Enzymatic Biosensor for Onsite Yeast Nitrogen Detection
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Solution Overview
Problem
Current methods for measuring yeast assimilable nitrogen (YAN) levels in wine grapes are slow, expensive, and require complex processes, making them unsuitable for routine onsite testing, particularly in the wine fermentation process.
Innovation Solution
Development of two- and three-enzyme systems, including BpsA (blue pigment synthetase A), GS (glutamine synthetase), and ArgZ (arginine dihydrolase), which convert specific nitrogen-containing molecules into indigoidine, allowing for quantitative or qualitative assessment of YAN levels using colorimetric assays.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If HPLC analysis is used to measure FAN levels, then measurement precision is improved, but device complexity and time consumption increase
Solution Approach 1:
The patent extracts the specific measurement function for FAN from the complex HPLC system and implements it through a dedicated enzymatic assay using glutamine synthetase and blue pigment synthetase A. This specialized enzyme-based approach isolates the measurement capability needed for routine testing, eliminating the need for the entire HPLC system while maintaining measurement accuracy for the specific analyte of interest.
Solution Approach 2:
The patent employs disposable test strips or single-use reagent systems containing the enzyme cocktails, replacing the expensive and complex HPLC equipment. These simple, inexpensive test systems are designed for single-use or limited-use scenarios, making them economically viable for routine onsite testing while eliminating the need for costly laboratory instrumentation.
2Measurement precision
If enzymatic test kits requiring UV/vis spectrophotometer are used, then measurement precision is improved, but ease of operation deteriorates due to equipment requirements
Solution Approach 1:
The patent extracts the colorimetric detection function from the UV/vis spectrophotometer and implements it through visual color comparison or simple photodetection on test strips. The enzymatic reactions produce colored products (indigoidine pigments) that can be quantified by simple optical methods, eliminating the need for expensive spectrophotometric equipment while maintaining sufficient measurement precision for routine applications.
Solution Approach 2:
The patent replaces the mechanical/optical complexity of UV/vis spectrophotometry with simpler detection methods. The enzymatic colorimetric reactions produce visible color changes that can be measured by basic photodetectors, smartphone cameras, or even visual comparison against reference standards, substituting complex instrumentation with simpler alternatives that are suitable for onsite use.
3Measurement precision
If current commercial laboratory testing methods are used, then measurement precision is improved, but productivity deteriorates due to slow turnaround time
Solution Approach 1:
The patent employs pre-prepared enzyme cocktails and reagent systems that are ready for immediate use, eliminating the need for complex sample preparation and instrument calibration required by HPLC and spectrophotometric methods. The enzymatic reactions are designed to proceed rapidly under optimized conditions, providing quick results without compromising measurement accuracy, thus enabling fast turnaround for routine monitoring applications.
Solution Approach 2:
The patent optimizes reaction conditions including pH, temperature, and enzyme concentrations to maximize reaction speed while maintaining measurement precision. By adjusting these parameters, the enzymatic assays achieve rapid completion times suitable for high-throughput routine testing, contrasting with the slow, multi-step procedures of traditional laboratory 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
Enables rapid, cost-effective, and onsite detection of key nitrogen-containing molecules, improving the monitoring of fermentation processes and wine quality by providing accurate YAN levels without the need for expensive equipment or laboratory testing.
Implementation Method 1
blue pigment synthetase A (BpsA) enzyme... capable of converting glutamine to indigoidine pigment
Implementation Method 2
glutamine synthetase (GS) enzyme... capable of converting glutamate and ammonium into glutamine
Implementation Method 3
arginine dihydrolase (ArgZ) enzyme... capable of converting arginine to ornithine and ammonium
Implementation Method 4
The BpsA enzyme... converts the glutamine to the blue pigment indigoidine... analysing the indigoidine pigment produced
Data Source
AI summary
Biosensor systems for detecting levels of nitrogen-containing molecules are provided. Further provided are biosensor system components and various detection platforms, methods, and kits.