In the diagnosis of diabetic complications and the monitoring of metabolic diseases, the accuracy of blood ketone testing directly influences clinical decision-making. This precision relies fundamentally on two critical enzyme reagents: β-Hydroxybutyrate Dehydrogenase (HBDH) and Diaphorase (DPH). As indispensable components of the blood ketone detection system, these enzymes ensure measurement accuracy and stability, serving as the core drivers for the advancement of Continuous Ketone Monitoring (CKM) technology.

Industry Landscape: Small Market, Vital Impact

The current blood ketone monitoring sector is characterized by a "niche but essential" status. While it represents a smaller portion of many companies' diagnostic product lines compared to glucose monitoring, it is an irreplaceable link in metabolic disease management. Historically, the market for these core raw materials has been limited, with high-quality enzyme powders long dependent on Japanese imports. This has exposed the industry to supply chain risks, including potential shortages and price fluctuations driven by international trade tensions. Consequently, there is an urgent demand for high-quality, domestically produced enzyme reagents to achieve import substitution.

The "Relay Race" of Electrochemical Detection

The detection process of electrochemical blood ketone test strips functions like a molecular "relay race," involving a series of enzymatic reactions and electrochemical signal conversions. It is primarily completed in two steps:

β-Hydroxybutyrate Dehydrogenase (HBDH) – The "Starter": 

It catalyzes the conversion of β-hydroxybutyrate in the blood into acetoacetate. Simultaneously, it generates a reduced substance (NADH), achieving the initial conversion from a metabolite to a detectable chemical signal.

Diaphorase (DPH) – The "Efficient Messenger": 

It transfers electrons from the reduced substance to an oxidized electron mediator (such as Ru³⁺), converting it into a reduced form (such as Ru²⁺). This effectively transforms the biochemical reaction into a measurable electrical signal.

In short, these two enzymes work in seamless coordination—one "initiates the signal" and the other "transmits the signal"—to transform the invisible concentration of β-hydroxybutyrate into clear, quantifiable data.

β-Hydroxybutyrate Dehydrogenase (HBDH): The Precision Engine

As the "first gateway" of blood ketone detection, the performance of HBDH directly determines the foundational accuracy of the test. Hzymes has introduced an HBDH variant that delivers reliable performance and outstanding results across all metrics:

High Specific Activity: 

High enzyme protein activity effectively reduces reagent dosage while enhancing reaction efficiency.

Hzymes’ three sample batches demonstrate superior specific protein activity.

Superior Stability: 

After incubation at 37°C for 7 days, the enzyme retains over 95% of its activity, making it ideal for long-distance transport and long-term storage.

Enzyme activity remained above 90% across three batches after 7 days of incubation at 37°C.

Batch-to-Batch Consistency: 

The variance in enzyme and protein activity between different batches is less than 2%, ensuring the reproducibility of experiments and large-scale production.

The batch-to-batch coefficient of variation (CV) for both enzyme activity and protein activity is <2% across three batches.

Diaphorase (DPH): The Efficient Bridge for Signal Transmission

Diaphorase serves as the critical bridge for signal delivery; its transmission efficiency and stability directly impact the quality of the final detection signal. Hzymes’ Diaphorase offers distinct advantages:

Wide Stability Range: 

Maintains stable activity under 50°C (pH 7.5, 30 min at 1mg/mL) and remains stable within a pH range of 4.5–9.0 (25°C, 16 h at 1mg/mL), making it adaptable to diverse application scenarios.

The enzyme maintains stable activity below 50°C (Conditions: pH 7.5, 30 min, enzyme concentration at 1 mg/mL).

Enzyme activity remains stable within a pH range of 4.5–9.0 (Conditions: 25°C, 16 h, enzyme concentration at 1 mg/mL).

Excellent Thermal Stability & Consistency: 

High thermal stability and batch-to-batch consistency significantly reduce the complexity of reagent development.

Future-Ready: Powering Continuous Ketone Monitoring (CKM)

Continuous Ketone Monitoring (CKM) enables 24-hour real-time tracking, providing vital data for Diabetic Ketoacidosis (DKA) alerts and ketogenic diet management. This technology demands even higher long-term stability and sensitivity from enzymes.

The high activity and stability of HBDH allow for a sensitive response to low concentrations of β-hydroxybutyrate, even during prolonged operation. Meanwhile, the thermal resistance and broad adaptability of Diaphorase ensure consistent signal transmission despite fluctuations in internal or external environments. Currently, both enzymes are being integrated into the development of CKM sensors, significantly improving the reliability and practicality of next-generation monitoring technology.