Parallel Seed Train for High-Density Enzyme Inoculum

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Solution Overview

Problem

The conventional seed train process for producing ethanol from biomass is inefficient due to high costs, vulnerability to contamination, operator errors, and lengthy processing times, primarily because it requires multiple manual manipulations and starts with low cell densities, leading to increased production times and reduced productivity.

Innovation Solution

An improved seed train process where a single inoculum bioreactor is used to inoculate multiple production bioreactors, allowing for continuous culture and fermentation, thereby reducing the need for manual manipulations and increasing cell density more efficiently, thus optimizing the production of fermentation products like enzymes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If multiple successive propagations into progressively larger culture vessels are used, then sufficient microbial biomass is generated, but the process becomes time-consuming and vulnerable to contamination

Engineering Contradiction:
Improvemicrobial biomassVSAvoidprocess time
Core Design Contradiction:
Quantity of substanceVSLoss of time

Solution Approach 1:

The conventional single sequential seed train is segmented into multiple parallel seed trains. Each seed train starts from the same cryopreserved cell bank vial and progresses through identical culture vessels (shake flasks, spinners, wave bags, stirred bioreactors) simultaneously, rather than sequentially. This parallelization maintains the necessary biomass generation steps while eliminating the cumulative time delay of sequential processing.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

Multiple seed trains are initiated simultaneously from the cryopreserved cell bank vial at the beginning of the process, rather than waiting for each sequential train to complete. This preliminary parallel action ensures that all necessary biomass expansion steps occur concurrently, dramatically reducing total process time while still achieving sufficient inoculum density for production bioreactors.

Inventive Principle:
Principle #10Preliminary action

2Quantity of substance

If multiple manual manipulations are performed during each step, then culture propagation is achieved, but contamination risk and operator error increase

Engineering Contradiction:
Improvecell densityVSAvoidcontamination vulnerability
Core Design Contradiction:
Quantity of substanceVSReliability

Solution Approach 1:

The system is designed to minimize manual intervention by automating the propagation process. Once the parallel seed trains are initiated from the cryopreserved vials, the cultures propagate through all stages with minimal human contact. The system essentially serves itself by maintaining controlled environmental conditions and automated transfer mechanisms, reducing the opportunities for contamination and operator errors while still achieving high cell densities.

Inventive Principle:
Principle #25Self-service

3Ease of operation

If large-scale production bioreactors are started with low cell densities, then inoculation is simplified, but a lengthy 5-10 day growth phase is required

Engineering Contradiction:
Improveinoculation simplicityVSAvoidproduction time
Core Design Contradiction:
Ease of operationVSProductivity

Solution Approach 1:

The inoculum parameters (cell density and viability) are dramatically improved through the parallel seed train process. Instead of using low-density inocula that require long growth phases, the optimized process produces high-density, high-viability inoculum that can be directly inoculated into production bioreactors. This parameter change in the inoculum quality eliminates the need for lengthy 5-10 day growth phases while maintaining ease of inoculation operation.

Inventive Principle:
Principle #35Parameter changes

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 process enhances productivity and reduces costs by minimizing contamination risks, shortening production times, and achieving higher cell densities, leading to more efficient enzyme production and improved process control.

Implementation Method 1

preparing a fungal inoculum comprising fungal cells in an inoculum bioreactor

Methodology Applied
Scientific EffectFermentation: Fermentation

Implementation Method 2

culturing the fungal cells in the first production bioreactor to produce the fermentation product

Methodology Applied
Scientific EffectFermentation: Fermentation

Data Source

PatentUS11142783B2Seed train for large scale enzyme production
Publication Date: 2021.10.12 DSM IP ASSETS BV

AI summary

The invention relates to an optimized seed train expansion process.