Biosensing System Cofactor Recycling via Dehydrogenase Enzymes
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Solution Overview
Problem
Biosensing systems that require cofactors such as NADH and NADPH often deplete these high-energy forms, leading to reduced functionality, as there is no efficient method to regenerate them within the system, limiting the duration and effectiveness of the biosensing process.
Innovation Solution
Incorporating a whole cell biocomponent with oxygenase and dehydrogenase enzymes, such as toluene ortho-monooxygenase and formate dehydrogenase, where the dehydrogenase enzyme regenerates NADH and NADPH using formate as a substrate, either delivered through a capillary or by diffusion, within a biosensing system that includes a transducer layer for detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If cofactors such as NADH and NADPH are used in biosensing systems, then the detection capability is improved, but the operational duration is reduced due to cofactor depletion
Solution Approach 1:
The patent implements a cofactor regeneration system where dehydrogenase enzymes (formate dehydrogenase, lactate dehydrogenase, or glucose dehydrogenase) continuously regenerate NADH and NADPH from their oxidized forms using substrate oxidation (formate, lactate, or glucose). This recovering mechanism converts depleted cofactors back to their active reduced forms, extending the biosensing system's operational duration without compromising detection capability
Solution Approach 2:
The patent establishes continuous cofactor regeneration through the integration of dehydrogenase enzymes that constantly convert oxidized cofactors (NAD+, NADP+) back to their reduced forms (NADH, NADPH) using ongoing substrate oxidation. This continuous regeneration ensures the biosensing system maintains its detection function indefinitely, transforming a finite resource system into a sustainable one
2Duration of action of moving object
If dehydrogenase enzymes are added to regenerate cofactors, then the operational duration is extended, but the system complexity increases
Solution Approach 1:
The patent merges the cofactor regeneration function directly into the biosensing system by integrating dehydrogenase enzymes into the same system architecture. The dehydrogenase enzymes are positioned in proximity to the oxygenase enzymes and cofactors, allowing in-situ regeneration without requiring separate external systems. This combining approach extends operational duration while minimizing the increase in system complexity
Solution Approach 2:
The patent implements self-service by enabling the biosensing system to regenerate its own cofactors through integrated dehydrogenase enzymes that use readily available substrates (formate, lactate, or glucose). The system becomes self-sufficient by internally converting depleted cofactors back to active forms, reducing the need for external intervention or complex external regeneration equipment
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 enables prolonged and consistent detection of analytes by maintaining the necessary cofactor levels, extending the operational life of the biosensing system and ensuring continuous, accurate measurements.
Implementation Method 1
a dehydrogenase enzyme that catalyzes the reaction of an oxidized cofactor selected from the group consisting of NAD+, NADP+, FAD, FADH, FMN, and FMNH and an electron donor
Implementation Method 2
the capillary tube is disposed to deliver the formate through the distal tip to the whole cell biocomponent
Implementation Method 3
the substrate is delivered through diffusion to the biocomponent
Data Source
AI summary
The present disclosure relates to biosensing systems and biosensing elements having increased storage capability and increased functional lifetimes through using compositions and methods for recycling cofactors.