Lanosterol Synthase Engineering for Isoprenoid Yield
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Solution Overview
Problem
The production of isoprenoids and their precursors from natural sources is hindered by the structural diversity of isoprenoids, leading to high production costs and low yields, and the mevalonate pathway used by eukaryotes and other organisms is limited by bottlenecks and off-target compound production.
Innovation Solution
Engineering host cells with heterologous polynucleotides encoding hypomorphic lanosterol synthases and squalene epoxidases, which reduce the activity of these enzymes, increases the production of isoprenoids and their precursors by altering the flux through the mevalonate pathway.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If wild-type lanosterol synthase is used in the mevalonate pathway, then the pathway proceeds through normal metabolic flux, but the production of isoprenoids and precursors is limited due to pathway bottlenecks and off-target compound production
Solution Approach 1:
The patent applies parameter changes by introducing amino acid substitutions at specific positions (e.g., E287G, K329N, E617V, F726L) in the lanosterol synthase enzyme to alter its catalytic properties. These parameter changes in enzyme structure modify the reaction pathway to reduce off-target compounds while enhancing isoprenoid production, directly resolving the technical contradiction between productivity and harmful byproducts.
2Adaptability or versatility
If structural diversity of isoprenoids is utilized for various applications, then the versatility of isoprenoid compounds increases, but the extraction yield from natural sources decreases and production costs increase
Solution Approach 1:
The patent uses parameter changes by engineering lanosterol synthase with specific amino acid substitutions to redirect metabolic flux toward desired isoprenoid products. This enables high-yield production of diverse isoprenoids in recombinant host cells, maintaining versatility while dramatically improving productivity compared to natural extraction methods.
Solution Approach 2:
The patent employs an intermediary approach by using recombinant host cells expressing engineered lanosterol synthase as a biological factory. This intermediary system converts simple precursors into diverse isoprenoids through controlled enzymatic reactions, bypassing the need for complex natural source extraction while maintaining product versatility.
3Ease of manufacture
If mevalonate pathway is used for isoprenoid synthesis, then the pathway provides a established biosynthetic route, but the pathway efficiency is limited by bottlenecks and off-target compound production
Solution Approach 1:
The patent applies parameter changes by modifying key enzyme parameters in the mevalonate pathway, specifically lanosterol synthase amino acid sequences. These changes optimize catalytic efficiency and substrate specificity, removing pathway bottlenecks while maintaining the ease of manufacture provided by the established mevalonate pathway framework.
Data Source
AI summary
Described in this application are proteins and host cells involved in methods of producing isoprenoid precursors and/or isoprenoids.