Fungal Terpene Synthase Engineering for Selective Production
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Solution Overview
Problem
The study of fungal terpene synthases has been lagging behind plant terpene synthases, with limited characterization and understanding of their diversity and potential for producing novel terpenoids, which hampers the industrial biotechnology sector's ability to harness their commercial value due to insufficient activity and selectivity of existing terpene synthases.
Innovation Solution
A bacterial strain is engineered to express fungal terpene synthases and genetically modified 1-deoxyxylulose-5-phosphate synthase enzymes, enhancing the production of terpenoids by improving enzyme activity and selectivity through specific amino acid mutations, and utilizing these strains to produce terpenoids in an expression medium.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If fungal terpene synthases are used for terpenoid production, then diversity of terpenoid products is improved, but enzyme activity and selectivity are insufficient
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid mutations at conserved positions in the fungal terpene synthase enzyme sequence. These mutations modify the enzyme's catalytic properties to enhance its activity and selectivity for producing specific terpenoid products, while maintaining the enzyme's ability to generate diverse terpene structures from common precursors.
2Ease of manufacture
If existing terpene synthases are used for industrial production, then production process is simple, but productivity is limited by insufficient enzyme activity
Solution Approach 1:
The patent modifies the enzyme's amino acid sequence parameters through targeted mutations to significantly enhance catalytic activity. This increases the productivity of terpenoid production while maintaining the simplicity of the overall production process, as the modified enzyme can be expressed in standard host systems without requiring complex process modifications.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The approach allows for the efficient production of diverse terpenoids with improved activity and selectivity, tapping into the vast untapped potential of fungal terpenoids and terpene synthases, thereby enhancing industrial biotechnology applications.
Implementation Method 1
terpene synthases (TPSs), which convert acyclic prenym diphosphate precursors into a multitude of cyclic and acyclic terpene scaffolds
Implementation Method 2
the ability of many TPSs to catalyze multiple terpene products from a single substrate
Implementation Method 3
genetically modified 1-deoxyxylulose-5-phosphate synthase (DXS) enzyme
Data Source
AI summary
A bacterial strain comprising one or more vectors encoding a) one or more enzymes to produce one or more terpene precursors; and b) a fungal terpene synthase (FTPS). The present invention also relates to a method of producing a terpenoid comprising a) culturing the bacterial strain described herein in an expression medium; and b) isolating the terpenoid from said expression medium.