Aspergillus oryzae for producing feruloyl esterase and application thereof
A technology of ferulic acid esterase and Aspergillus oryzae, applied in the biological field, can solve problems such as unfavorable sustainable development, and achieve the effects of easy cultivation and preservation, strong growth ability and rapid reproduction
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Embodiment 1
[0038] Preliminary screening of Aspergillus oryzae, high-yielding strains were screened by the Oxford cup transparent circle screening experiment.
[0039] Soil samples were collected from the soil with a lot of saprophytic plants around South China University of Technology, and 10g of the sample was weighed and put into a 300mL triangular flask filled with 90mL of sterile saline containing glass beads and shaken for 20 minutes to make a 10-fold incremental serial dilution sample Homogenous solution 1:10, 1:100, 1:1000, and 1:10000, take 1.0mL each of the diluted sample homogeneous solution 1:100, 1:1000, and 1:10000 and add it to a sterile plate; add 15mL PDA for culture Mix the medium evenly, let it stand for 1min, and culture it upside down at 28°C for 72h, transfer the single colony on the medium to the slant PDA medium and culture it at 28°C for 72h, and then transfer to the primary screening medium containing ethyl ferulic acid 28 Cultivate at ℃ for 60 hours, screen the ...
Embodiment 2
[0042] Microbiological characteristics and identification of strains
[0043] Aspergillus oryzae ( Aspergillus oryzae ) rice 3 grows slowly on Cha's culture medium, with a diameter of 50mm after 7 days of culture; there are radial irregular grooves, velvet-like texture and flocculent in the center, many conidia structures, no exudate, and the reverse side is Pale pink to light brown. The colony grows fast on malt juice agar, and after 7 days of culture, it covers the entire dish, and the texture is velvety to thick flocculent; the yellow-green color is close to yellow olive, and then becomes light brown olive, and the reverse side is colorless. The colony was cultured on Saccharomyces chagheri paste agar for 7 days, with a diameter of 70mm, radial grooves, velvety or flocculent texture; many conidia structures, light yellowish brown.
[0044] Under the microscope, it can be observed that the conidia heads are spheroid and then radial. The conidiophores occur in the matrix or...
Embodiment 3
[0047] Determination of the Release Amount of Bound Ferulic Acid by Solid State Fermentation
[0048] According to the formula of seed culture medium, weigh 10g of bran and add 10g of water and mix it into a 500mL triangular flask. After the seed culture medium is sterilized, inoculate Aspergillus oryzae ( Aspergillus oryzae ) Mi 3 slant strain, the inoculation amount was 0.5%, and after culturing at 28°C for 3 days, the seeds of Aspergillus oryzae were obtained;
[0049] According to the formula of the fermentation medium, add 200g of bran to 200g of water, mix and sterilize, put it on a porcelain plate to cool, control the thickness of wheat bran within 2.5cm, and the inoculation amount of Aspergillus oryzae seeds is 0.5% (the bacterial content is 1~2×10 9 Individual / g), culture temperature is 28 ℃, carry out ventilated culture, fermentation time is 48h. Solid-state fermented bran was leached in a water bath at 40°C for 60 min at the ratio of 15 g to 90 mL of water, centrif...
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