Apergillus niger strain and application thereof
A technology of Aspergillus niger and strains, applied to the Aspergillus niger ML-0016 strain and its application field, can solve the problems of less research and achieve the effect of mild reaction conditions
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Embodiment 1
[0024] Breeding of Aspergillus niger ML-0016 Strain
[0025]The starting strain was screened from the soybean paste sample, and then cultured on an inclined plane (28°C, 2-3 days) to obtain a spore suspension; after treatment with NTG for 2, 4, 6, 8, and 10 minutes, the mutagenesis was terminated, and the treated spores were suspended Incubate for 4 to 5 days at 28°C after diluted with liquid and spread on the plate. The colony on the plate was picked out, streaked to isolate the pure species, and then the shake flask was used for preliminary screening (28° C., 50 h). Screen the strains and select one as the starting strain for further mutagenesis, prepare the spore suspension, and use 60 Coγray irradiation doses were 0 rad, 30,000 rad, 50,000 rad, 75,000 rad, 100,000 rad, and 150,000 rad, respectively. The irradiated spore suspension was plated and cultured (28°C, 4-5d), the colonies on the plate were picked out, and the pure species was separated by streaking, and then the...
Embodiment 2
[0030] Medium formula (weight / volume percentage, the same below): defatted soybean meal 4%, bran 2%, peptone 0.1%, (NH 4 ) 2 SO 4 0.1%, KH 2 PO 4 0.1%, Sodium Acetate 0.1%, Ascorbic Acid 0.1%, MgSO 4 0.1%, NaH 2 PO 4 2H 2 O: 0.4%, salicin 0.05%, pH 6.0, prepared with tap water, sterilized at 121°C for 20 minutes for later use, in this example, the defatted soybean meal can use the soybean meal after industrial oil extraction, and its consumption can be controlled at 0.5 %~12% range.
[0031] Take 100mL of the above-mentioned culture medium, divide it equally into two 250mL Erlenmeyer flasks, and sterilize. Inoculate the cultured cells of the slant strain of Aspergillus niger ML-0016 strain with the preservation number CCTCC M 206034 at a shaker speed of 150r / min, and culture on a shaker at 28°C for 24 hours as a seed solution for later use.
[0032] Take 2L of the above culture medium, divide it into 40 500mL shake flasks, and sterilize. The seed liquid was insert...
Embodiment 3
[0034] 0.02M, pH 5.0 HAc-NaAc buffer solution is used to formulate soybean isoflavones into a substrate solution with a concentration of 1-10 mg / ml, and an appropriate amount of ethanol is added as a substrate auxiliary agent. Take 50 ml of the crude enzyme solution prepared in Example 1, mix the crude enzyme solution and the substrate solution (substrate concentration is 0.5% to 15.0%) in a ratio of 1:1, and carry out enzymolysis at 20°C to 50°C After 1-10 hours of reaction, 100 ml of enzymolysis solution was obtained.
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