Fig 1.
Structure of cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates.
(A) Cinnamoyl anthranilates. Tranilast: R1 = R2 = R3 = R6 = H, R4 = R5 = OMe. (B) Dihydrocinnamoyl anthranilates. DHavnD: R1 = R2 = R3 = R4 = R6 = H, R5 = OH. (C) Benzoyl anthranilates. Dianthramide B from D. caryophyllus: R1-6 = H.
Fig 2.
Strategy used for the biological synthesis of cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates.
Diagram of the reactions catalyzed by 4CL5 and HCBT in the yeast strain engineered for the production of various cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates upon feeding with cinnamates, dihydrocinnamates, or benzoates (donors); and with anthranilates (acceptors). HSCoA, Coenzyme A.
Table 1.
Structural characteristics of the cinnamoyl anthranilates (general structure shown in Fig 1A) produced in yeast and their identification based on dominant ion masses in ESI-MS spectra.
Values were obtained from single feeding experiments for each combination of precursors.
Fig 3.
Detection of N-(4’-hydroxydihydrocinnamoyl)-anthranilate (DHavnD) from the recombinant yeast culture medium.
Representative ESI-MS spectra were obtained after LC-TOF MS analysis of (A) the culture medium of recombinant yeast incubated with anthranilate and 4-hydroxydihydrocinnamate, and (B) a DHavnD standard solution.
Table 2.
Structural characteristics of the dihydrocinnamoyl anthranilates (general structure shown in Fig 1B) produced in yeast and their identification based on dominant ion masses in ESI-MS spectra.
Values were obtained from single feeding experiments for each combination of precursors.
Fig 4.
Detection of N-(benzoyl)-anthranilate (dianthramide B) from the recombinant yeast culture medium.
Representative ESI-MS spectra were obtained after LC-TOF MS analysis of (A) the culture medium of recombinant yeast incubated with anthranilate and benzoic acid, and (B) a dianthramide B standard solution.
Table 3.
Structural characteristics of the benzoyl anthranilates (general structure shown in Fig 1C) produced in yeast and their identification based on dominant ion masses in ESI-MS spectra.
Values were obtained from single feeding experiments for each combination of precursors.