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Published on: April 28, 2022
Streptomyces erythraeus trypsin for proteomics applications.
Jianying Z Kiser1, Marc Post, Benlian Wang
1Case Center for Proteomics and Bioinformatics, Department of Pharmacology, Department of Ophthalmology and Visual Sciences, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, Ohio, USA.
Journal of Proteome Research
|February 24, 2009
Summary
Streptomyces erythraeus trypsin (SET) demonstrates superior amidase activity and resistance to degradation compared to mammalian trypsin. Recombinant SET (rSET) shows promise for proteomics applications, including protein digestion and peptide labeling.
Area of Science:
- Biochemistry
- Proteomics
- Enzymology
Background:
- Mammalian trypsins (bovine, porcine) are standard for proteomics.
- Alternative trypsin sources offer enhanced catalytic activity and stability.
- Streptomyces erythraeus trypsin (SET) exhibits significantly higher amidase activity and autolytic resistance.
Purpose of the Study:
- To evaluate the potential of recombinant Streptomyces erythraeus trypsin (rSET) for proteomics.
- To characterize the enzymatic properties of rSET.
- To assess rSET performance in standard proteomics workflows.
Main Methods:
- Expression of SET in E. coli and purification to homogeneity.
- Enzymatic assays measuring amide bond hydrolysis activity.
- Application of rSET for in-solution and in-gel digestion of bovine serum albumin.
- Evaluation of rSET for 18O peptide labeling.
Main Results:
- rSET displayed over an order of magnitude higher amide bond hydrolysis activity than modified porcine trypsin.
- rSET showed no autolytic degradation after 96 hours of incubation at 37°C.
- rSET effectively performed protein digestion and 18O labeling, confirming its utility.
Conclusions:
- Recombinant Streptomyces erythraeus trypsin (rSET) is a highly active and stable protease.
- rSET demonstrates significant potential as a valuable tool in proteomics research.
- Further characterization of rSET properties supports its application in diverse proteomic experiments.
