Cloning, functional characterization and evaluating potential in metabolic engineering for lavender ( +)-bornyl...

Area of Science:

• Plant biochemistry
• Monoterpenoid biosynthesis
• Enzyme characterization

Background:

• (+)-Borneol and camphor are key monoterpenoids contributing to plant aroma and medicinal value.
• (+)-Borneol is synthesized from geranyl diphosphate (GPP) via (+)-bornyl diphosphate (BPP) by (+)-bornyl diphosphate synthase ((+)-BPPS).
• Understanding the function of (+)-BPPS is crucial for manipulating these valuable compounds in plants.

Purpose of the Study:

• To isolate and functionally characterize a novel (+)-bornyl diphosphate synthase ((+)-LiBPPS) from Lavandula x intermedia.
• To investigate the in planta role of (+)-LiBPPS in monoterpenoid metabolism.
• To assess the potential of (+)-LiBPPS in biotechnological applications for borneol and camphor production or reduction.

Main Methods:

• Isolation and cloning of the (+)-LiBPPS cDNA from Lavandula x intermedia.
• Expression of recombinant (+)-LiBPPS in E. coli and protein purification.
• Functional characterization of the recombinant enzyme using GPP as a substrate.
• Generation of sense and antisense transgenic Lavandula latifolia plants to study in planta function.

Main Results:

• The recombinant (+)-LiBPPS successfully catalyzed the conversion of GPP to BPP, with BPP as the major product.
• Antisense overexpression of (+)-LiBPPS in planta led to decreased production of (+)-borneol and camphor without affecting plant growth.
• Sense overexpression of (+)-LiBPPS enhanced borneol and camphor production but severely impaired plant growth and development.

Conclusions:

• (+)-LiBPPS plays a significant role in the biosynthesis of (+)-borneol and camphor in plants.
• The enzyme is a potential target for metabolic engineering to control borneol and camphor levels in plants.
• Targeted expression strategies, such as using trichome-specific promoters, may be necessary for successful sense overexpression applications.