Fungal Host Genome Modification for Fermentation

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Solution Overview

Problem

Microbial fermentation processes face challenges such as reduced yield and increased costs due to issues like viscosity, unwanted by-products, and mycotoxin production, which hinder standardization of fermentation and downstream processing protocols, particularly when using fungi as host systems.

Innovation Solution

Modifying the genome of microbial host cells to deficiency in non-ribosomal peptide synthase production, using techniques like gene replacement and deletion, to reduce the production of insoluble by-products and improve fermentation conditions, thereby enhancing yield and purity of compounds.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If fungal host systems are used for microbial fermentation, then productivity and versatility of compound production are improved, but harmful factors increase due to mycotoxin production and unwanted by-products

Engineering Contradiction:
Improvecompound productionVSAvoidmycotoxin production
Core Design Contradiction:
ProductivityVSObject-generated harmful factors

Solution Approach 1:

The patent removes harmful NRPS genes from the fungal genome through targeted deletion. Specifically, genes encoding non-ribosomal peptide synthases (NRPS) are identified and deleted using molecular biology techniques, thereby extracting the harmful toxin-producing capability while preserving the beneficial metabolic pathways for compound production.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent modifies genetic parameters by deleting specific NRPS genes (such as npsA, npsB, npsC, npsD, npsE) to change the biochemical output profile of the fungus. This genetic parameter change eliminates mycotoxin production while maintaining or enhancing production of desired compounds through altered metabolic flux.

Inventive Principle:
Principle #35Parameter changes

2Productivity

If fungal host systems are used for microbial fermentation, then productivity is improved, but downstream processing difficulty increases due to insoluble by-products and coating of fermentation devices

Engineering Contradiction:
Improvecompound productionVSAvoiddownstream processing
Core Design Contradiction:
ProductivityVSEase of manufacture

Solution Approach 1:

The patent extracts and removes the genetic basis for producing insoluble by-products by deleting NRPS genes. This eliminates the formation of hydrophobins and other insoluble peptides that cause filtration problems and device coating, thereby simplifying downstream processing while maintaining high productivity.

Inventive Principle:
Principle #2Taking out (Extraction)

3Manufacturing precision

If fermentation conditions are optimized for each specific product, then manufacturing precision is improved, but device complexity and loss of time increase due to lack of standardization

Engineering Contradiction:
Improvefermentation optimizationVSAvoidprotocol standardization
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent creates a universal fungal host strain with deleted NRPS genes that can be used for producing multiple different compounds under standardized conditions. This multi-functional host eliminates the need for separate optimization protocols for each product, as the modified fungus consistently produces fewer harmful by-products across different fermentation processes.

Inventive Principle:
Principle #6Universality (Multi-functionality)

Data Source

PatentEP2588616B1A method for the production of a compound of interest
Publication Date: 2018.11.14 DSM IP ASSETS BV
  • EP2588616B1 patent drawingFigure 1
  • EP2588616B1 patent drawingFigure 2(1)~2(4)
  • EP2588616B1 patent drawingFigure 3(1)~3(4)

AI summary

The present invention relates to a method for the production of a compound of interest by microbial fermentation, wherein the microbial host cell used has been modified in its genome such that it results in a deficiency in the production of at least one non-ribosomal peptide synthase. The present invention further relates to a microbial host cell that has been modified in its genome such that it results in a deficiency in the production of at least one non-ribosomal peptide synthase. The invention further relates to a compound of interest.