Freeze-Dried Bacterial Cells Cryoprotection Stability
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Solution Overview
Problem
Bacterial cells used in food and pharmaceutical products are sensitive to production conditions and environmental factors, leading to reduced viability and stability, especially in hot and humid climates, making it challenging to maintain their efficacy over time.
Innovation Solution
A method involving the use of pyrophosphate ions, polyhydroxy substances, and L-cysteine in a cryoprotection solution to prepare freeze-dried bacterial cells with enhanced cell membrane integrity, resulting in a biomass with prolonged stability and viability, suitable for storage in zones IV.A and IV.B.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If bacterial cells are stored in hot and humid climates (zones IV.A and IV.B), then distribution and market access are improved, but cell viability and stability are reduced due to sensitivity to temperature and humidity
Solution Approach 1:
The patent introduces an oxygen scavenging system as an intermediary between the bacterial cells and the external environment. This system includes oxygen scavenging compounds (such as ascorbic acid, tocopherols, or enzymatic systems like glucose oxidase) that actively remove oxygen from the storage environment, creating a protective microenvironment that maintains cell viability while allowing storage in hot and humid conditions.
Solution Approach 2:
The patent creates an inert storage environment by removing oxygen through scavenging systems and replacing it with nitrogen or carbon dioxide atmospheres. This inert environment protects the bacterial cells from oxidative damage and maintains stability during storage in climatic zones IV.A and IV.B, enabling distribution to previously inaccessible markets.
2Reliability
If exposure to oxygen is prevented to maintain viability, then cell stability is improved, but the complexity of the preservation system increases due to need for oxygen scavengers and inert atmospheres
Solution Approach 1:
The patent implements self-service oxygen scavenging systems that automatically maintain the inert atmosphere without external intervention. Oxygen scavenging compounds are included in the formulation or packaging that continuously consume oxygen and maintain low oxygen levels throughout the shelf life, eliminating the need for complex active monitoring and control systems.
Solution Approach 2:
The patent simplifies the preservation system by changing the chemical parameters of the storage environment - using oxygen scavenging compounds that chemically bind oxygen and transform it into harmless products. This chemical approach is simpler than mechanical oxygen removal systems and maintains stability through parameter control rather than complex device management.
3Duration of action of stationary object
If freeze-drying is used to improve stability, then shelf-life is extended, but cell membrane integrity may be compromised during the drying process
Solution Approach 1:
The patent applies beforehand cushioning by incorporating protective agents in the formulation before freeze-drying. These agents include antioxidants (vitamin C, vitamin E), osmoprotectants (sugars, polyols), and membrane-stabilizing compounds that are present in the frozen state during drying, cushioning the cells against mechanical and oxidative stresses of the freeze-drying process.
Solution Approach 2:
The patent uses composite protective formulations that combine multiple protective agents - antioxidants, osmoprotectants, and membrane stabilizers - working synergistically during freeze-drying. This composite approach provides multi-layered protection that maintains membrane integrity while achieving long shelf-life through freeze-drying.
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
The method achieves a biomass of freeze-dried bacterial cells with prolonged shelf-life and increased viability, ensuring effective performance even under extreme storage conditions, as demonstrated by the stability and activity maintained over several months to years.
Implementation Method 1
A method involving the use of pyrophosphate ions, polyhydroxy substances, and L-cysteine in a cryoprotection solution to prepare freeze-dried bacterial cells with enhanced cell membrane integrity
Implementation Method 2
a biomass of freeze-dried, high-concentration and stable bacterial cells
Implementation Method 3
the method used to produce them
Data Source
AI summary
A biomass of freeze-dried bacterial cells and related devices, compositions and method of preparation are described. The method comprises (i) fermenting a previously prepared biomass of bacterial cells (bacterial biomass) to obtain a biomass of fermented bacterial cells (fermented biomass); (ii) concentrating the fermented biomass obtained from step (i) up to obtaining a biomass of concentrated bacterial cells (concentrated biomass) having a bacterial cell concentration comprised from 1×106 cells/ml of liquid biomass to 1×1012 cells/ml of liquid biomass; (iii) mixing the concentrated biomass obtained from step (ii) with a solution comprising or, alternatively, consisting of: (a) at least one phosphorous salt, and (b) at least one polyhydroxy substance to obtain a cryoprotected biomass of bacterial cells (cryoprotected biomass); (iv) freeze-drying the cryoprotected biomass obtained from step (iii) to obtain a biomass of freeze-dried bacterial cells (freeze-dried biomass).