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A method for producing heat-stable and pure trypsin

ABSTRACT: In this study we evaluated several different strategies for reductive methylation and purification of porcine trypsin in order to write a protocol that can be followed by proteomics laboratories.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Equus Caballus (horse) Homo Sapiens (human) Bos Taurus (bovine)

SUBMITTER: Søren Heissel

LAB HEAD: Peter Højrup

PROVIDER: PXD008193 | Pride | 2019-06-06

REPOSITORIES: Pride

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Publications

Enhanced trypsin on a budget: Stabilization, purification and high-temperature application of inexpensive commercial trypsin for proteomics applications.

Heissel Søren S Frederiksen Sigurd J SJ Bunkenborg Jakob J Højrup Peter P

PloS one 20190627 6

Trypsin is by far the most commonly used protease in proteomics. Even though the amount of protease used in each experiment is very small, digestion of large amounts of protein prior to enrichment can be rather costly. The price of commercial trypsin is highly dependent on the quality of the enzyme, which is determined by its purity, activity, and chemical modifications. In this study we evaluated several strategies for improving the quality of crude trypsin by reductive methylation and affinity ...[more]

PMID: 31246970

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Influence of Asp Isomerization on Trypsin and Trypsin-like Proteolysis.

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