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Literature summary for 3.2.1.17 extracted from

  • Cancado, F.C.; Barbosa, J.A.; Marana, S.R.
    Role of the triad N46, S106 and T107 and the surface charges in the determination of the acidic pH optimum of digestive lysozymes from Musca domestica (2010), Comp. Biochem. Physiol. B, 155, 387-395.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of wild-type and mutant MdL1 and MdL2 in Pichia pastoris Musca domestica

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant MdL2, sitting drop vapour diffusion method, 18°C, precipitant solution containing 28% isopropanol, 21% PEG 4000, and 0.115 M sodium citrate pH 4.2, addition of 0.2 M ammonium acetate, 30% 2-methyl-2,2-pentanediol, 0.1 M sodium citrate, pH 5.6, X-ray diffraction structure determination and analysis at 1.9 A resolution, modeling Musca domestica

Protein Variants

Protein Variants Comment Organism
additional information introduction of six basic residues onMdL1 surface increases by 1 unit the pH optimum for the activity upon bacterial walls Musca domestica
N46D/S106V/T107A site-directed mutagenesis of isozyme MdL2, the mutant pKas are increased compared to the wild-type pKas Musca domestica

Organism

Organism UniProt Comment Textmining
Musca domestica
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digestive lysozymes 1 and 2, i.e. MdL1 and MdL2
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant MdL2s from Pichia pastoris by ammonium sulfate fractionation, and cation exchange, for the sextuple mutant, or anion exchange, for the other mutants, chromatography Musca domestica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-methylumbelliferyl-beta-D-N,N',N''-triacetylchitotrioside + H2O
-
Musca domestica ?
-
?

Subunits

Subunits Comment Organism
More structure analysis of MdL1 and MdL2, overview. Residues S106-T107 delimit a polar pocket around E32, as catalytic acid/base, and N46 contributes to the positioning of D50, as catalytic nucleophile Musca domestica

Synonyms

Synonyms Comment Organism
Mdl1
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Musca domestica
MdL2
-
Musca domestica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
additional information
-
the acidic pH optimum for MdL2 and MdL1 activities upon methylumbelliferylchitotrioside is determined by the presence of N46, S106 and T107 in the environment of their catalytic residues, which favors pKas reduction. The acidic pH optimum upon bacterial walls is determined by a low concentration of positive charges on the MdL2 and MdL1 surfaces Musca domestica

General Information

General Information Comment Organism
additional information comparison of the structure, surface charge, dissociation constants, and pH optimum of the fly enzyme with the enzyme from egg white, overview Musca domestica