Nepeta cataria L. (catnip) can serve as a chassis for the engineering of secondary metabolic pathways

Objective Evaluation of Nepeta cataria as a host with specific endogenous metabolite background for transient expression and metabolic engineering of secondary biosynthetic sequences. Results The reporter gene gfp::licBM3 as well as three biosynthetic genes leading to the formation of the cannabinoi...

Full description

Saved in:
Bibliographic Details
Published in:Biotechnology letters 2024-10, Vol.46 (5), p.843-850
Main Authors: Geissler, Marcus, Neubauer, Christoph, Sheludko, Yuriy V., Brückner, Adrian, Warzecha, Heribert
Format: Article
Language:English
Subjects:
Applied Microbiology
Biochemistry
Biomedical and Life Sciences
Biotechnology
Cannabinoids
Cloning
Combinatorial analysis
Cultivars
Gene sequencing
Genes
Glycosides
Life Sciences
Metabolic engineering
Metabolic pathways
Metabolism
Metabolites
Microbiology
Modular engineering
Nepeta cataria
Original Research Paper
Reporter gene
Citations: Items that this one cites
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Objective Evaluation of Nepeta cataria as a host with specific endogenous metabolite background for transient expression and metabolic engineering of secondary biosynthetic sequences. Results The reporter gene gfp::licBM3 as well as three biosynthetic genes leading to the formation of the cannabinoid precursor olivetolic acid were adopted to the modular cloning standard GoldenBraid, transiently expressed in two chemotypes of N. cataria and compared to Nicotiana benthamiana . To estimate the expression efficiency in both hosts, quantification of the reporter activity was carried out with a sensitive and specific lichenase assay. While N.   benthamiana exhibited lichenase activity of 676 ± 94 μmol g −1  s −1 , N. cataria cultivar ‘1000’, and the cultivar ‘Citriodora’ showed an activity of 37 ± 8 μmol g −1  s −1 and 18 ± 4 μmol g −1  s −1 , respectively. Further, combinatorial expression of genes involved in cannabinoid biosynthetic pathway acyl-activating enzyme 1 ( aae1 ), olivetol synthase ( ols ) and olivetolic acid cyclase ( oac ) in N. cataria cv. resulted presumably in the in vivo production of olivetolic acid glycosides. Conclusion Nepeta cataria is amenable to Agrobacterium -mediated transient expression and could serve as a novel chassis for the engineering of secondary metabolic pathways and transient evaluation of heterologous genes.
ISSN:0141-5492
1573-6776
1573-6776
DOI:10.1007/s10529-024-03489-w