(p)ppGpp Synthetase Variant for Higher 5'-Inosinic Acid Yield
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Solution Overview
Problem
Existing methods for producing 5'-inosinic acid using microorganisms are limited by the need for extensive research on protein activity changes to enhance productivity, particularly in enzymes, transcription factors, and transport proteins involved in its biosynthetic pathway.
Innovation Solution
A (p)ppGpp synthetase variant with an amino acid substitution at position 746 from glycine to aspartic acid, encoded by a specific nucleotide sequence, is introduced into a Corynebacterium strain to enhance 5'-inosinic acid production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If extensive research on protein activity changes is conducted to enhance productivity, then 5'-inosinic acid production efficiency can be improved, but the complexity and time required for research and development increases
Solution Approach 1:
The patent applies parameter changes by modifying a specific amino acid parameter in the (p)ppGpp synthetase protein sequence. Specifically, glycine at position 746 is substituted with aspartic acid, which changes the protein's catalytic properties and increases 5'-inosinic acid production efficiency without requiring extensive research on multiple protein activities
Solution Approach 2:
The patent segments the complex biosynthetic pathway by focusing on a specific enzyme ((p)ppGpp synthetase) and a specific residue (position 746) within that enzyme. This segmentation allows targeted modification rather than comprehensive analysis of all involved proteins, reducing development complexity while maintaining productivity improvement
2Productivity
If multiple genes and proteins involved in biosynthetic pathway are targeted for modification, then 5'-inosinic acid productivity can be increased, but the time and resources required for research increases
Solution Approach 1:
The patent applies preliminary action by pre-identifying and pre-modifying the critical amino acid residue at position 746 in the (p)ppGpp synthetase enzyme before implementing the production process. This preliminary identification of the key functional residue eliminates the need for extensive systematic screening of multiple genes and proteins, significantly reducing research time while achieving productivity enhancement
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 variant strain exhibits increased 5'-inosinic acid productivity, producing up to 50% more than the parent strain, demonstrating improved efficiency in biosynthesis.
Implementation Method 1
a (p)ppGpp synthetase variant consisting of the amino acid sequence of SEQ ID NO: 2 in which glycine at position 746 in the amino acid sequence of SEQ ID NO: 4 is substituted with aspartic acid
Data Source
Figure 1
AI summary
The present invention relates to a novel (p)ppGpp synthetase variant and a method of producing 5'-inosinic acid using the same. The (p)ppGpp synthetase variant is obtained by substituting one or more amino acids in the amino acid sequence constituting (p)ppGpp synthetase to change the activity of the protein, and a recombinant microorganism comprising the (p)ppGpp synthetase variant is capable of efficiently producing 5'-inosinic acid.