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Peptide Synthesis of Aspartame Precursor Using Organic‐Solvent‐Stable PST‐01 Protease in Monophasic Aqueous‐Organic Solvent Systems
Author(s) -
Tsuchiyama Shotaro,
Doukyu Noriyuki,
Yasuda Masahiro,
Ishimi Kosaku,
Ogino Hiroyasu
Publication year - 2007
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp060382y
Subject(s) - aspartame , chemistry , aqueous solution , ethylene glycol , yield (engineering) , phenylalanine , solvent , organic chemistry , protease , medicinal chemistry , methanol , nuclear chemistry , enzyme , amino acid , biochemistry , materials science , metallurgy
Abstract The PST‐01 protease is a metalloprotease that has zinc ion at the active center and is very stable in the presence of water‐soluble organic solvents. The reaction rates and the equilibrium yields of the aspartame precursor N ‐carbobenzoxy‐ l ‐aspartyl‐ l ‐phenylalanine methyl ester (Cbz‐Asp‐Phe‐OMe) synthesis from N ‐carbobenzoxy‐ l ‐aspartic acid (Cbz‐Asp) and l ‐phenylalanine methyl ester (Phe‐OMe) in the presence of water‐soluble organic solvents were investigated under various conditions. Higher reaction rate and yield of Cbz‐Asp‐Phe‐OMe were attained by the PST‐01 protease when 30 mM Cbz‐Asp and 500 mM Phe‐OMe were used. The maximum reaction rate was obtained pH 8.0 and 37 °C. In the presence of dimethylsulfoxide (DMSO), glycerol, methanol, and ethylene glycol, higher reaction rates were obtained. The equilibrium yield was the highest in the presence of DMSO. The equilibrium yield of Cbz‐Asp‐Phe‐OMe using the PST‐01 protease attained 83% in the presence of 50% (v/v) DMSO.