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Cloning and Expression of Recombinant Chondroitinase ACII and Its Comparison to the Arthrobacter aurescens Enzyme
Author(s) -
Williams Asher,
He Wenqin,
Cress Brady F.,
Liu Xinyue,
Alexandria Jordanne,
Yoshizawa Hiroki,
Nishimura Kazuhiro,
Toida Toshihiko,
Koffas Mattheos,
Linhardt Robert J.
Publication year - 2017
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201700239
Subject(s) - recombinant dna , arthrobacter , biochemistry , chondroitin sulfate , enzyme , chondroitin , escherichia coli , chemistry , glycosaminoglycan , biology , gene
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 is to prepare recombinant versions of this critical enzyme for the glycobiology research community. Two versions of recombinant chondroitinase ACII are prepared in Escherichia coli , and their activity, stability, specificity, and action pattern are examined, along with a non‐recombinant version secreted by an Arthrobacter strain. The recombinant enzymes are similar to the enzyme obtained from Arthrobacter for all examined properties, except for some subtle specificity differences toward uncommon chondroitin sulfate substrates. These differences are believed to be due to either post‐translational modification of the Arthrobacter ‐secreted enzyme or other subtle structural differences between the recombinant and natural enzymes. The secreted chondroitinase can serve as a suitable replacement for the original enzyme that is currently unavailable, while the recombinant ones can be applied generally in the structural determination of most standard chondroitin sulfates.