Expression and functional analysis of high-stability hybrid human trypsin
Xiaodong Cui1, Xiaohong Tian1, Chenyan Li1
1Institute of Biotechnology, Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan 030006, China; Biomedical and Health Laboratory in Shanxi Province, Taiyuan 030006, China.
Enzyme and Microbial Technology
|May 7, 2026
Summary
Researchers engineered a novel recombinant human hybrid trypsin (rhhT) by combining properties of natural trypsins. This rhhT enzyme shows enhanced stability and maintained catalytic activity for industrial applications.
Area of Science:
- Biochemistry
- Enzymology
- Protein Engineering
Background:
- Trypsin is vital for industrial processes, but natural forms have limitations.
- Enhancing trypsin's stability and activity is crucial for optimizing production efficiency.
Purpose of the Study:
- To engineer a recombinant human hybrid trypsin (rhhT) with improved stability and retained catalytic function.
- To evaluate the performance of rhhT for potential industrial applications.
Main Methods:
- Constructed rhhT by integrating features of human anionic trypsin (hT2) and cationic trypsin (hT1).
- Expressed and purified rhhT using inclusion body formation in E. coli, ion exchange chromatography, and denaturation-refolding.
- Characterized rhhT's enzymatic properties, including optimal conditions, kinetics, and stability under various stresses.
Main Results:
- Purified rhhT demonstrated significantly enhanced stability compared to natural trypsin.
- rhhT maintained catalytic activity comparable to rhT2, with no significant differences in Km or Kcat.
- rhhT showed superior resistance to temperature, pH variations, ionic stress, degradation, and improved storage stability, with distinct calcium ion binding.
Conclusions:
- Successfully designed a "highly activated and stable" human hybrid trypsin (rhhT).
- rhhT offers a robust foundation for industrial applications in medical and food processing.
- The engineered trypsin overcomes limitations of natural variants, paving the way for improved bioprocessing.
