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2 protocols using 2 amino 3 methoxybenzoic acid

1

Synthesis of Dihydro-coumaroyl and Coumaroyl Anthranilate Amides

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The chemical standard for methyl 3-hydroxy-2-(3-(4-hydroxyphenyl)propanamido)benzoate [dihydro-coumaroyl anthranilate amide (2)] and (E)-methyl 3-hydroxy-2-(3-(4-hydroxyphenyl)acrylamido)benzoate [coumaroyl anthranilate amide (3)] was purchased from Toronto Research Chemicals (Canada). Methyl 2-amino-3-hydroxybenzoate [3-HAA methyl ester (1)] was purchased from Apollo Scientific (UK). p-Coumaric acid, malonyl-CoA, 3-HAA, methyl 3-hydroxybenzoate, 2-amino-3-methoxybenzoic acid, 2-amino-4-methoxybenzoic acid, 2-amino-5-methoxybenzoic acid, 2-amino-5-hydroxybenzoic acid, and 2-anthranilic acid were purchased from Sigma-Aldrich (St. Louis, MO). Naringenin chalcone was purchased from Biosynth Carbosynth (USA). Naringenin was purchased from MedChemExpress (USA). p-coumaroyl-CoA standard was purchased from PlantMetaChem (Germany).
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2

Optimization of X. eapokensis Growth

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X. eapokensis was grown in liquid SF-900 II SFM (Thermo Fisher Scientific) in presence of 2% Amberlite XAD-16 (Sigmal Aldrich) or SF-900 supplemented with 1 mM of 2-amino-3-chlorobenzoic acid, 2-amino-5-chlorobenzoic acid, 2-amino-3-methylbenzoic acid, 2-amino-5-methylbenzoic acid, 2-amino-3-methoxybenzoic acid, 2-amino-5-methoxybenzoic acid, anthranilic acid, 3-hydroxy anthranilic acid, 3-methyl proline, 3-benzyl proline, or 4-hydroxy proline, pipecolic acid, and 2 mM of 5-fluoro indole, 5-phenyl indole, or 5-methyl indole and 3 mM of 7-azaindole (all chemicals were purchased from Sigma Aldrich). The cultures were cultivated at 30°C for three days with a starting OD600 of 0.1 and cultures were extracted as described below.
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