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Isopentenol Utilization Pathway for the Production of Linalool in Escherichia coli Using an Improved Bacterial Linalool/Nerolidol Synthase
- Source :
- Chembiochem, Ferraz, C A, Leferink, N G H, Kosov, I & Scrutton, N 2021, ' Isopentenol utilization pathway for the production of linalool in Escherichia coli using an improved bacterial linalool/nerolidol synthase ', CHEMBIOCHEM . https://doi.org/10.1002/cbic.202100110
- Publication Year :
- 2021
- Publisher :
- John Wiley and Sons Inc., 2021.
-
Abstract
- Linalool is a monoterpenoid used as a fragrance ingredient, and is a promising source for alternative fuels. Synthetic biology offers attractive alternative production methods compared to extraction from natural sources and chemical synthesis. Linalool/nerolidol synthase (bLinS) from Streptomyces clavuligerus is a bifunctional enzyme, producing linalool as well as the sesquiterpenoid nerolidol when expressed in engineered Escherichia coli harbouring a precursor terpenoid pathway such as the mevalonate (MVA) pathway. Here we identified two residues important for substrate selection by bLinS, L72 and V214, where the introduction of bulkier residues results in variants with reduced nerolidol formation. Terpenoid production using canonical precursor pathways is usually limited by numerous and highly regulated enzymatic steps. Here we compared the canonical MVA pathway to the non‐canonical isopentenol utilization (IU) pathway to produce linalool using the optimised bLinS variant. The IU pathway uses isoprenol and prenol to produce linalool in only five steps. Adjusting substrate, plasmid system, inducer concentration, and cell strain directs the flux towards monoterpenoids. Our integrated approach, combining enzyme engineering with flux control using the artificial IU pathway, resulted in high purity production of the commercially attractive monoterpenoid linalool, and will guide future efforts towards efficient optimisation of terpenoid production in engineered microbes.<br />Linalool is a monoterpenoid used as fragrance ingredient, and a promising source for alternative fuels. Here we explore the artificial isoprenol utilization pathway to produce linalool in Escherichia coli using the linalool/nerolidol synthase from Streptomyces clavuligerus. The generation of improved bLinS variants together with substrate and strain optimization increased linalool production and decreased nerolidol by‐product levels.
- Subjects :
- linalool
Protein Conformation
Acyclic Monoterpenes
Streptomyces clavuligerus
Mevalonic Acid
010402 general chemistry
medicine.disease_cause
01 natural sciences
Biochemistry
Prenol
chemistry.chemical_compound
Hemiterpenes
Pentanols
Linalool
terpenoids
Transferases
medicine
Escherichia coli
Amino Acid Sequence
Molecular Biology
Nerolidol
isopentenol utilization pathway
biology
Full Paper
010405 organic chemistry
Chemistry
Terpenes
Organic Chemistry
protein engineering
Isoprenol
Full Papers
biology.organism_classification
Terpenoid
Streptomyces
0104 chemical sciences
Molecular Medicine
synthetic biology
Flux (metabolism)
Sesquiterpenes
Signal Transduction
Subjects
Details
- Language :
- English
- ISSN :
- 14397633 and 14394227
- Volume :
- 22
- Issue :
- 13
- Database :
- OpenAIRE
- Journal :
- Chembiochem
- Accession number :
- edsair.doi.dedup.....2fa5951e2c6982580baadaeaa8be7c8a