Method for producing feruloyl esterase by solid fermentation
A technology of ferulic acid esterase and solid fermentation, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of low enzyme production and enzyme activity, achieve high enzyme activity, reduce pollution, and reduce production cost effect
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[0034] (1) Preparation of spore suspension:
[0035] Inoculate the Aspergillus niger with the preservation number CGMCC No.6152 on the slant medium according to the streaking method, and cultivate it at a constant temperature of 28°C for 7 days. At this time, the strain has matured. for 10 6 ~10 7 spore suspension per ml, ready for use.
[0036] (2) Seed preparation:
[0037] Firstly, the liquid seed medium was sterilized at 121°C for 20 minutes, and then cooled; then the spore suspension was inserted into the above-mentioned cooled liquid seed medium according to the volume ratio (V / V) of 5% inoculum, and the medium was incubated at 28 ° C, cultivated on a shaker with a rotational speed of 200 rpm for 18 hours to obtain liquid seeds.
[0038] (3) Ferulic esterase fermentation
[0039] Preparation of medium for fermentation;
[0040] Cover the mouth of the Erlenmeyer flask containing the medium for fermentation with 8 layers of gauze, sterilize the fermentation medium at...
Embodiment 1
[0048] Aspergillus niger CGMCC No.6152 obtained by separation and purification was used as the strain, and the spore suspension was collected. After the seeds were prepared, the seeds were inoculated into the fermentation medium for fermentation and cultivation. The solid matter after the step measures the enzyme activity.
[0049] The preparation process of the fermentation medium is as follows: in a 250ml Erlenmeyer flask, 5g of carbon sources are respectively 2.8g of bran and 2.2g of corncob powder, 0.4g of peptone, and 0.12g of KH 2 PO 4 , 0.06gNa 2 HPO 4 ·7H 2 O, 0.001g NaCl, 0.002g MgSO 4 7H2O, 0.0005g CaCl 2 Then add 15ml of water to the bottle, the pH is natural.
[0050] After determination, the enzyme activity of ferulic esterase was 22.04U / g dry koji.
Embodiment 2
[0052] Aspergillus niger CGMCC No.6152 obtained by separation and purification was used as the strain, and the spore suspension was collected. After the seeds were prepared, the seeds were inoculated into the fermentation medium for fermentation and cultivation. The solid matter after the step measures the enzyme activity.
[0053] The preparation process of the fermentation medium is as follows: in a 250ml Erlenmeyer flask, 5g of carbon sources are respectively 2.5g of soybean meal and 2.5g of corncob powder, 0.2g of peptone, 0.08g of KH 2 PO 4 , 0.06gNa 2 HPO 4 ·7H 2 O, 0.001g NaCl, 0.002g MgSO 4 ·7H 2 O, 0.0005g CaCl 2 Then add 20ml of water to the bottle to neutralize the pH.
[0054] After determination, the enzyme activity of ferulic esterase was 2.99U / g dry koji.
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