Cryoprotectant Composition for Freeze-Dried Lactic Acid Bacteria
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for freeze-drying lactic acid bacteria fail to achieve long-term stability and viability, particularly at room temperature, due to the limitations of existing cryoprotectants, which are often animal-derived and pose risks, and do not ensure sufficient survival rates for pharmaceutical applications.
Innovation Solution
A novel combination of cryoprotectants, including a starch hydrolysate, a glutamic acid salt, and a polyol, such as dextrane, sodium glutamate, and sorbitol, is used to enhance the viability and stability of freeze-dried lactic acid bacteria, eliminating the need for animal-derived compounds and improving texture for easy grinding and long-term storage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional cryoprotectants are used for freeze-drying lactic acid bacteria, then the process is simple, but the long-term stability and viability of bacteria are insufficient
Solution Approach 1:
The patent uses a composite cryoprotectant system consisting of multiple components: a polyol (sorbitol, mannitol, or xylitol), a starch hydrolysate (dextrin or maltodextrin), and optionally a protein source (skim milk powder or casein). This composite approach provides synergistic protection for bacterial viability during freeze-drying and long-term storage, resolving the contradiction between simplicity and effectiveness.
2Reliability
If animal-derived cryoprotectants are used, then the protective effect is sufficient, but safety risks increase for pharmaceutical applications
Solution Approach 1:
The patent replaces animal-derived cryoprotectants with plant-based or synthetic alternatives: polyols (sorbitol, mannitol, xylitol), starch hydrolysates (dextrin, maltodextrin), and plant-based proteins. These substitutes provide comparable protective effects while eliminating safety risks associated with animal-derived products, making the formulation suitable for pharmaceutical applications.
3Reliability
If existing cryoprotectant methods are used, then the process is straightforward, but the survival rates of bacteria are insufficient for pharmaceutical applications
Solution Approach 1:
The patent optimizes the concentrations of cryoprotectant components to achieve maximum bacterial survival. The formulation includes a polyol at 5-30% w/v, a starch hydrolysate at 5-30% w/v, and optionally protein at 1-10% w/v. These parameter optimizations significantly improve survival rates while maintaining ease of manufacture through straightforward mixing and freeze-drying processes.
4Ease of operation
If freeze-drying is performed without optimized cryoprotectants, then the process is simple and fast, but the texture of the lyophilized cake is poor for grinding
Solution Approach 1:
The starch hydrolysate (dextrin or maltodextrin) acts as an intermediary substance that modifies the matrix structure of the lyophilized cake. This component improves the physical texture and grindability of the dried product while the polyol provides cryoprotection. The combination achieves both fast processing and excellent texture properties suitable for pharmaceutical formulation.
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 novel cryoprotectant combination achieves high viable cell yields and maintains long-term stability, with over 50% retention of viability after 6 months, and up to 90% after 1 year, even at 25°C, significantly surpassing traditional methods in preserving bacterial viability and stability.
Implementation Method 1
addition of selected cryoprotectants to the biomass and subsequent lyophilization are crucial steps
Implementation Method 2
lyophilized (freeze-dried) forms
Implementation Method 3
improving the long term stability of the freeze dried bacteria at different temperature conditions
Data Source
AI summary
The present invention comprises the discovery and development of an effective cryoprotectant composition, without containing skim milk or any other animal-derived compounds, to achieve long-term stability of freeze-dried lactic acid bacteria (LAB), at different temperatures, whereby the retention of viability of the freeze-dried LAB after 6 months of storage, preferably after 9 months of storage, more preferably after 12 months of storage is more than 50%. The invention is in the field of producing freeze dried bacteria, in particular Lactic acid bacteria. More in particular, the invention relates to the use of a novel combination of cryoprotectants for increasing the viability of bacteria after freeze drying, improving the texture of the lyofilized cake for easy grinding and improving the long term stability of the freeze dried bacteria at different temperature conditions. The invention further relates to such freeze dried bacteria for use in food industry or in human or animal health applications. More in particular, the invention relates to the increased viability and long-term storage of recombinant bacteria capable of expressing heterologous proteins or peptides and administered to humans or animals for therapeutic or vaccination purposes.