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Mutagenesis Services> | … Materials and methods Protein expression and purification The gene encoding Sm-SrtE2 without N-terminal anchor was codon-optimized by GenScript (New Jersey, USA) and inserted into pET28a vector. Mutagenesis for exchange of glutamines for … | Get A Quote |
Distinct streptomycetes such as Streptomyces mobaraensis produce the protein cross-linking enzyme transglutaminase. Bioinformatic analysis predicted the occurrence of seven sortases exerting transpeptidation reactions similarly to transglutaminase. Here, we report the production and characterization of sortase E2 (Sm-SrtE2) solubilized by removal of its membrane anchor domain. Sm-SrtE2 activity was measured using pentapeptides predicted to be cell wall sorting signals of putative sortase substrate proteins. Preferred linkage to Gly by Sm-SrtE2 was in the order LAETG>LAHTG>LAQTG~LANTG>LARTG. Chaplin 1 from S. mobaraensis was further demonstrated to be an excellent substrate of both the intrinsic Sm-SrtE2 and... More