A kind of phthalate ester and its preparation method, bacterial strain and application
A technology of phthalates and seeds, applied in the field of symbiotic fungus - anthracnose, can solve the problems of unstable target products and difficult waste liquid treatment, and achieve the effects of short cycle, guaranteed source and low cost
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Embodiment 1
[0025] Embodiment 1: the isolation and identification of ball anthracnose bacterium (Colletotrichum coccodes) ZZP-L1
[0026] 1. Isolation of strain ZZP-L1
[0027] The final concentration of the slant medium (ie PDA medium) consists of: sucrose 20g / L, potato 200g / L, agar 20g / L, the solvent is distilled water, and the natural pH value.
[0028] The final concentration of the seed medium (ie PDB medium) consists of: 200 g / L of potatoes, 20 g / L of sucrose, distilled water as a solvent, and natural pH.
[0029] Strain ZZP-L1 was first isolated from the coastal plant Rumexobtusifolius Linn. by Mr. Zhang Huawei from Zhejiang University of Technology using common symbiotic bacteria isolation techniques (for details, refer to Bangladesh Journal of Pharmacology, 2012, 7:120-123), and was inoculated in Inclined media.
[0030] 2. Identification of strain ZZP-L1
[0031] (1) Morphological characteristics: strain ZZP-L1 was inoculated on PDA medium, activated at 28°C for 3-4 days, and...
Embodiment 2
[0035] Embodiment 2: the preparation of globular anthracnose bacteria ZZP-L1 fermentation medium
[0036] (1) Slant culture: inoculate the slant culture medium with Bacillus anthracis ZZP-L1, and culture in a 28°C incubator for 3-4 days to obtain a slant surface; the final concentration composition of the slant medium is the same as in Example 1.
[0037] (2) Seed culture: pick an inoculation ring from the slant of the thalline and inoculate it into the seed medium, and culture it on a shaker at 200 rpm and 28°C for 3 days to obtain the seed liquid; the final concentration of the seed medium is composed with the same implementation example 1.
[0038] (3) Fermentation culture: the seed liquid obtained in step (2) is inoculated to the fermentation medium with an inoculum size of 10% volume concentration, and cultured on a shaker at 200 rpm and 28° C. for 10 days to obtain a fermentation medium, a fermentation medium The final concentration is the same as that of the seed mediu...
Embodiment 3
[0039] Embodiment 3: Extraction and separation, identification of two kinds of phthalates
[0040] 1. Extraction and separation of two phthalates
[0041] After the fermentation broth obtained in Example 2 was filtered, the filtrate was extracted twice at room temperature with ethyl acetate (volume ratio 1:1), and the upper organic phase was taken and concentrated in vacuum at low temperature (35°C, 0.01MPa) until no liquid flowed out. Dry to obtain brown crude extract F; the methanol solution of crude extract F is separated by reversed-phase high-performance liquid chromatography (RP-HPLC), and the peaks at the 6th and 14th minutes are collected respectively, and the mobile extract F is removed under low-temperature vacuum (35°C, 0.01MPa). Phase to obtain compound (I), compound (II).
[0042] Liquid phase instrument: UV-VIS, detector: Shimadzu SPD-10AV; HPLC infusion pump: Shimadzu LC-10AD; chromatographic conditions: C 18 The chromatographic column is 250×9.4mm, the flow r...
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