Chemical synthesis and characterization of the sweet protein mabinlin II.
The sweet protein mabinlin II isolated from the seeds of Capparis masaikai consists of the A chain with 33 amino acid residues and the B chain composed of 72 residues. The B chain contains two intramolecular disulfide bonds and is connected to the A chain through two intermolecular disulfide bridges. The A chain was synthesized by the stepwise fluoren-9-ylmethoxycarbonyl (Fmoc) solid-phase method in a yield of 5.9%, while the B chain was synthesized by a combination of the stepwise Fmoc solid-phase method and fragment condensation in a yield of 6.0%. Disulfide formation and combination of the A and B chains followed by purification by ion-exchange high-performance liquid chromatography (HPLC) gave mabinlin II in a yield of 47.4%. The characterization of the synthetic mabinlin II by HPLC, electrospray ionization mass spectrometry, amino acid analysis, and disulfide bond determination fully supported the expected structure. A 0.1% solution of the synthetic mabinlin II had an astringent-sweet taste.
Central Research Laboratories, Ajinomoto Co., Inc., Kawasaki, Japan.
M
M
Kohmura
Y
Y
Ariyoshi
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D003545:Cysteine / Q000032:analysis; D004220:Disulfides / Q000032:analysis; D005449:Fluorenes / Q000032:analysis; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000138:chemical synthesis; D010940:Plant Proteins / Q000032:analysis / Q000138:chemical synthesis*; D013549:Sweetening Agents / Q000032:analysis / Q000138:chemical synthesis*; D013649:Taste
D000596:Amino Acids; D004220:Disulfides; D005449:Fluorenes; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D010446:Peptide Fragments; D010940:Plant Proteins; D013549:Sweetening Agents; C083584:mabinlin protein, Capparis masaikai; D003545:Cysteine