Tianjin University

Tianjin University of Science and Technology

Shandong Lonct Enzymes Co. Ltd

Fufeng Liu

Yan Sun

Yu Li

Fuping Lu

Wenlong Liu

Qinghong Shi

Xingji Wang

Kefen Wang

Weiwei Zhao

Yihan Liu

Huitu Zhang

An alkaline protease mutant, and a gene, engineered strain, a preparation method and application thereof are provided. The method comprises the following steps of extracting genome DNA of, performing PCR amplification to obtain a wild-type alkaline protease gene sequence, mutating the wild-type alkaline protease gene obtained by the amplification through an error-prone PCR, performing high-throughput screening to obtain a plurality of highly active alkaline protease genes, performing DNA shuffling on the highly active alkaline protease genes, and performing screening to obtain eight alkaline protease mutant genes with higher activity.

Alkaline protease mutant, and gene, engineered strain, preparation method and application thereof

An affinity ligand peptide library of IgG constructed on the basis of Protein A affinity model and the application of a design method thereof. According to the Molecular Mechanics&#x2014;Poisson-Boltzmann surface area (MM/PBSA) method and on the basis of the known human IgG-Protein A complex structure, the hot spots of Protein A that have high affinity for human IgG are obtained analytically, and a Protein A simplified affinity model is built thereof. An affinity peptide library of IgG is constructed including heptapeptide and octapeptide structural modes. On the basis of the peptide structural modes, the types of inserted amino acids that 'X' residues represent are further identified using amino acid location method. Then, molecular docking and molecular dynamics simulation methods are used to screen the candidate peptides successively. Finally, the affinity peptide ligands that can effectively purify IgG are identified using affinity chromatography.

Method for constructing a novel affinity peptide library for binding immunoglobulin G based on a protein affinity model of protein A

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Tianjin, China

Fufeng Liu