ALDC Enzyme Stabilization via Zinc Supplementation
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Solution Overview
Problem
Diacetyl, an unwanted by-product of fermentation, negatively affects the flavor and taste of beers due to its strong smell, and existing enzymes like ALDC are unstable, especially in fermenting conditions with low malt content, limiting their effectiveness.
Innovation Solution
The use of ALDC enzymes in combination with zinc at specific concentrations, ranging from 1 μM to 200 mM, to enhance stability and activity, and the inclusion of additional enzymes like acetolactate reductoisomerases, improves the conversion of diacetyl to acetoin, reducing its negative impact.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If ALDC enzyme is used to convert diacetyl to acetoin, then diacetyl levels are reduced and flavor is improved, but the enzyme becomes unstable under fermenting conditions with low malt content
Solution Approach 1:
The patent applies parameter changes by modifying the chemical environment through zinc supplementation. Zinc ions alter the stability parameters of ALDC enzyme, enabling it to maintain activity under fermenting conditions with low malt content where it would otherwise become unstable. This resolves the contradiction by changing the chemical parameters of the system.
Solution Approach 2:
Zinc acts as an intermediary substance that mediates between the ALDC enzyme and the low malt content fermentation environment. The zinc ions bind to the enzyme and protect it from degradation, allowing the enzyme to function effectively in conditions that would normally cause instability. This intermediary resolves the adaptability issue without requiring modification of the enzyme itself or the fermentation substrate.
2Productivity
If ALDC enzyme activity is increased to reduce diacetyl formation, then flavor quality improves, but enzyme stability under fermenting conditions deteriorates
Solution Approach 1:
The patent uses parameter changes by introducing zinc ions to modify the stability parameters of the ALDC enzyme. This allows the enzyme to maintain high activity levels while becoming stable under fermenting conditions. The zinc supplementation changes the chemical parameters of the system, enabling simultaneous achievement of high productivity and stability.
Solution Approach 2:
The solution creates a composite system consisting of ALDC enzyme and zinc ions. This composite maintains the catalytic activity of the enzyme while the zinc component provides stability under fermentation conditions. The composite material approach allows both high enzyme activity and stability to coexist, resolving the contradiction between productivity and stability.
3Reliability
If zinc is added to stabilize ALDC enzyme, then enzyme stability and activity improve, but the complexity of the fermentation process increases
Solution Approach 1:
The patent applies parameter changes by adding zinc ions, which modifies the chemical environment of the fermentation process. While this does increase process complexity slightly, the change is minimal and involves simply supplementing the fermentation medium with zinc. The benefit of improved enzyme stability and activity outweighs the minor increase in process complexity.
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 stabilizes and increases the activity of ALDC enzymes, effectively reducing diacetyl levels in fermented beverages, thereby improving the flavor and aroma of beers by converting diacetyl into the more acceptable acetoin.
Implementation Method 1
A composition comprising an acetolactate decarboxylase (ALDC) enzyme and zinc, where the zinc is present at a concentration of about 1 μM to about 200 mM
Data Source
AI summary
The present disclosure provides methods, compositions, apparatuses and kits comprising ALDC enzymes having a better stability and/or activity, and, optionally, the yield of ALDC enzymes which can be recovered from microorganisms is improved. In some embodiments, the present disclosure provides methods, apparatuses, compositions and kits for the use of metal ions to increase stability and/or activity, and which further can be used to recover the enzymes from microorganisms in improved yields.