Abstract: The invention relates to the technical field of bioengineering and provides a method for improving the specificity and affinity of an aptamer. The method includes: S1, screening a target of an aptamer from a compound information database by virtual computing; S2, verifying the screening result in Step S1 through experiments; S3, performing virtual saturation mutation on a site of the aptamer, and screening out a mutation site of the aptamer; S4, performing base substitution to the mutation site of the aptamer; and S5, detecting the binding parameter of the aptamer after base substitution with the target screened in Step S1, and selecting an aptamer with improved specificity and affinity after base substitution. An efficient molecular design-guided method is developed by computer rational calculation, to improve the specificity and binding affinity of the aptamer by directional modification. The present invention is of great significance for the practical application of aptamers.

Abstract: Described is a method for screening hydrolases with improved robustness by combining high-pressure molecular dynamics simulation and free energy calculation. The disclosed methods belongs to the fields of computational chemistry, bioinformatics, genetic engineering, and protein engineering, where the above method constructs and screens hydrolase mutants. The frequency of positive mutants in the screen is as high as 62.5%, meaning that the screening workload is effectively reduced, and the obtained mutants have high robustness and high industrial application potential.

Abstract: The invention relates to the technical field of bioengineering and provides a method for improving the specificity and affinity of an aptamer. The method includes: S1, screening a target of an aptamer from a compound information database by virtual computing; S2, verifying the screening result in Step S1 through experiments; S3, performing virtual saturation mutation on a site of the aptamer, and screening out a mutation site of the aptamer; S4, performing base substitution to the mutation site of the aptamer; and S5, detecting the binding parameter of the aptamer after base substitution with the target screened in Step S1, and selecting an aptamer with improved specificity and affinity after base substitution. An efficient molecular design-guided method is developed by computer rational calculation, to improve the specificity and binding affinity of the aptamer by directional modification. The present invention is of great significance for the practical application of aptamers.

Abstract: The disclosure discloses improvement of the robustness of hydrolases by combining high-pressure molecular dynamics simulation and free energy calculation, and belongs to the fields of computational chemistry, bioinformatics, genetic engineering and protein engineering. The method of the disclosure includes the following steps: first, searching for high robustness related regions and key amino acid sites by high-pressure disturbance combined with molecular dynamics model simulation calculation; then performing virtual saturation mutagenesis on amino acids in the key regions to construct a mutant library; then predicting the stability of the mutants using multi-scale free energy calculation software from protein dynamics, protein thermodynamics and prediction algorithms, and excluding false positive mutants as much as possible after multiple rounds of virtual selection. By using the above method to construct and screen hydrolase mutants, the frequency of positive mutants is as high as 62.