Aspergillus niger capable of highly yielding compound enzyme for feed and application thereof
A compound enzyme for feed, Aspergillus niger technology, applied in the direction of enzymes, lyases, hydrolytic enzymes, etc., can solve the problems of popularization and application obstacles, difficulties in growth and synthesis, and affecting the effect of use, so as to achieve good economic benefits and feed The effect of compound enzyme with less impurities and improved digestibility
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Embodiment 1
[0028] Isolation, purification and identification of the Aspergillus niger DM-18 original strain of high-yield feed compound enzyme of embodiment 1
[0029](1) Strain isolation: Collect 10g of humus soil and rice straw mixed soil sample from Yangyou Township, Jin City, Hunan Province, add it to a small triangular flask containing 100mL of cooled sterile water with glass beads, shake at 200r / min, and shake Mix evenly for 30 minutes, then place in an 80°C water bath for 10 minutes, shake the Erlenmeyer flask constantly to mix the soil sample, let it stand for 5 minutes, draw 100 μL of the supernatant, and gradually dilute to 10 -1 ~10 -9 Concentration, select 10 -3 , 10 -4 , 10 -5 , 10 -6 Draw 0.2ml of the diluted solution and spread it on the PDA plate. After 4-5 days of constant temperature cultivation at 28°C, the colony will cover the entire plate.
[0030] (2) Strain purification: According to the morphological characteristics of the primary colony, a small amount of m...
Embodiment 2
[0037] The screening method of the bacterial strain of embodiment 2 high-yield compound enzymes for feed, comprises the following steps
[0038] 1) Slant culture: the isolated and purified Aspergillus niger original strain HYX0022 in Example 1 was streaked and inoculated on the slant medium, and cultured at 28-33° C. for 3-4 days until the mycelia matured and produced spores.
[0039] The composition of the slant medium is as follows: 200g of potatoes (boiled juice), 20g of glucose, 20g of agar, 1000mL of distilled water, pH value 5.8, sterilized at 121°C for 20min;
[0040] 2) Preparation of spore suspension: when Aspergillus niger produces spores, add 5 mL of sterile water to the inclined surface of the test tube, scrape off the spores, pour them into a sterilized triangular flask and add Tween-80 with 0.5-2% spore mass, Put the triangular flask in a shaker and vibrate for 1.5h to activate the spores; then centrifuge at 3500r / min for 15min, wash once with pH7.2 phosphate buf...
Embodiment 3
[0066] Example 3 The culture medium optimization of the starting strain for high-yield feed compound enzyme screening
[0067] (1) Strains
[0068] The starting strain is the DM-18 strain with the best screening results. After 96 hours of fermentation, the crude enzyme liquid of the fermentation broth contains a variety of enzymes, among which the protease activity is 6300U / ml, the mannanase activity is 1350U / ml, and the α -Galactase activity 1100U / ml, pectinase activity 900U / ml.
[0069] (2) Culture medium optimization
[0070] Optimizing the slant culture medium, plate culture medium, seed culture medium and fermentation medium in the process of strain screening, and fermenting for 96 hours under the same fermentation conditions before optimization. After testing, the crude enzyme liquid of the fermentation broth contains a variety of enzymes, Among them, protease activity 600U / ml, mannanase activity 1500U / ml, α-galactosidase activity 1200U / ml, pectinase activity
[0071]...
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