoid; Terpene