Lysozyme, hydrolysis

The initial step in the alternative hydrolysis mechanism is protonation of the ring 0i by Glu 35 (Scheme I). Cleavage of the endocyclic C1-O5 bond forms the acyclic oxocarbonium ion intermediate, which is stabilized by Asp 52. Attack by water, cleavage of the C1-O4 bond, and ring closure then lead to the observed products. Existing experimental data on lysozyme hydrolysis are consistent with Scheme I (see references in Post and Karplus ( )). Moreover, distortion of the ring in site D is not required and the antiperiplanar orientation of an exocyclic O4 lone pair orbital relative to the cleaved C1-O5 bond found in the simulation (see section on "Enhancement of a Substrate Conformation Optimum for Catalysis") is in accord with stereoelectronic requirements (1 ). ... 

Additional studies of the enzyme-substrate complementarity in other complexes along the reaction path are under way. Since the initial report of an alternative pathway for lysozyme hydrolysis (8,9.28) work on the solution hydrolysis of glucosides has demonstrated the existence of a ring opening mechanism (29,30). ... 

Lysozyme Hydrolysis of cell-wall polysaccharides of various bacterial species leading to the breakdown of the cell wall most often in Gram-positive bacteria. 

Post, C. B., Dobson, C. M. and Karplus, M. (1990). Lysozyme hydrolysis of P-glycosides a consensus between binding interactions and mechanisms. ACS Symposium Series, chapter 23. 

Aiba, S. 1992. Studies on chitosan 4. Lysozymic hydrolysis of partially A-acetylated chitosans. International Journal of Biological Macromolecules 14 225-228. 

S. Aiba, Studies on chitosan Lysozymic hydrolysis of partially N-acetylated chitosans, Int J. Biol Macromol, 14 (4), 225-228,1992. 

Hydrolysis Rate Constants for Model Oligosaccharides with Lysozyme... 

The mechanism of the lysozyme reaction is shown in Figures 16.36 and 16.37. Studies using O-enriched water showed that the Ci—O bond is cleaved on the substrate between the D and E sites. Hydrolysis under these conditions incorporates into the Ci position of the sugar at the D site, not into the oxygen at C4 at the E site (Figure 16.36). Model building studies place the cleaved bond approximately between protein residues Glu and Asp. Glu is in a nonpolar or hydrophobic region of the protein, whereas Asp is located in a much more polar environment. Glu is protonated, but Asp is ionized... 

Hen egg-white lysozyme catalyzes the hydrolysis of various oligosaccharides, especially those of bacterial cell walls. The elucidation of the X-ray structure of this enzyme by David Phillips and co-workers (Ref. 1) provided the first glimpse of the structure of an enzyme-active site. The determination of the structure of this enzyme with trisaccharide competitive inhibitors and biochemical studies led to a detailed model for lysozyme and its hexa N-acetyl glucoseamine (hexa-NAG) substrate (Fig. 6.1). These studies identified the C-O bond between the D and E residues of the substrate as the bond which is being specifically cleaved by the enzyme and located the residues Glu 37 and Asp 52 as the major catalytic residues. The initial structural studies led to various proposals of how catalysis might take place. Here we consider these proposals and show how to examine their validity by computer modeling approaches. 

LD model, see Langevin dipoles model (LD) Linear free-energy relationships, see Free energy relationships, linear Linear response approximation, 92,215 London, see Heitler-London model Lysine, structure of, 110 Lysozyme, (hen egg white), 153-169,154. See also Oligosaccharide hydrolysis active site of, 157-159, 167-169, 181 calibration of EVB surfaces, 162,162-166, 166... 

Because 0-acyl chitins appear to be scarcely susceptible to lysozyme, the susceptibility of DBG to Upases has been studied to obtain insight into its biodegradability in vivo. The changes in infrared and X-ray diffraction spectra of the fibers support the slow degradation of DBG by Upases [125,126]. The chemical hydrolysis of DBG to chitin is the most recent way to produce regenerated chitin. 

In most cases the microspheres were insoluble. The polysaccharides might be partially cross-linked via amido groups formed by the carboxyl groups of the polyanion and the restored free amino group of chitosan. The susceptibility to enzymatic hydrolysis by lysozyme was poor, mainly because lysozyme, a strongly cationic protein, can be inactivated by anionic polysaccharides [207]. 

Procarboxypeptidase A is activated by the removal of a peptide of some 64 residues from the N-terminus by trypsin.153 This zymogen has significant catalytic activity. As well as catalyzing the hydrolysis of small esters and peptides, procarboxypeptidase removes the C-terminal leucine from lysozyme only seven times more slowly than does carboxypeptidase. Also, the zymogen hydrolyzes Bz-Gly-L-Phe with kcsA = 3 s-1 and KM = 2.7 mM, compared with values of 120 s 1 and 1.9 mM for the reaction of the enzyme.154 In contrast to the situation in chymotrypsinogen, the binding site clearly pre-exists in procarboxypeptidase, and the catalytic apparatus must be nearly complete. 

Highly-efficient general acid catalysis of acetal hydrolysis is involved in the reactions of glycosidase enzymes such as lysozyme (Dunn and Bruice, 1973)... 

Tests allowing precise determination of the conditions to protect the soluble protein, and of the temperature at which the reaction was slow enough for X-ray data collection, were sought. To ascertain the best conditions for the determination of the structure of a productive lysozyme-substrate complex, the hydrolysis of bacterial cell walls and oligosaccharides was investigated both in high-salt solutions and in mixed solvents. 

---