17 results about "Rational design" patented technology

This invention belongs to the field of enzyme engineering technology, specifically providing a nitrile hydratase mutant and its application. The nitrile hydratase mutant is derived from *Aurantimonas manganoxydans*. Am NHase)SI859A was modified using rational design methods based on its three-dimensional protein structure. The resulting nitrile hydratase mutant ultimately achieved an enzyme activity that was up to 6.74 times higher than that of the wild enzyme, and its stability did not decrease significantly within two hours under heating conditions at 50°C.

The invention belongs to the technical field of molecular biology, and particularly discloses a theophylline aptamer and a virtual screening method and application thereof. The theophylline aptamer is selected from one or more of MUT9, MUT16, MUT67, MUT78 or MUT89. The invention further discloses a preparation method of the theophylline aptamer. The invention discloses a theophylline aptamer as well as a virtual screening method and application thereof, the theophylline aptamer is high in affinity and specificity, the virtual screening method remarkably improves the accuracy and efficiency of virtual screening, the theophylline aptamer can be quickly obtained at low cost, and a powerful calculation tool is provided for rational design and application of the aptamer.

The application discloses a dye-decolorizing peroxidase mutant and application thereof, and a novel dye-decolorizing peroxidase GA1DyPB screened from Rhodococcus globerulus GA1 is used as a starting enzyme, computer-aided semi-rational design enzyme engineering is used for site-directed modification and saturation mutation modification, and comparative analysis is conducted on the enzymatic properties of the mutant and the original enzyme, so as to provide a theoretical basis and technical support for subsequent directional mutation and catalytic performance improvement of the dye-decolorizing peroxidase. Finally, lignin is used as a substrate to conduct long-time lignin enzymatic depolymerization reaction for 48 hours, and the ability of the mutant in the lignin depolymerization is verified.

This application relates to the technical fields of synthetic biology and enzyme engineering, and in particular to a computational-experimental closed-loop rational design method for polyketide synthases. The method includes: constructing a full-length FCHS model using AlphaFold3; identifying binding hotspots and verifying dynamic stability through molecular docking and kinetic simulations; and constructing a low-redundancy mutant library containing A, L, and F substitutions. This application solves the problems of difficult protein structure analysis and limited experimental throughput through a closed-loop process of computational pre-screening, dynamic verification, and evolutionary filtering, providing an efficient paradigm for the improvement of complex multifunctional enzymes.

This invention discloses a PLA-degrading lipase mutant and its application. First, the lipase PaPlaA gene with the amino acid sequence shown in SEQ ID NO.1 was selected. Highly efficient heterologous expression was achieved by constructing a PgapA-T7 dual promoter system and optimizing the RBS sequence through ribosome binding site design, resulting in a recombinant enzyme activity yield of 6.25 U / mL, approximately 2.5 times higher than the original strain. Then, combining molecular dynamics simulations and rational design, the substrate pocket and flexible region were structurally modified to obtain the PLA-degrading lipase mutant. The obtained mutant exhibited excellent catalytic performance when applied to PLA degradation. This invention also provides a composite enzyme preparation obtained by combining the PLA-degrading lipase mutant with the protease nprT612. This composite enzyme preparation can achieve a degradation rate of up to 78.2% for PLA nonwoven fabrics. This provides a stable enzyme catalyst and a feasible synergistic degradation process for the efficient biodegradation of PLA at high temperatures.

This invention discloses a biomimetic dual-scale lattice metamaterial with high damage tolerance and electromagnetic absorption performance, and its design method. It belongs to the field of integrated design technology of damage tolerance design and electromagnetic function. This invention uses finite element simulation software, based on a first-order unit lattice, and adopts a recursive modeling method to design a second-order dual-scale lattice structure. By rationally designing the topology and size parameters of the dual-scale lattice, the structure maintains a high electromagnetic absorption rate over a wide frequency range, improves the damage tolerance of the structure, and, while maintaining a low relative density, makes it have high specific stiffness and specific strength, thus achieving mechanical-electromagnetic integrated optimization.

