Biosynthesis of epsilon-poly-L-lysine in a cell-free system of Streptomyces albulus.
epsilon-Poly-L-lysine (epsilon-PL) is a homo-poly-amino acid characterized by a peptide bond between carboxyl and epsilon-amino groups of L-lysine. Here we report the cell-free synthesis of epsilon-PL by a sensitive radioisotopic epsilon-PL assay system. In vitro epsilon-PL synthesis depended on ATP and was not affected by ribonuclease, kanamycin, or chloramphenicol. epsilon-PL synthesizing activity was detected in the membrane fraction. The reaction product, epsilon-PL, from L-lysine was identified by MALDI-TOF MS and the number of lysine residues of the epsilon-PL products was apparently 11-34. These results suggest that the biosynthesis of epsilon-PL is nonribosomal peptide synthesis and is catalyzed by membrane bound enzyme(s). The enzyme preparation showing the epsilon-PL synthesizing activity also catalyzed lysine-dependent AMP production and an ATP-PPi exchange reaction, suggesting that L-lysine is adenylated in the first step of epsilon-PL biosynthesis.
Kawai
Takahiro
T
Department of Biotechnology, Faculty of Engineering, Tottori University, Koyama-minami, Tottori 680-8552, Japan.
Kubota
Takaaki
T
Hiraki
Jun
J
Izumi
Yoshikazu
Y
D000249:Adenosine Monophosphate / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002474:Cell-Free System; D002701:Chloramphenicol / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D007612:Kanamycin / Q000494:pharmacology; D008239:Lysine / Q000737:chemistry; D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry; D010710:Phosphates / Q000737:chemistry; D011107:Polylysine / Q000096:biosynthesis*; D012260:Ribonucleases / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013302:Streptomyces / Q000378:metabolism*; D013997:Time Factors
D010455:Peptides; D010710:Phosphates; D011107:Polylysine; D000249:Adenosine Monophosphate; D007612:Kanamycin; D002701:Chloramphenicol; D000255:Adenosine Triphosphate; D012260:Ribonucleases; D008239:Lysine