Reference

Fermentative production of FAD

Fermentative production of FAD. (Kyowa Hakko Kogyo Co., Ltd., Japan). Jpn. Kokai Tokkyo Koho JP 59132898 A2 31 Jul 1984 Showa, 4 pp. (Japanese). (Japan). CODEN: JKXXAF. CLASS: IC: C12P019-36. ICI: C12P019-36, C12R001-15; C12P019-36, C12R001-13. APPLICATION: JP 83-4719 14 Jan 1983. DOCUMENT TYPE: Patent CA Section: 16 (Fermentation and Bioindustrial Chemistry) FAD [146-14-5] was prepd. in a 2-step process with Corynebacterium or Brevibacterium, capable of forming ATP [56-65-5] from a ATP precursor, i.e., adenine [73-24-5], adenosine [58-61-7], 51-AMP [61-19-8] and ADP [58-64-0], in the presence of phosphate and energy donors, and also producing FAD from ATP and FMN [146-17-8]. Thus, B. ammoniagenes ATCC 21170 was inoculated into 800 mL medium (pH 7.2) contg. glucose 150, urea 5, yeast ext. 5, meat ext. 5, KH2PO4 10, K2HPO4 10, MgSO4.7H2O 10, CaCl2.2H2O 0.1 g and biotin 30 mg/L and incubated at 32° for 24 h with stirring and aeration, maintaining the pH at 6.8 with NH3. The broth was centrifuged and the cells suspended in 500 mL soln. contg. glucose 50 adenine 4, NaH2PO4 15, xylene 0.01, and surfactant 6 g/L and incubated at 32° for 10 h with stirring and aeration: the pH and the H3PO4 concn. were kept at 7.4 and 10 g/L (as NaH2PO4), resp. Then, 3 g Na-FMN/L was added and incubated at 42° for 12 h. The resulting mixt. (1 L) was heat-treated, filtered and the supernatant made pH 3.5, adsorbed to active C and eluted with 50% EtOH. The eluate was concd., chromatographed on Dowex 1 ′ 2 and active C and FAD was pptd. as a Ba salt, which was mixed with Na2SO4 to give 1.35 g FAD-Na2 [84366-81-4] (as FAD).