The high-throughput strain screening and optimization system is mainly used to screen and optimize microbial strains through efficient automated methods, especially in industrial microbial fermentation, drug production, environmental governance and other fields. The methods for screening bacterial strains are usually based on the following techniques:
1. Medium screening
Choosing the appropriate culture medium: Strain screening typically requires selecting different culture media to promote the growth of target microorganisms and the production of products. Different components of the culture medium can affect the metabolic products of the strain, so it is necessary to design specific culture media based on the target product or target function.
Optimization of culture medium: By adjusting the carbon source, nitrogen source, minerals, and pH value in the culture medium, the metabolic product production capacity of the strain can be optimized.
2. High throughput screening technology
High throughput screening technology (HTS) allows for parallel detection of the characteristics of a large number of bacterial strains in a relatively short period of time. Common high-throughput screening methods include:
Microplate method: Use 96 well, 384 well, or higher flux microplates to quickly screen the growth and product synthesis performance of multiple strains.
Liquid culture detection: Through an automated liquid processing system, the growth rate and metabolite concentration of bacterial strains are automatically measured to screen out the strains with the best performance.
Fluorescence probe method: using fluorescent markers or dyes to quickly screen excellent strains by detecting changes in bacterial growth or metabolic products.
Automated microscopy method: Combining image analysis technology, using automated microscope scanning and colony morphology analysis to screen the best strains from large-scale samples.
3. Genomic screening
Gene editing: Using gene editing tools such as CRISPR/Cas9 to selectively modify microbial genomes, in order to enhance the production capacity of target products or optimize specific metabolic pathways.
Whole genome scanning: Screening potential strains through genome sequencing and analysis, with a focus on those with advantageous metabolic or tolerance genes.
Transcriptome analysis: By analyzing the gene expression profile of strains through RNA seq, strains exhibiting high activity in the target metabolic pathway are screened.
4. Metabolite screening
Chromatographic analysis method: High performance liquid chromatography (HPLC), gas chromatography (GC), thin layer chromatography (TLC) and other techniques are used to detect the metabolites of bacterial strains under specific culture conditions, and to screen for strains with high ability to produce metabolites.
Mass spectrometry (MS): Combining mass spectrometry analysis, quantitative and qualitative analysis of metabolites in bacterial strains is performed to further screen for strains with higher yields.
5. Resistance screening
Drug resistance testing: For specific resistance screening, such as antibiotic resistance or heavy metal tolerance, high-throughput screening can be performed through resistance plates or liquid culture to find strains that are adapted to specific environmental conditions.
Environmental adaptability screening: In some industrial fermentation processes, strains may need to work under specific environmental conditions, such as high temperature, low pH, or high salt environments. By screening under these extreme conditions, adaptive strains can be found.
6. Enzyme activity screening
Enzyme activity detection: If the target strain produces a certain enzyme, appropriate substrates can be used to detect enzyme activity, such as using pigment substrates to determine the catalytic reaction of the enzyme. This method can quickly screen strains with high enzyme activity.
Fluorescent enzyme-linked immunosorbent assay: Using fluorescent labeled substrates and detection systems, enzyme activity is detected through fluorescence changes, and strains with excellent performance are screened.
7. Comprehensive screening
Multi index screening: By combining multiple factors such as growth rate, product yield, tolerance, and metabolic characteristics of the strain, a comprehensive evaluation is conducted to select the optimal strain. This method is often combined with high-throughput automation platforms for operation, which can screen on a large scale and efficiently.
8. Application of automation platform
High throughput screening systems are typically equipped with automated platforms that can automate the processes of sample processing, cultivation, detection, and analysis. For example, by combining liquid processing robots and automated microscopes, it is possible to screen a large number of bacterial strains in a short period of time.
9. Data analysis and optimization
Big data analysis: Through statistical analysis of high-throughput screening data and machine learning analysis, further optimize screening strategies to improve the accuracy and efficiency of strain screening.
Machine learning algorithms: Use machine learning algorithms to analyze the effects of different parameters (such as temperature, pH, nutrient composition, etc.) during the cultivation process on the performance of bacterial strains, and automatically optimize screening strategies.
Through these methods, the high-throughput strain screening optimization system can quickly and effectively screen potential strains for subsequent research or industrial applications.