Hygrovetine → (2S,3'R,3aR,4R,4'S,5'R,6R,6'R,7S,7aS)-4-[(1S,2R,3S,5R,6S)-3-amino-2,6-dihydroxy-5-(methylamino)cyclohexyl]oxy-6'-[(1R)-1-amino-2-hydroxyethyl]-6-(hydroxymethyl)spiro[4,6,7,7a-tetrahydro-3aH-[1,3]dioxolo[4,5-c]pyran-2,2'-oxane]-3',4',5',7-tetrol

After the incubation in Example 2, each culture was diluted with water to an OD600 of 0.5 (cell count 107 cells/ml) and further diluted by a factor of 10 (in the far left column marked with 10−1 in FIG. 3), and 7 μl of each dilution was spotted onto an YES medium (agar plate) and incubated at 30° C. (FIG. 3(a)) and 37° C. (FIG. 3 (c)) for 3 days. Similarly, each dilution was incubated on YES medium (5 mL) containing 20 μg/l hygromycin B (0.05 mass %) at 30° C. overnight (FIG. 3 (b)). If a rapid growth was observed in a short time, the dilutions were cultured under similar conditions after further serially diluted by a factor of 10. In FIG. 3 (a) to (c), cultures of serial 10-fold dilutions from OD600 of 0.5 (10−1, 10−2, 10−3 and 10−4) are shown.

1. customincubated at 30° C.
2. customfor 3 days

ORD-Eintrag

ord-6c7671ce092b42468fa0cd9a73ff70d6

Quelldatensatz

uspto-grants-2012_11

DOI

10.6084/m9.figshare.5104873.v1

Patentnummer

US08318454B2