Glycosylation kinetics

A quantitative interpretation of aldonolactone inhibition in terms of an adaptation of the active site to a transition state approaching a planar, glycosyl oxocarbonium ion is made difficult for several reasons. Due to the interconversion between the 1,4- and 1,5-lactones, and their hydrolysis to the aldonic acids, their use is limited to kinetic studies with incubation times of 10 min or less. This was not realized by most investigators prior to 1970. In many cases, only the 1,4-lactone can be isolated its (partial) conversion into... 

Hehre and coworkers showed that beta amylase from sweet potatoes, an inverting, a-specific exo-(l 4)-glucanase, catalyzes the hydrolysis of jS-maltosyl fluoride with complex kinetics which indicated the participation of two substrate molecules in the release of fluoride ion. Furthermore, the reaction was strongly accelerated by the addition of methyl ) -maltoside. Hydrolysis of a-maltosyl fluoride, on the other hand, obeyed Michaelis-Menten kinetics. The main product with both a- and yj-maltosyl fluoride was )S-maltose. The results with )3-maltosyl fluoride were interpreted by the assumption of a glycosylation reaction preceding hydrolysis by which a malto-tetraoside is formed by the replacement of fluoride ion by a second substrate molecule or added methyl -maltoside (see Scheme 5). 

T. A. Gerken, Kinetic modeling confirms the biosynthesis of mucin core 1 (/i-Gal (1 -3)x-Gal YAc-O-Ser/Thr) O-glycan structures are modulated by neighboring glycosylation effects, Biochemistry, 43 (2004) 4137—4142. 

Conventionally, the use of NaH or Cs2C03 as a base for the reaction of glycosyl hemiacetals with C13CCN often yields the thermodynamically favored a-glycosyl trichloroacetimidates, whereas the use of K2C03 often yields kinetically controlled P-glycosyl trichloroacetimidates (Scheme 3.26). The use of l,8-diazabicyclo[5.4.0] undec-7-ene (DBU) often provides a/p mixtures, mostly favoring a-products. 

Other anions such as nitronate anion,96 or the conjugate base of (3-ketoesters,97 give similar results. Usually, the 1,2-cw-C-glycosyl compound is formed under kinetic conditions. Thus, in the case of the reaction of the GlcNAc derivative 75 with the Wittig reagent Ph3P=CHCOOEt, the a-C-pyranosyl compound 77a is the major product under conditions of kinetic control. Under basic conditions, 77a is slowly epimerized into the more stable (3-C-pyranosyl derivative 77p (Scheme 25). 

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