The application discloses a chondroitin-4-O-sulfotransferase mutant construction method and belongs to the field of bioengineering technology. The application is characterized in that a wild-type chondroitin-4-O-sulfotransferase (C4ST) is precisely reformed through a calculation-guided semi-rational design, a SUMO label and an AE connecting peptide are fused at an N end to improve soluble expression, and on this basis, a three-point combined mutant V74L / M117T / Y235V is constructed through a site-directed mutagenesis technology, the enzyme activity of the mutant in a 5L fermenter is as high as 76328 U / L, the specific enzyme activity is 327 U / mg, and the enzyme activity and the specific enzyme activity are 16.6 times and 1.3 times of the wild type, respectively. The mutant is used for catalyzing preparation of chondroitin sulfate A, the sulfation degree can reach 97.0% after 48 h of reaction at 37 DEG C. The application significantly improves the catalytic efficiency of C4ST and provides a key technology of efficient and stable enzyme method for preparing CSA.

This invention relates to an endonuclease β-agarase, its mutants, mutation methods, and applications, belonging to the field of biotechnology. This invention discloses for the first time a mutant of *Pseudomonas alterniflora* (…). Pseudoalteromonas An endonuclease β-agarase, ORF2093, was obtained from strain A601. This endonuclease β-agarase ORF2093 exhibits certain thermal stability, wide pH tolerance, and stable physicochemical properties, thus possessing potential for industrial application. This invention employs rational design and site-directed mutagenesis technology to mutate the serine residue at position 373 in the non-catalytic region of the endonuclease β-agarase ORF2093 to asparagine. The resulting mutant showed significantly enhanced relative activity compared to the wild type, and it was found that the mutant enzyme possesses the ability to degrade novel agaroctaose substrates.

This invention discloses a cephalothinyl synthase mutant and its application in the biosynthesis of cephalothinyl synthase, belonging to the fields of enzyme engineering and synthetic biology. Through rational design and mutation screening of tobacco-derived cephalothinyl synthase (NtCBTS), this invention obtained a mutant with significantly improved catalytic performance. Specifically, the cephalothinyl synthase yield of strain containing the mutant NtCBTS_K208R-A268C-K353R was increased by 231.18% compared to strains containing NtCBTS. Applying this mutant to a *Saccharomyces cerevisiae* expression system achieved highly efficient biosynthesis of cephalothinyl synthase, significantly improving the production level of the target product. The highly active enzyme mutant and its application strategy provided by this invention effectively improve the biosynthetic efficiency of cephalothinyl synthase, providing important technical support for constructing high-yield yeast cell factories and promoting its industrial production.

The application discloses a high-activity acetaldehyde dehydrogenase mutant, a preparation method and application thereof. The mutant is obtained by rational design on acetaldehyde dehydrogenase ALDHTt from thermophilic thermus, and is specifically that an asparagine at the 159th position is mutated into tyrosine, and is recorded as N159Y. Enzymatic property characterization shows that the specific enzyme activity of the mutant N159Y is 2.08 times of that of the wild type, the catalytic efficiency is 5.3 times of that of the wild type, and is 2.96 times of that of human ALDH2. Molecular dynamics simulation reveals that the flexibility of the mutant is moderately enhanced in the vicinity of the substrate binding channel and the cofactor binding site, which is beneficial to reduce the substrate entry energy barrier and promote the cofactor binding and release, and meanwhile, the overall structural stability is not adversely affected. The mutant overcomes the defects of human ALDH2, such as difficult expression and poor stability, and can be used as a core enzyme element for constructing an alcohol metabolism multi-enzyme cascade system or an oral enzyme preparation, and has a wide application prospect in preparation of products for alcohol metabolism or acetaldehyde clearance.

