Transfer of D-Phenylalanine from Gramicidin S Synthetase 1 to Gramicidin S Synthetase 2 in Gramicidin S Synthesis

• Published in: Journal of biochemistry (Tokyo) 1983-01, Vol.93 (1), p.177-188
• Main Authors: HORI, Kazuko, KANDA, Masayuki, MIURA, Setsuko, YAMADA, Yoko, SAITO, Yoshitaka
• Format: Article
• Language: English
• Subjects: Amino Acid Isomerases - analysis; Bacillus - enzymology; Bacillus - genetics; Bacillus brevis; Bacteriology; Binding Sites; Biological and medical sciences; Chromatography, Thin Layer; Fundamental and applied biological sciences. Psychology; Gramicidin - biosynthesis; Metabolism. Enzymes; Microbiology; Multienzyme Complexes - analysis; Mutation; Peptide Synthases - analysis; Phenylalanine - metabolism; Structure-Activity Relationship
• ISSN: 0021-924X; 1756-2651
• DOI: 10.1093/oxfordjournals.jbchem.a134152

The transfer of phenylalanine from gramicidin S synthetase 1 (GS 1) to gramicidin S synthetase 2 (GS 2) was studied by the use of combinations of wild-type GS 1 with various GS 2s from a wild strain and gramicidin S non-producing mutant strains of Bacillus brevis Nagano. The combinations of mutant GS 2s lacking 4′-phosphopantetheine (from BI-4, C-3, E-1, and E-2) did not transfer D-phenylalanine from GS 1, although they could activate all the constituent amino acids. Other mutant GS 2s containing 4′-phosphopantetheine, except GS 2 from BII-3 (proline-activation lacking) accepted D-phenylalanine from intact GS 1 To ascertain more directly whether 4′-phosphopantetheine is involved in the transfer of D-phenylalanine from OS 1 to GS 2, pepsin digests of GS 2 that accepted [14C]phenylalanine were analyzed by Sephadex G-50 column chromatography and thin-layer chromatography (TLC). Radioactivity of [14C]phenylalanine was always associated with a peptide containing 4′-phosphopantetheine. Furthermore, the position of radioactivity was distinct from the position of 4′-phosphopantetheine on TLC after alkaline treatment or performic acid oxidation of the digests.