Cloning and expression of recombinant chondroitinase AC II and its comparison to the Arthrobacter aurescens enzyme
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Authors
Williams, A.
He, W.
Cress, B.F.
Liu, X.
Alexandria, J.
Yoshizawa, H.
Nishimura, Kazuhiro
Toida, T.
Koffas, M.
Linhardt, Robert J.
Issue Date
2017
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Alternative Title
Abstract
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.
Description
Biotechnology Journal, 12, 1700239
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Full Citation
Cloning and expression of recombinant chondroitinase AC II and its comparison to the Arthrobacter aurescens enzyme, A. Williams, W. He, B. F. Cress, X. Liu, J. Alexandria, H. Yoshizawa, K. Nishimura, T. Toida, M. Koffas, R. J. Linhardt, Biotechnology Journal, 12, 1700239, 2017.
Publisher
Wiley