5'-Ribonucleotide Production via pH-Controlled Cell Wall Separation
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Solution Overview
Problem
Existing processes for producing 5'-ribonucleotides result in low yields and are not suitable for applications requiring a clean taste due to high levels of amino acids and yeast components, leading to turbidity issues.
Innovation Solution
A process involving autolysis of microorganisms at specific pH conditions, followed by solid/liquid separation and conversion of the RNA-containing cell wall fraction into 5'-ribonucleotides using 5'-phosphodiesterase, with pH control between 1.0 and 5.1 to maximize 5'-ribonucleotide production and minimize turbidity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional autolysis process is used to produce 5'-ribonucleotides, then the production process is simple, but the yield of 5'-ribonucleotides is low and the taste is not clean due to high levels of amino acids and yeast components
Solution Approach 1:
The patent applies parameter changes by controlling the pH during solid/liquid separation to be between 2.0 and 4.5 (optimally 3.0-4.0), which is lower than conventional processes. This pH control causes RNA to remain associated with the cell wall fraction rather than dissolving into the supernatant, enabling selective recovery of RNA-containing cell walls and production of clean-tasting 5'-ribonucleotides without excessive amino acids and yeast components
Solution Approach 2:
The patent segments the autolysate into distinct fractions through pH-controlled solid/liquid separation. The RNA-containing cell wall fraction is separated from the supernatant containing amino acids and other yeast components. This segmentation allows selective processing of the cell wall fraction to produce 5'-ribonucleotides with clean taste, while the supernatant can be used separately or discarded
2Productivity
If partial extraction of cell content is performed to increase 5'-ribonucleotide content, then the 5'-ribonucleotide yield increases, but the turbidity of the yeast extract increases
Solution Approach 1:
The patent extracts only the necessary component (RNA) from the cell wall fraction through enzymatic treatment with nucleases and phosphodiesterases, while leaving the cell wall structure intact. This selective extraction produces clear 5'-ribonucleotide solutions without the turbidity associated with partial cell content extraction, because the cell wall matrix remains as a solid residue and does not contaminate the final product
3Productivity
If conventional solid/liquid separation is performed at pH 5.1 or higher, then the separation process is straightforward, but the RNA is not effectively recovered in the cell wall fraction leading to low 5'-ribonucleotide production
Solution Approach 1:
The patent changes the pH parameter from conventional values (5.1 or higher) to a lower range (2.0-4.5, optimally 3.0-4.0) during solid/liquid separation. This parameter change fundamentally alters the distribution of RNA, causing it to remain associated with the cell wall fraction rather than dissolving in the supernatant. The lower pH condition is maintained throughout the separation and washing steps to ensure maximum RNA recovery in the cell wall fraction
Solution Approach 2:
The patent applies preliminary action by adjusting the pH to the optimal range (3.0-4.0) before performing the solid/liquid separation. This preliminary pH adjustment ensures that RNA remains in the desired location (associated with cell walls) during the separation process, maximizing recovery efficiency before any subsequent processing steps
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 process achieves high yields of 5'-ribonucleotides, up to 99% w/w, and produces a clear yeast extract with low turbidity, suitable for applications requiring flavor enhancement without a bouillon-type taste.
Implementation Method 1
converting the RNA in the recovered RNA-containing cell wall fraction into 5'-ribonucleotides
Implementation Method 2
subjecting the autolysate to solid/liquid separation and recovering the RNA-containing cell wall fraction
Data Source
AI summary
The present invention describes a process comprising a) subjecting a microorganism to autolysis under conditions at which a substantial part of the RNA remains in a form degradable into 5-ribonucleotides; b) subjecting the autolysate to solid/liquid separation and recovering the RNA-containing cell wall fraction; and c) converting the RNA in the recovered RNA-containing cell wall fraction into 5- ribonucleotides, whereby the solid/liquid separation in step b) is done at a pH of lower than 5.1. Said process is simple and allows for the production of a very pure composition of 5'-ribonucleotides. It also allows for the production of a very clear yeast extract.