MbtH-like Proteins Enhance Eukaryotic NRPS Secondary Metabolite Yield

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Solution Overview

Problem

Current industrial fermentation processes for secondary metabolites catalyzed by eukaryotic non-ribosomal peptide synthetases (NRPS) lack effective methods to enhance productivity, despite existing strategies, as MbtH-like proteins have only been shown to influence prokaryotic NRPS systems, leaving a gap in improving eukaryotic NRPS-dependent secondary metabolite production.

Innovation Solution

Contacting eukaryotic NRPS with MbtH-like proteins in eukaryotic hosts, specifically using fully eukaryotic NRPS systems that are not hybrids, to enhance the production of secondary metabolites by influencing NRPS-dependent intermediates and metabolites, with preferred MbtH-like proteins and host organisms like Penicillium and Aspergillus strains.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If MbtH-like proteins are used to enhance NRPS-dependent secondary metabolite production, then productivity is improved, but this approach has only been effective for prokaryotic NRPS systems, not eukaryotic ones

Engineering Contradiction:
Improvesecondary metabolite productionVSAvoidapplicability to eukaryotic NRPS systems
Core Design Contradiction:
ProductivityVSAdaptability or versatility

Solution Approach 1:

The patent applies universality by demonstrating that MbtH-like proteins, originally known to function only with prokaryotic NRPS systems, can also enhance eukaryotic NRPS-dependent secondary metabolite production. The invention shows that MbtH-like proteins from various sources (prokaryotic and eukaryotic) can universally improve productivity across different NRPS systems including eukaryotic ones, thereby expanding the applicability and multi-functionality of these proteins beyond their previously known prokaryotic context.

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

2Productivity

If conventional fermentation optimization strategies are applied, then some yield improvement is achieved, but further productivity enhancement remains limited

Engineering Contradiction:
Improveunit productivityVSAvoidprocess complexity
Core Design Contradiction:
ProductivityVSEase of manufacture

Solution Approach 1:

The patent applies parameter changes by introducing a new biological parameter (MbtH-like protein expression) into the fermentation system to enhance productivity. Rather than continuing to optimize conventional parameters like nutrient composition, pH, and temperature, the invention changes the system by adding MbtH-like proteins that interact with NRPS enzymes, thereby achieving further productivity improvement beyond what conventional optimization can provide.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent uses MbtH-like proteins as intermediaries that mediate between the NRPS enzyme system and the secondary metabolite production process. These proteins act as molecular mediators that enhance the activity of NRPS-dependent pathways, providing a bridge that connects existing cellular machinery to improved metabolite production without requiring complete pathway redesign.

Inventive Principle:
Principle #24Intermediary (Mediator)

3Productivity

If hybrid NRPS systems are used with MbtH-like proteins, then adenylation reaction is improved, but the system requires both eukaryotic and prokaryotic modules which complicates the system

Engineering Contradiction:
Improveadenylation reaction efficiencyVSAvoidNRPS system composition
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent applies the taking out principle by separating the MbtH-like protein function from the hybrid NRPS system complexity. The invention demonstrates that MbtH-like proteins can be added as independent components to enhance eukaryotic NRPS systems without requiring the creation of hybrid enzyme complexes. This extracts the beneficial MbtH function and applies it independently, avoiding the need to construct and maintain complex hybrid NRPS systems.

Inventive Principle:
Principle #2Taking out (Extraction)

Data Source

PatentEP3430153A1Mbth-like proteins in eukaryotic NRPS-catalyzed processes
Publication Date: 2019.01.23 CENTRIENT PHARMA NETHERLANDS BV

AI summary

The present invention relates to a method to improve the production of a secondary metabolite catalyzed by a non-ribosomal peptide synthetase comprising contacting in a eukaryotic host a eukaryotic non-ribosomal peptide synthetase with an MbtH-like protein. The present invention further relates to a composition comprising a eukaryotic non-ribosomal peptide synthetase that is not a hybrid and a prokaryotic MbtH and to a eukaryotic host cell comprising a non-ribosomal peptide synthetase and a polynucleotide allowing the expression of an MbtH-like protein.