Description:
<jats:p>The substrate kinetics of chitin synthase (CS) were non‐Michaelian, irrespective of the type of enzyme preparation studied (105‐S chitosomal CS, and 16‐S CS <jats:italic>ex</jats:italic> walls), even in the presence of saturating GlcNAc. An unexplained idiosyncrasy of this enzyme, which is likely to be responsible for this phenomenon, is evident from the striking non‐linearity of product deposition with time at low substrate or low enzyme concentrations, particularly in the absence of GlcNAc. The possibility can be excluded that this non‐linearity is due to the formation of soluble by‐products or intermediates in the form of chito‐oligomers, as shown by HPLC/pulsed amperometric detection analysis. Additional evidence was sought for the tenet that CS is homotropically‐heterotropically regulated, at least under steady‐state reaction conditions. Substrate kinetic curves established from rate data for the linear reaction phase only were used for modelling. These could be reasonably well parameterised on the basis of the Monod mathematical model for co‐operative ligand binding. Within a series of test compounds used to assess the stereochemical conditions of the allosteric site of CS for effector binding, <jats:italic>N</jats:italic>‐acetylglucosaminono‐1,5‐lactone oxime excelled. Requirements for effector binding are as follows: (a) an aminoglucopyranose skeleton with the amino function acetylated, and (b) a single‐bonded oxo‐function present at C(1), which is preferentially a hydrogen bond donor, that may be equatorially spaced off, but must not be α‐anomeric. The implications of these findings for chitin synthesis <jats:italic>in vivo</jats:italic> are discussed in terms of a mechanistically based fitness of CS to operate efficiently under vastly different combinations of substrate and effector concentrations as well as with respect to the conditions under which chitin synthase could be coordinately linked to the catabolism of chitin.</jats:p>