Fern Albicanyl Diphosphate Synthase for Cost-Effective Albicanol Production
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Solution Overview
Problem
Current methods for producing sesquiterpenes like albicanol are complex and not cost-effective, with natural sources having low content, necessitating the discovery of new terpene synthases and more efficient production methods for valuable perfumery ingredients.
Innovation Solution
Identification of a novel albicanyl diphosphate synthase gene and its variants from the fern Dryopteris fragrans, which exhibit high selectivity in producing albicanyl diphosphate from FPP, utilizing a modified class I and class II motif, and classified as Haloacid dehalogenase-like hydrolase, enabling efficient biosynthetic production of albicanol.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If chemical synthesis approaches are used to produce sesquiterpenes, then production capability is achieved, but the process becomes complex and not cost-effective
Solution Approach 1:
The patent replaces complex chemical synthesis processes with a biochemical approach using terpene synthase enzymes. The enzyme-catalyzed conversion of FPP to albicanyl diphosphate and subsequent hydrolysis provides a simpler, more cost-effective pathway compared to traditional multi-step chemical synthesis, directly addressing the contradiction between production capability and process complexity
Solution Approach 2:
The patent introduces terpene synthase enzymes as intermediary biological catalysts to mediate the conversion of farnesyl diphosphate to albicanol. This enzymatic intermediary provides a controlled, selective, and efficient transformation pathway that avoids the complexity of direct chemical synthesis while maintaining high productivity
2Quantity of substance
If natural sources containing sesquiterpenes are used, then production is achieved, but the content of sesquiterpenes is low
Solution Approach 1:
The patent employs host cells (bacterial or plant) that are genetically engineered to express the terpene synthase enzyme and endogenously produce farnesyl diphosphate through their natural metabolism. The system serves itself by using the host's own metabolic pathways to generate substrate and the introduced enzyme to convert it to product, eliminating the need for external substrate addition and achieving high productivity
Solution Approach 2:
The patent performs preliminary genetic engineering of host cells to establish the complete biosynthetic pathway before production. The host cells are pre-modified to overexpress terpene synthase and optimize FPP availability, so that when cultivation begins, the system is already primed for high-yield sesquiterpene production, overcoming the low content limitation of natural sources
3Quantity of substance
If existing terpene synthases are used, then albicanol production is achieved, but selectivity is insufficient
Solution Approach 1:
The patent identifies and exploits specific local structural features within the terpene synthase enzyme - namely the class I motif (DDxxD) and class II motif (EXxHD) - that are responsible for substrate binding and catalytic selectivity. By focusing on these specific local regions with distinct amino acid sequences, the enzyme achieves high selectivity for albicanyl diphosphate production over other possible sesquiterpene products
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 novel synthase shows high selectivity and efficiency in producing albicanyl diphosphate, potentially leading to cost-effective and scalable production of albicanol, a valuable perfumery ingredient, by utilizing recombinant host cells like E. coli and transgenic plants.
Implementation Method 1
The albicanyl diphosphate synthase of the present invention and its variants may also be considered as 'haloacid-dehalogenase-like hydrolase'
Data Source
AI summary
Described herein is a method of producing a drimane sesquiterpene, such as an albicanol compound and/or derivatives thereof, by contacting at least one polypeptide with farnesyl diphosphate (FPP) with a polypeptide of the Haloacid dehalogenase-like (HAD-like) hydrolase superfamily as obtainable from plants of the genus Dryopteris, in particular of the species Dryopteris fragrans. The method may be performed in vitro or in vivo. Also described herein are amino acid sequences of polypeptides useful in the methods and nucleic acids encoding the polypeptides described. Also described herein are host cells or organisms genetically modified to express the polypeptides and useful to produce a drimane sesquiterpene such as an albicanol compound.