Aspergillus niger WZ001 capable of producing naringinase and cirmtimase simultaneously and its application
A technology of Aspergillus niger and hesperidinase, applied in the direction of bacteria, hydrolase, and microorganism-based methods, can solve the problem of lack of high-yielding naringinase and hesperidinase strains, unoptimized fermentation conditions, and enzyme production minor issues
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Embodiment 1
[0020] After the Aspergillus niger WZ001 strain was activated by Chase's medium for 5 days, 30ml of distilled water was taken to make a spore suspension.
[0021] Quality composition of liquid fermentation medium: bran 5%, bean cake powder 4%, bean dregs 10%, the balance is water, pH is natural, 0.1Mpa steam pressure sterilization for 20min, after cooling, add black koji to each bottle at a volume ratio of 2% Mold suspension, cultured for 5-7 days at a fermentation temperature of 30-37°C and a rotational speed of 150-200r / min, and filtered to obtain a crude enzyme solution, in which the activity of hesperidinase was measured to be 14104.9U / g fermentation raw material, pomelo The enzymatic activity of glucosidase was 14799.6U / g fermentation raw material.
Embodiment 2
[0023] After the Aspergillus niger WZ001 strain was activated by Chase's medium for 5 days, inoculate a loop into the liquid seed medium (4% bran, 3% bean cake powder, 8% bean dregs, and the balance was water) and cultivate for 36 hours to obtain the seed liquid .
[0024] The solid-state fermentation medium is bran 100g, bean dregs 10g, bean cake powder 20g, add 100ml of water, pack in a 500ml Erlenmeyer flask, the thickness of the filling is 2.5-3.0cm, 0.1Mpa steam pressure sterilization for 30min, after cooling, each The bottle was inserted into the Aspergillus niger seed solution at a volume ratio of 2%, cultivated at 32°C for 5-7 days, soaked in water for 4 hours, and filtered to obtain a crude enzyme solution, in which the enzyme activity of hesperidinase was measured to be 6378.5U / g Fermented raw materials, the enzyme activity of naringinase is 6601.7U / g fermented raw materials.
Embodiment 3
[0026] Add 200 mL of ethanol to 100 mL of the crude enzyme solution obtained by the method in Example 2, let stand for 20 min, centrifuge, and dry to obtain mixed enzyme powder of naringinase and hesperidinase. The recovery rate of hesperidinase was 75.2%, and that of naringinase was 68.7%.
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