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Literature summary for 3.4.24.27 extracted from

  • Birrane, G.; Bhyravbhatla, B.; Navia, M.
    Synthesis of aspartame by thermolysin: an x-ray structural study (2014), ACS Med. Chem. Lett., 5, 706-710.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.24.27

Application

Application Comment Organism
synthesis using the enzyme to catalyze the condensation of the chiral aspartame-precursor, carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester, from the protected amino acid substrates carbobenzoxy-L-aspartic acid and L-phenylalanine methyl ester in large scale production. Analysis of the protease mediated peptide synthesisof a precursor of the artificial sweetener aspartame, a multiton peptide synthesis catalyzed by the enzyme thermolysin. X-ray structures of thermolysin in complex with aspartame substrates separately, and after protease mediated peptide synthesis in a crystal, rationalize the reaction's substrate preferences and reveal an unexpected form of substrate inhibition that explains its sluggishness. Structure guided optimization of this and other PMPS reactions could expand the economic viability of commercial peptides beyond current high-potency, low-volume therapeutics, with substantial green chemistry advantages Bacillus thermoproteolyticus

Crystallization (Commentary)

Crystallization (Comment) Organism
analysis of the crystal structure of enzyme complexed with substrates L-phenylalanine methyl ester and carbobenzoxy-L-aspartic, or with inhibitor carbobenzoxy-L-phenylalanine-phosphonamidate-L-leucyl-L-alanine, X-ray diffraction structure determination and analysis, modeling Bacillus thermoproteolyticus

Inhibitors

Inhibitors Comment Organism Structure
carbobenzoxy-D-aspartic acid
-
 Bacillus thermoproteolyticus
carbobenzoxy-L-aspartic acid substrate inhibition Bacillus thermoproteolyticus
carbobenzoxy-L-phenylalanine-phosphonamidate-L-leucyl-L-alanine a potent phosphonamidate transition state analogue inhibitor Bacillus thermoproteolyticus
additional information binding of inhibitors to the active site of thermolysin, structure overview Bacillus thermoproteolyticus
N-(1-carboxy-3-phenyl-propyl)-L-leucyl-L-tryptophan
-
 Bacillus thermoproteolyticus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information steady-state kinetics, overview Bacillus thermoproteolyticus

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
 Bacillus thermoproteolyticus
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ a Zn2+ metalloprotease Bacillus thermoproteolyticus

Organism

Organism UniProt Comment Textmining
Bacillus thermoproteolyticus P00800
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
carbobenzoxy-L-aspartic acid + L-phenylalanine methyl ester condensation, the enzyme is enantioselective for the desired L-phenylalanine methyl ester substrate from a racemic mixture of DL-phenylalanine methyl ester. In contrast, although both enantiomers of carbobenzoxy-L-aspartic acid can bind to the enzyme, only carbobenzoxy-L-aspartic acid is used since carbobenzoxy-D-aspartic acid inhibits the enzyme, substrate carbobenzoxy-L-aspartic acid binding structures, detailed overview Bacillus thermoproteolyticus carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester precipitation as the water-insoluble Phe-OMe salt drives the overall reaction in the direction of peptide synthesis ?
additional information binding of substrates to the active site of thermolysin, overview Bacillus thermoproteolyticus ?
-
 ?

Synonyms

Synonyms Comment Organism
TLN
-
 Bacillus thermoproteolyticus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000000068
-
 carbobenzoxy-L-phenylalanine-phosphonamidate-L-leucyl-L-alanine pH and temperature not specified in the publication Bacillus thermoproteolyticus
0.00005
-
 N-(1-carboxy-3-phenyl-propyl)-L-leucyl-L-tryptophan pH and temperature not specified in the publication Bacillus thermoproteolyticus

General Information

General Information Comment Organism
additional information proposed mechanism of action for the thermolysin-catalyzed synthesis of aspartame precursor carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester, overview. Residues E143, Y157, and H231 represent the catalytic triad, enzyme active site structure analysis Bacillus thermoproteolyticus