Preparation and application of a strain of Chaetomium globosa and its metabolite aflatoxin
A technology of aspergillus flavus and Chaetomium globosa, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve the problems of unreported antioxidant activity, etc., and achieve the effect of strong antioxidant activity
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Embodiment 1
[0018] Example 1. Isolation and purification of endophytic strain CDW7:
[0019] Collect fresh and healthy ginkgo leaves, wash and soak in 75% alcohol for 30 seconds, rinse with sterile water for 3 times, then soak in 3-5% NaClO solution for 3-5 minutes, and rinse with sterile water for 3-5 times. Under sterile conditions, cut the surface-sterilized ginkgo leaves into small pieces of 0.5×0.5cm, and place them on PDA for culture at a constant temperature of 25°C. After the bacteria grow out, pick the mycelium at the edge of the colony for purification and culture Until the colonies have the same shape, they are stored on a PDA slant to obtain endophytic bacteria of Ginkgo biloba. One of the strains is numbered CDW7, which is now preserved in the China General Microorganism Culture Collection Center (CGMCC for short), with a preservation date of October 15, 2012 and a preservation number of CGMCC No.6658.
Embodiment 2
[0020] Embodiment 2. Identification of endophytic strain CDW7:
[0021] The endophytic fungus CDW7 is on the PDA plane medium, and the initial stage of the colony is white (see attached figure 1 ), fades to dark cyan (see attached figure 2 ), brown on the back, with irregular edges. Ascothecia are scattered or grouped, ovate or nearly spherical, size (418-610) μm×(222-277) μm (see attached image 3 ); Ascus shell wall is composed of dark brown interlaced filamentous tissue, apical hairs without septa, brown, zigzag or helically curled, with fine spines on the surface, base diameter is 3.1 μm; ascospores are brown, subspherical or lemon-shaped, Size (7.5-10.2) μm × (6.5-7.5) μm, with a terminal bud hole (see attached Figure 4 ). The above morphological characteristics of CDW7 are consistent with the morphological characteristics of Chaetomium sp. fungi.
[0022] The mycelia of the activated CDW7 strain were picked into the PDB medium, cultured at 25°C for 5 days, and the...
Embodiment 3
[0024] Embodiment three. The preparation of compound aflatoxin (flavipin):
[0025] (1) Fermentation culture of CDW7: inoculate the strain of Chaetomium globosa CDW7 stored at 4°C on a PDA plate, culture it in a constant temperature incubator at 25°C for 5 days, and take 12 bacteria cakes with a diameter of 0.5cm on the plate The block was put into a sterilized 250mL Erlenmeyer flask filled with 100mL PDB medium, and cultured at 25°C and 150rpm for 14 days.
[0026] (2) Obtaining of the crude extract of the fermented product: take the fermented liquid obtained in the above step (1), separate the thalline and the fermented liquid with gauze, and obtain 2 L of the total fermented filtrate, and extract the fermented filtrate with dichloromethane solvent for 3-5 times, the combined extracts were concentrated under reduced pressure to obtain the fermented crude extract F1 (2g).
[0027] (3) Separation and purification of the compound: the crude extract of the fermented product obt...
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