Recombinant Engineered TEV Protease-AI is a genetically optimizedform of Tobacco Etch Virus (TEV) nuclear-inclusion-a endopeptidasedesigned for enhanced cleavage efficiency at the canonical seven–amino acid TEV recognition sequence: Glu-Asn-Leu-Tyr-Phe-Gln↓(Gly/Ser).The protease exhibits high site specificity and catalytic efficiency,making it suitable for the removal of affinity tags from purified recombinant proteins. AiTEV Protease demonstrates efficient cleavageat room temperature (25–35°C) across a broad range of buffercompositions (pH 4.0–9.0), with maximal activity observed at pH 8.0and 30°C. Compared to the native TEV protease, AiTEV Protease exhibits improved stability, higher activity, and greater tolerance to buffer and temperature variations.