Enzymatic synthesis of β-N-(γ-L(+)-glutamyl)phenylhydrazine with Escherichia coli γ-glutamyltranspeptidase.
A new method for the synthesis of β-N-(γ-L(+)-glutamyl)phenylhydrazine is presented. This compound was prepared from L-glutamine and phenylhydrazine through a transpeptidation reaction of Escherichia coli γ-glutamyltranspeptidase although phenylhydrazine has been reported to be an inhibitor of the enzyme. The optimum reaction conditions were 60 mM L-glutamine, 300 mM phenylhydrazine, 40 U γ-glutamyltranspeptidase/ml, and pH 9 in approx. 800 ml. After 6 h at 37 °C, the product was obtained with a conversion rate of 93 % (mol/mol). γ-Glutamyltranspeptidase was reversibly inhibited only when phenylhydrazine was above 300 mM.
State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing, 210093 Jiangsu, China.
Hongjuan
H
Zhang
Yueping
Y
Zhan
Junjun
J
Chang
Junzhong
J
Liu
Lisheng
L
Xu
Zhiyuan
Z
Wang
Qian
Q
Liu
Qingcai
Q
Jiao
D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005971:Glutamates / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D010659:Phenylhydrazines / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature; D005723:gamma-Glutamyltransferase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D029968:Escherichia coli Proteins; D005971:Glutamates; D010659:Phenylhydrazines; D011994:Recombinant Proteins; D005723:gamma-Glutamyltransferase