Penicillium velutinum DZ-9-67 and application thereof in extraction offolium cortex eucommiae total flavonoids
A technology of Penicillium rugosa and Eucommia ulmoides, which is applied in the field of biochemical industry, can solve the problems of high equipment requirements, low yield of total flavonoids, complex semi-bionic extraction process, etc., and achieve the effect of easy dissolution and improved extraction yield
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Embodiment 1
[0021] Example 1: Isolation and screening of fermentation strains
[0022] The microbial strains fermenting Eucommia leaves were isolated and screened according to the following steps:
[0023] (1) About 5 g of eucommia leaf powder was added to a 250-mL Erlenmeyer flask, and then 5 mL of sterile saline was added to moisten it, and incubated at a constant temperature of 28°C for 4 days. The enriched cultures covered with mold were diluted 1×10 with sterile saline -6 , 1×10 -7 , 1×10 -8 After doubling, draw 0.1mL of the diluted solution and spread it on the potato dextrose agar plate medium (PDA), culture at a constant temperature of 28°C for 2 days, pick mold colonies with different colors and shapes and transfer them to fresh PDA plate medium, and place them at 28°C. After culturing at constant temperature at ℃ for 3 days, 14 purely cultured strains were obtained, and the numbers of each strain are shown in Table 1.
[0024] (2) Add 5 mL of sterile saline to the fresh plat...
Embodiment 2
[0036] Embodiment 2: the mutagenesis selection of fermentation strain DZ-9
[0037] Carry out mutagenesis breeding on strain DZ-9, and screen strains with excellent fermentation performance, the specific method is as follows:
[0038] (1) Preparation of spore liquid: After strain DZ-9 was activated and cultured on PDA plate medium at 28°C for 2 days, 5 mL of sterile normal saline was added, stirred with an inoculation loop to suspend the spores, and then transferred to a triangle containing 45 mL of sterile normal saline. In the bottle (with 20–30 glass beads), shake at room temperature for 20 minutes. The spore suspension was filtered to remove mycelium (triangular funnel pad with 2 layers of lens-cleaning paper), counted the spores in the suspension with a hemocytometer under a microscope, and diluted it with sterile saline to adjust the number of spores to 1× 10 8 individual / mL.
[0039] (2) Mutagenesis: under red light illumination, take 2.0mL of the above spore suspens...
Embodiment 3
[0041] Example 3: Classification and Identification of Bacterial Strain DZ-9-67
[0042] Strain DZ-9-67 was inoculated on PDA plate medium and cultured at 28°C for 3 days. The colonies were grayish-white fine short hairs at the beginning, and then gradually turned green to olive green. Produces a large number of powdery conidia. The top of the aerial hyphae produces conidiophores, with 1–4 branches at the top of the stalk, and the terminal cells of each branch divide into clusters of conidia, forming a typical brush-shaped conidia spike; the conidia are spherical or Approximately spherical, 2.5–3.0 μm in diameter, green. See the photos of the colony of Penicillium papaya DZ-9-67 cultured on PDA plate medium at 28°C for 3 days figure 1 .
[0043] The strain DZ-9-67 was handed over to Sangon Bioengineering (Shanghai) Co., Ltd. and the rDNA nucleotide sequence of its ribosomal ITS region was measured as SEQ ID NO.1, which is listed in NCBI (National Center for BiotechnologyInf...
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