| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | The putative chondroitinase ACII from Arthrobacter sp. 161MFSha2.1, hereafter referred to as tA16ACII, was synthesized without its N-terminal signal peptide and cloned into the NdeI and XhoI sites of pET28a(+) (GenScript) in frame with the N-terminal 6x-His tag for purification (the strains and plasmids used in this study are provided in Table S1). The nucleotide sequence, excluding the 33 N-terminal residues forming the predicted signal peptide, was synthesized (express cloning option) into NdeI and XhoI restriction sites of pET-28a(+) by GenScript (Piscataway, NJ), creating an N-terminal 6xHis-tag fusion for expression in E. coli as shown in Figure S7. | Get A Quote |
Chondroitin sulfates are the glycosaminoglycan chains of proteoglycans critical in the normal development and pathophysiology of all animals. Chondroitinase ACII, a polysaccharide lyase originally isolated from Arthrobacter aurescens IAM 110 65, which is widely used in the analysis and study of chondroitin structure, is no longer commercially available. The aim of the current study was to prepare recombinant versions of this critical enzyme for the glycobiology research community. Two versions of recombinant chondroitinase ACII were prepared in Escherichia coli, and their activity, stability, specificity and action pattern were examined, along with a non-recombinant version secreted by an Arthrobacter... More
