| Résumé | Liquid chro ma tog ra phy–mass spec trom e try was applied to deter mine the action pattern of dif fer ent chon droi tin lyases. Two com mer cial enzymes, chon dro itin ase ABC (Pro teus vul ga ris) and chon dro itinase ACII (Arth ro bac ter au res cens), hav ing action pat terns pre vi ously deter mined by vis cos i me try and gel elec tro pho re sis were first exam ined. Next, the action pat terns of recombinant lyases, chon dro itin ase ABC from Bac te roi des theta i ota omi cron (expressed in Esch e richia coli) and chon dro itin ase AC from Fla vo bac terium hep ar i num (expressed in its original host), were exam ined. Chon droi tin sul fate A (CS-A, also known as chon droi tin-4-sul fate) was used as the sub strate for these four lyases. Ali quots taken at var i ous time points were ana lyzed. The prod ucts of chon dro itin ase ABC (P. vul ga ris) and chon dro itin ase AC (F. hepar i num) con tained unsat u rated oli go sac cha rides of sizes rang ing from disac cha ride to deca sac cha ride, dem on strat ing that both are end o lyt ic enzymes. The prod ucts affor ded by chon dro itin ase ABC (B. thetai ota omi cron) and chon dro itin ase ACII (A. au res cens) con tained pri mar ily unsat u rated disac cha ride. These two ex o lyt ic enzymes showed dif fer ent minor prod ucts, sug gest ing some sub tle spec i fic ity dif fer ences between the actions of these two ex o lyt ic lyases on chon droi tin sul fate A. |
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