Streptomyces albulus (GS-114) and method of the same to prepare epsilon-polylysine
A technology of Streptomyces parvus and microbial strains, applied in the field of Streptomyces and its preparation of ε-polylysine, can solve the problems of not being suitable for large-scale application in breeding work, general screening effect, and high price, and achieve The effect of excellent passage stability
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Embodiment 1
[0049] Embodiment 1: the breeding of Streptomyces parvus GS-114
[0050] Bethner solid medium (g / 100mL): glucose 2, yeast extract 0.2, peptone 0.4, agar 2, pH 7.5, sterilized at 115°C for 20 minutes.
[0051] Antibiotic resistance medium (g / 100mL): glucose 2, yeast extract 0.2, peptone 0.4, agar 2, pH 7.5, sterilized at 115°C for 20 minutes. When the temperature of the culture medium dropped to 60°C, the stock solutions of streptomycin, gentamicin and rifamycin prepared in advance and sterilized by filtration through a 0.22 μm membrane were added thereto.
[0052] Specific steps are as follows:
[0053] (1) Construction of mutation library: 2% (v / v) DES was used to test the spores of Streptomyces albicans M-Z18 (the deposit number is CCTCC NO: M 2019589, and the strain is recorded in the patent application with application number 201911020454.4) The suspension was treated for 30min (28°C water bath), and NaS was added 2 o 3 Terminate the reaction, dilute and smear on a Bet...
Embodiment 2
[0055] Example 2: Morphological characteristics of the starting bacterium M-Z18 and the high-yield bacterium GS-114
[0056] The morphology of the strains was observed with naked eyes and microscopic analysis.
[0057] Depend on figure 1 It can be seen that there are obvious differences in morphology between the two strains. On the peptone yeast extract medium, the colony of the starting strain M-Z18 was larger, the aerial hyphae grew well, and the colony and its back were yellow or tan. In YP liquid medium, the mycelium of M-Z18 intertwined to form larger solid balls.
[0058] The colony of strain GS-114 is small. On the peptone yeast extract medium, the aerial hyphae grow well, and the spores form gray-green chains, which are oval under the microscope; the colonies and their backs are yellow-green or yellow-brown. In the YP liquid medium, the hyphae of GS-114 were loose and the balls were small.
Embodiment 3
[0059] Example 3: The difference in the shake flask output of high-yielding bacteria GS-114 and starting bacteria M-Z18 under different medium conditions
[0060] M3G seed medium (by g / 100mL): glucose 5, (NH 4 ) 2 SO 4 1, Na 2 HPO 4 0.14, KH 2 PO 4 0.1, MgSO 7 ·H 2 O 0.025, ZnSO 7 ·H 2 O 0.005, FeSO 7 ·H 2 O 0.001, yeast powder 0.5, pH 6.8, sterilized at 115°C for 20min.
[0061] RSM liquid fermentation medium (by g / 100mL): glucose 6, (NH 4 ) 2 SO 4 0.5, beef extract 1, KH 2 PO 4 0.4, MgSO 4 0.08, FeSO 4 0.005, sterilized at 115°C for 20 minutes, fermented at natural pH.
[0062] YP liquid fermentation medium (by g / 100mL): glucose 3, (NH 4 ) 2 SO 4 0.5, yeast powder 0.8, KH 2 PO 4 0.4, MgSO 4 0.08, FeSO 4 0.005, sterilized at 115°C for 20 minutes, fermented at natural pH.
[0063] The purified GS-114 spores were inoculated into the medium filled with M3G, and cultured in shake flasks at 30°C×200rpm for 24h to prepare seed liquid. The seed so...
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