The application discloses a method for analyzing gibberellin GA4 high-yield mechanism of Gibberella fujikuroi based on comparative transcriptomics, and belongs to the field of industrial microorganisms. The method constructs a correlation model of "specific genotype strain-standardized fermentation-time sequence transcriptome analysis", and systematically analyzes the dynamic regulation network of product synthesis through high-fidelity sampling at multiple time points, high-throughput sequencing, differential expression analysis, time sequence clustering and weighted gene co-expression network analysis. The core lies in converging massive data into a short list of key target genes stably associated with high-yield traits, including rate-limiting enzymes, transport proteins and regulatory factors, so as to guide the rational design of metabolic engineering and strain improvement. The application forms a standardized and iterative systematic solution, and provides clear molecular targets and efficient methodological tools for breaking through the yield bottleneck of natural product fermentation.

The application belongs to the field of enzymatic engineering and computer-aided biological technology, and particularly relates to an endochitinase mutant, a coding gene thereof and application, and an amino acid sequence of the endochitinase mutant is shown as SEQ ID NO. 4 or SEQ ID NO. 6. In order to solve the problems of low catalytic efficiency and poor stability of natural endochitinase, the application obtains N197R and N197RCBM14 fusion mutants by computer-aided rational design on wild type ThChi8. Experiments prove that the specific activity of the key mutant N197RCBM14 is increased to 268.25% of the wild type. The mutant gene can be cloned into a pET-29a(+) vector and expressed in Escherichia coli Rosetta (DE3), and the secreted enzyme preparation can efficiently degrade chitin to produce chitooligosaccharides, and has a wide application prospect in the fields of biomass conversion, agriculture and medicine.

The application discloses a cephalin cytidylyltransferase mining method based on a deep learning model. The method breaks through the limitation of traditional homologous comparison, and can identify CCT candidate enzymes with high catalytic activity, excellent solubility and suitable for engineering application in a protein sequence space of hundreds of millions of levels with higher screening efficiency and prediction accuracy. On this basis, rational design and zero-sample mutation prediction strategies are combined to further obtain CCT mutants with significantly improved performance, so as to provide a core catalytic element for efficient biosynthesis of cytidine diphosphate choline.

This invention provides a highly toxic Cryotoxin protein obtained through AI-driven rational design and its applications. By leveraging artificial intelligence for precise prediction and optimized design of domain functions, the Cryotoxin mutant with domain replacement can more efficiently recognize intestinal receptors of target pests, significantly enhancing its toxic activity against the lepidopteran cotton bollworm. The domain reconstruction and optimization alters the binding mode between the original toxin and the pest receptor, reducing the possibility of pests evading the toxic effect through a single resistance mechanism and extending the field lifespan of the mutant. The Cryotoxin mutant provided by this invention, as a highly efficient and safe biocontrol material, can replace some chemical pesticides, reducing the impact of pesticide residues on agricultural product quality and lowering pollution to soil, water, and other ecological environments; it also improves crop yield and quality, promoting green and sustainable agricultural development, and possessing significant ecological and economic benefits.

The application relates to the technical field of material science, solves the technical problem that traditional methods cannot efficiently and accurately predict the mechanical properties of high-entropy alloys, and cannot reveal the performance influence mechanism and provide clear guidance for actual process optimization, and particularly relates to a high-entropy alloy mechanical property prediction and optimization method based on an artificial neural network, a mechanical property data set of a target high-entropy alloy under multiple working condition parameters is constructed; an artificial neural network model for mapping the nonlinear relationship between the working condition parameters and the mechanical properties is constructed and trained and optimized and verified, the influence degree of each working condition parameter on the mechanical property prediction is determined based on feature importance analysis, and reverse optimization of the working condition parameters is carried out to obtain optimal working condition parameters meeting target mechanical properties. The application realizes efficient and accurate prediction of the mechanical properties of high-entropy alloys and optimization of process parameters, the prediction efficiency is improved by 4-5 orders of magnitude compared with molecular dynamics simulation, and technical support is provided for rational design of high-entropy alloys.

De novo protein design, the rational design of new proteins from scratch with specific functions and properties, is a grand challenge in molecular biology. Recently, deep generative models have emerge