Genetically Modified Host Cells for Isoprenoid Production

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Solution Overview

Problem

Current methods for producing isoprenoids are limited by low yields due to the low accumulation of these compounds in natural sources, difficulties in large-scale cultivation, and the need for toxic solvents in extraction, making commercial production challenging.

Innovation Solution

Genetically modified host cells are developed to overexpress mevalonate pathway enzymes, particularly mevalonate kinase, to increase the production of isoprenoid compounds by balancing enzyme activity levels and using higher copy number plasmids and stronger promoters to enhance gene expression.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If isoprenoids are extracted from natural sources, then isoprenoid compounds can be obtained, but the yield is very low due to low accumulation in nature

Engineering Contradiction:

Improveisoprenoid yieldVSAvoidproduction efficiency

Core Design Contradiction:

Quantity of substanceVSProductivity

Solution Approach 1:

The patent uses genetically modified host cells that contain the complete mevalonate pathway enzymes, enabling the cells to self-produce isoprenoid compounds internally through metabolic engineering, eliminating the need for extraction from natural sources and dramatically increasing yield

Inventive Principle:

Principle #25Self-service

Solution Approach 2:

The patent modifies the metabolic parameters of host cells by introducing and overexpressing key mevalonate pathway enzymes (HMGS, HMGR, MK, PMK, MPD), changing the cellular metabolic state to favor high-level isoprenoid production

Inventive Principle:

Principle #35Parameter changes

2Quantity of substance

If natural sources are used for isoprenoid production, then isoprenoid compounds can be obtained, but large-scale cultivation is difficult

Engineering Contradiction:

Improveisoprenoid quantityVSAvoidcultivation difficulty

Core Design Contradiction:

Quantity of substanceVSEase of manufacture

Solution Approach 1:

The patent creates simplified cellular copies of the isoprenoid production system by engineering host cells to contain the necessary metabolic pathways, allowing production in controlled fermentation systems rather than requiring cultivation of complex natural sources

Inventive Principle:

Principle #26Copying

Solution Approach 2:

The patent uses genetically modified host cells as intermediaries that convert readily available carbon sources into isoprenoid compounds through the engineered mevalonate pathway, serving as a bridge between simple cultivation and complex isoprenoid production

Inventive Principle:

Principle #24Intermediary (Mediator)

3Quantity of substance

If extraction methods are used for isoprenoid production, then isoprenoid compounds can be obtained, but toxic solvents are required necessitating special handling

Engineering Contradiction:

Improveisoprenoid recoveryVSAvoidtoxic solvent handling

Core Design Contradiction:

Quantity of substanceVSObject-affected harmful factors

Solution Approach 1:

The patent replaces the mechanical/chemical extraction system with a biological production system where isoprenoids are synthesized within host cells and can be recovered through cell disruption and extraction with less toxic solvents, or through direct fermentation broth processing

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent converts the harm of low natural accumulation into a benefit by using genetic engineering to overload the host cell's metabolic pathways with isoprenoid production, transforming a limiting factor into a driving force for high-yield production

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

4Productivity

If mevalonate pathway enzymes are overexpressed, then isoprenoid production increases, but enzyme activity balance becomes challenging

Engineering Contradiction:

Improveisoprenoid production levelVSAvoidenzyme activity balancing

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The patent employs dynamic control of enzyme expression by using inducible promoters and varying induction conditions for different mevalonate pathway enzymes, allowing optimization of enzyme activity balance throughout the fermentation process

Inventive Principle:

Principle #15Dynamics

Solution Approach 2:

The patent uses varying copy numbers of expression vectors for different enzymes, providing excessive expression of rate-limiting enzymes while maintaining appropriate levels of other pathway enzymes, thereby achieving overall pathway optimization

Inventive Principle:

Principle #16Partial or excessive action

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

This approach significantly increases isoprenoid production levels, enabling the production of higher quantities than current technologies, reducing the reliance on toxic solvents and improving commercial viability.

Implementation Method 1

microbes have been engineered to overexpress a part of or the entire mevalonate pathway for production of the isoprenoid amorpha-4,11-diene

Methodology Applied

Scientific EffectMevalonate pathway: Fermentation

Data Source

PatentEP3020799B1Production of isoprenoids and isoprenoid precursors

Publication Date: 2020.08.26 AMYRIS BIOTECHNOLOGIES INC (US)

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AI summary

The present invention provides genetically modified host cells and use of same for producing isoprenoid compounds.