Protease circuits for processing biological information.

Area of Science:

• Synthetic Biology
• Biochemistry
• Molecular Engineering

Background:

• Engineered biocircuits augment living functions using biological components.
• Proteases are enzymes that can be leveraged for biological information processing.

Purpose of the Study:

• To demonstrate the integration of proteases into digital and analog biocircuits.
• To develop cell-free systems for quantifying bacterial burden and calculating selective drug doses.
• To utilize protease activity for solving complex mathematical problems like Learning Parity with Noise (LPN).

Main Methods:

• Constructed peptide-caged liposomes to represent protease activity as binary (0 or 1) signals for Boolean logic operations.
• Assembled a cell-free biocircuit capable of interacting with bacteria-containing blood.
• Developed multi-valued (analog) signal processing by controlling enzyme sharing between substrates and manipulating substrate concentrations.

Main Results:

• Successfully created biological equivalents of Boolean logic gates, comparators, and analog-to-digital converters using protease activity.
• Demonstrated a cell-free biocircuit that quantifies bacterial burden and calculates a selective drug dose.
• Showcased the ability of protease-based analog signals to solve the Learning Parity with Noise (LPN) problem.

Conclusions:

• Protease activity can be effectively utilized for processing biological information through both binary Boolean logic and multi-valued analog signals.
• This work expands the capabilities of engineered biocircuits and offers new tools for diagnostics and computation.