Mixed Sugar Fermentation Cell with Integrated Gene Copies

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Solution Overview

Problem

Current processes cannot efficiently convert glucose, arabinose, xylose, and galactose into fermentation products simultaneously, limiting the utilization of lignocellulosic biomass for renewable fuel production.

Innovation Solution

Development of a mixed sugar cell with integrated multiple copies of xylose isomerase and araA, araB, and araD genes, enabling the simultaneous conversion of glucose, xylose, arabinose, and galactose into fermentation products like ethanol.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If Saccharomyces cerevisiae is used for fermentation, then ethanol production is efficient, but C5 sugars (xylose and arabinose) cannot be utilized

Engineering Contradiction:
Improveethanol production efficiencyVSAvoidsugar utilization capability
Core Design Contradiction:
ProductivityVSAdaptability or versatility

Solution Approach 1:

The patent combines multiple gene functions into a single yeast strain by integrating xylose isomerase (xylA) and arabinose utilization genes (araA, araB, araD) into Saccharomyces cerevisiae. This merging of different metabolic capabilities allows the yeast to simultaneously ferment glucose, xylose, and arabinose, resolving the contradiction between ethanol production efficiency and sugar utilization versatility.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The modified yeast strain achieves multi-functionality by acquiring the ability to metabolize multiple sugar types (C6 sugars like glucose and C5 sugars like xylose and arabinose) through integrated gene expression. This universal sugar utilization capability allows a single strain to perform what previously required multiple specialized strains, directly addressing the adaptability limitation.

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

2Adaptability or versatility

If multiple genes are integrated into the cell genome, then sugar conversion capability is enhanced, but cell complexity increases

Engineering Contradiction:
Improvesugar conversion capabilityVSAvoidgenome complexity
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

Multiple gene functions are merged into the yeast genome through integrated expression cassettes. The xylA gene for xylose metabolism and the araA, araB, and araD genes for arabinose metabolism are combined with appropriate promoters and regulatory elements, creating a unified genetic system that enhances sugar conversion capability while managing genome complexity through organized integration.

Inventive Principle:
Principle #5Merging (Combining)

3Productivity

If glucose is the favoured carbon source, then growth is optimized, but galactose is not consumed under anaerobic conditions

Engineering Contradiction:
Improvegrowth rateVSAvoidcarbon source utilization
Core Design Contradiction:
ProductivityVSAdaptability or versatility

Solution Approach 1:

The patent modifies metabolic parameters by introducing heterologous genes that alter the yeast's carbon metabolism pathways. The integrated araA, araB, and araD genes enable arabinose metabolism, while xylA enables xylose metabolism, changing the parameter of carbon source utilization to include C5 sugars that were previously inaccessible, thereby resolving the preference issue without compromising growth optimization.

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

The mixed sugar cell effectively converts up to 90% of available glucose, xylose, arabinose, and galactose into fermentation products, enhancing the economic viability of renewable fuel production from lignocellulosic biomass.

Implementation Method 1

Heterologous expression of a xylose isomerase (XI) is an option for enabling yeast cells to metabolize and ferment xylose

Methodology Applied
Scientific EffectIsomerization:

Implementation Method 2

fermentation of the sugar composition in the presence of a mixed sugar cell

Methodology Applied
Scientific EffectFermentation: Fermentation

Data Source

PatentEP3241894A1Cell suitable for fermentation of a mixed sugar composition
Publication Date: 2017.11.08 DSM IP ASSETS BV
  • EP3241894A1 patent drawingFigure 1
  • EP3241894A1 patent drawingFigure 2
  • EP3241894A1 patent drawingFigure 3

AI summary

The present invention relates to a cell suitable for production of one or more fermentation product from a sugar composition comprising glucose, galactose, arabinose and xylose, wherein the cell comprises two to fifteen copies of one or more xylose isomerase gene or two to fifteen copies of one or more xylose reductase and xylitol dehydrogenase, and two to ten copies of araA, araB and araD, genes, wherein these genes are integrated into the cell genome.