Protease Protein

Protease K is derived from Tritirachium album Limber. It non-specifically cuts the carboxy-terminal peptide bonds of aliphatic amino acids and aromatic amino acids, and has a strong ability to degrade natural proteins, and has a high activity in the buffer solution for extracting DNA and RNA. It can be used for the separation of plasmid or genomic DNA and RNA, and is a key reagent for DNA extraction.

SUMO protease, also known as Ulp, is a highly active cysteine protease that recognizes the tertiary structure of SUMO protein, rather than the amino acid sequence, and thus can efficiently and specifically cut SUMO protein from the recombinant fusion protein. Compared to the short recognition sites of EK and TEV, SUMO protease can recognize the complete SUMO structure, so SUMO protease does not miscut fused proteins. The effectiveness of SUMO protease can be maintained in a wide range of reaction environments, such as temperature (4-30℃), pH (5.5-9.5), etc. SUMO proteases also have a poly His label, which facilitates affinity chromatography purification of the fusion protein after cleavage.

Recombinant TEV protease was expressed and purified by genetic engineering. TEV protease specifically recognizes the seven amino acid sequences composed of Glu-Asn-Leu-Tyr-Phe-Gly/Ser and cuts between Gln and Gly/Ser. Compared with EK, SUMO and other proteases, TEV protease has high activity and high specificity. Tev protease was purified by 6×His label (including histamine acid label), and its purity was up to 99%. After the enzyme shear reaction is complete, it can be removed by His label purification Resin Ni-NTA resin, so as to achieve the purpose of purification of the target protein. It is active at 4-30℃ and pH 6.0-8.5, and the reaction conditions are relatively loose. It is mainly used for fusion protein label removal. This protein is derived from recombinant expression of Escherichia coli.

HRV 3C protease is a common tool enzyme used to remove affinity labels from fusion proteins. Cysteine protease that recognizes the cleavage site of Leu-Glu-Val-Leu-Phe-Gln↓-Gly-Pro has histidine label and is easily removed by Ni-NTA resin.

EK Protease is a common tool enzyme used to remove the affinity label on the fusion protein. It can identify the protease with five amino acid sequence DDDDK↓, and no excess amino acid remains after enzyme digestion.

Caspase-2 is a specific cutting enzyme involved in the activation cascade of Caspase, which is responsible for the execution of apoptosis. May work by activating proteins needed for cell death or deactivating proteins needed for cell survival. The cleavage site is the N-terminal residue of target protein (POI). It can complete 85-90% cutting in 15 minutes.