Metabolically Engineered Cells for Stable Di- and Oligosaccharide Production
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Solution Overview
Problem
Metabolically engineered cells for di- and oligosaccharide production often suffer from clonal instability and transgene silencing due to the introduction of multiple coding DNA sequences, leading to inefficient production systems.
Innovation Solution
Genetically modified cells with multiple coding DNA sequences that differ in nucleotide sequence but encode polypeptides with the same function, allowing for efficient expression and overexpression of enzymes involved in di- and oligosaccharide production, such as glycosyltransferases and nucleotide-activated sugar synthesis enzymes, without clonal instability or transgene silencing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multiple coding DNA sequences are introduced into metabolically engineered cells to improve di- and oligosaccharide production, then production yield and productivity are enhanced, but clonal instability and transgene silencing occur
Solution Approach 1:
The patent changes the nucleotide sequence parameters of coding DNA sequences while maintaining the encoded polypeptide function. By using synonymous codons and silent mutations, the invention achieves different nucleotide sequences that encode identical or functionally equivalent polypeptides, thereby preventing clonal instability and transgene silencing while maintaining high productivity
Solution Approach 2:
The patent creates multiple copies of coding DNA sequences with identical or equivalent function but different nucleotide sequences. These copied sequences are introduced into the cell to provide functional redundancy and prevent silencing, ensuring stable and sustained high-level expression of enzymes needed for di- and oligosaccharide production
2Productivity
If multiple coding DNA sequences are introduced to enhance enzyme expression for di- and oligosaccharide synthesis, then production efficiency improves, but genetic heterogeneity increases
Solution Approach 1:
The patent modifies nucleotide sequence parameters without changing the encoded polypeptide amino acid sequence. By using synonymous codons and silent mutations, the invention creates genetic diversity at the DNA level while maintaining protein-level homogeneity, thus improving production efficiency without compromising genetic stability
Solution Approach 2:
The patent applies local changes to specific nucleotide sequences while maintaining the overall genetic architecture. By targeting only the nucleotide level for diversification while keeping polypeptide sequences identical or equivalent, the invention achieves localized genetic variation that enhances expression without creating harmful heterogeneity
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 approach results in improved yield, productivity, and growth speed of the cells, providing a more efficient method for producing di- and oligosaccharides compared to cells without these genetic modifications.
Implementation Method 1
one or more glycosyltransferases that are expressed and/or over-expressed by the cells and that catalyze the selective transfer of a sugar moiety from an activated nucleotide-sugar donor onto one or more saccharide acceptors
Data Source
AI summary
The disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, the disclosure is in the technical field of metabolically engineered cells and use of the cells in a cultivation or fermentation. The disclosure describes a cell and a method for production of a di- and/or oligosaccharide. The cell comprises a pathway for production of the di- and/or oligosaccharide and is genetically modified for expression and/or overexpression of at least one set of multiple coding DNA sequences wherein the multiple coding DNA sequences within one set differ in nucleotide sequence and each encode a polypeptide, wherein the polypeptides have the same function and/or activity of interest. Furthermore, the disclosure provides for purification of the di- and/or oligosaccharide from the cultivation.