Beta-Glucan Production via 1,3-Beta-D-Glucan Synthase Overexpression
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Solution Overview
Problem
Current methods for improving β-glucan production in microorganisms, such as Schizophyllum commune, are undirected and lack predictability, leading to inefficient and costly strain selection, as they do not allow targeted modification of β-glucan producing microorganisms.
Innovation Solution
Overexpressing 1,3-β-D-glucan synthase in microorganisms like Schizophyllum commune leads to significant increases in β-glucan production, utilizing strong promoters and introducing additional copies of the 1,3-β-D-glucan synthase gene to enhance expression levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If undirected methods (protoplast generation, crossing, random mutagenesis) are used to improve β-glucan production, then strain selection can be performed, but the process becomes inefficient and costly due to lack of predictability
Solution Approach 1:
The patent applies parameter changes by modifying the genetic parameters of the microorganism - specifically introducing additional copies of the 1,3-β-D-glucan synthase gene and using strong promoters to overexpress the enzyme. This directed genetic modification changes the biochemical parameters of β-glucan synthesis, leading to predictable increases in production yield without requiring time-consuming random strain selection processes
Solution Approach 2:
The patent employs preliminary action by pre-selecting and characterizing the 1,3-β-D-glucan synthase gene before introducing it into the microorganism. The gene is optimized with strong promoters and regulatory elements in advance, so that when introduced, it immediately provides the desired overexpression effect. This eliminates the need for subsequent screening and selection of multiple random mutants
2Productivity
If undirected methods are used to improve β-glucan production, then strain variability can be explored, but manufacturing precision and predictability deteriorate
Solution Approach 1:
The patent achieves manufacturing precision by making specific, controlled changes to genetic parameters - introducing defined numbers of gene copies with characterized promoter sequences. This directed approach ensures that the same genetic modification produces consistent results across different production batches, eliminating the variability inherent in random mutagenesis and crossing methods
Solution Approach 2:
The patent uses copying by introducing additional copies of the 1,3-β-D-glucan synthase gene into the microorganism genome. This creates multiple identical copies of the functional gene, ensuring consistent enzyme production across all cells. The copied gene sequences are identical and controlled, providing predictable and precise production outcomes rather than the random variations produced by traditional breeding methods
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 results in increased yields of β-glucans like schizophyllan and scleroglucan, with genetically modified microorganisms producing up to 2.2 times more polymer compared to non-modified controls, offering a targeted and efficient method for enhancing β-glucan synthesis.
Implementation Method 1
β-glucans comprise non-cellulosic polymers of β-glucose linked via glycosidic β(1-3) bonds
Implementation Method 2
polypeptide having 1,3-β-D-glucan synthase-activity
Data Source
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AI summary
The present invention relates to genetically modified microorganisms capable of producing beta-glucans, characterized in that said genetically modified microorganism overexpresses (i) a polynucleotide encoding a polypeptide having 1,3-?-D-glucan synthase-activity, and/or (ii) a polypeptide having 1,3-?-glucan synthase-activity, compared to a corresponding non-modified control microorganism of the same strain. The present invention also relates to the use of a polynucleotide encoding a polypeptide having 1,3-?-D-glucan synthase-activity or the use of such a polypeptide for producing ?-glucans. Furthermore, the present invention relates to methods for producing ?-glucans comprising the introduction of a promoter upstream of a polynucleotide encoding a polypeptide having 1,3-?-D-glucan synthase-activity thereby increasing the expression of said polynucleotide, or a polynucleotide encoding a polypeptide having 1,3-?-D-glucan synthase-activity into a microorganism being able to synthesize ?-glucans.