A kind of method utilizing molasses fermentation to produce L-malic acid
A technology of malic acid and molasses, applied in the field of microbial fermentation, can solve the problem that acid production cannot reach large-scale production, etc., and achieve the effects of avoiding the input of steps and the operation of related equipment, increasing the output, and improving the restriction of mass transfer.
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Embodiment 1
[0020] A method of utilizing molasses fermentation to produce L-malic acid, comprising the steps:
[0021] Molasses pretreatment: add twice the mass of purified water to the cane molasses, stir at 1000rpm for 3min, then add activated carbon accounting for 2% of the mass of the molasses, stir at 200rpm for 30min, filter, and collect the filtrate;
[0022] Preparation of fermentation medium: According to 1L medium, add 70g calcium carbonate, 20g glucose, 10g corn steep liquor, 8g urea, 0.1g magnesium sulfate, 0.1g potassium dihydrogen phosphate and 0.1g dipotassium hydrogen phosphate to step 1) In the obtained filtrate, set the volume to 1L and control the pH value to 6.2;
[0023] Fermentation for acid production: according to the volume ratio of Aspergillus oryzae seed liquor: fermentation medium of 1:10, it was transferred to the fermentor for cultivation, the temperature was 33 ° C, and the cultivation was carried out for 48 h, and then calcium carbonate and cetyltrimethylam...
Embodiment 2
[0026] A method of utilizing molasses fermentation to produce L-malic acid, comprising the steps:
[0027] Molasses pretreatment: add twice the mass of purified water to the cane molasses, stir at 1000rpm for 3min, then add activated carbon accounting for 2% of the mass of the molasses, stir at 200rpm for 30min, filter, and collect the filtrate;
[0028] Preparation of fermentation medium: According to 1L medium, add 70g calcium carbonate, 20g glucose, 10g corn steep liquor, 8g urea, 0.1g magnesium sulfate, 0.1g potassium dihydrogen phosphate and 0.1g dipotassium hydrogen phosphate to step 1) In the obtained filtrate, set the volume to 1L and control the pH value to 6.2;
[0029] Fermentation to produce acid: According to the volume ratio of Aspergillus oryzae seed liquor: fermentation medium of 1:10, it was transferred to a fermenter for cultivation, the temperature was 33 ° C, and the cultivation was carried out for 48 hours, and then calcium carbonate and cetyltrimethylammo...
Embodiment 3
[0032] The influence of various factors on the yield of malic acid fermented by Aspergillus oryzae:
[0033] Determination of L-malic acid: using the 2,7-naphthalenediol colorimetric method, take 1.0 mL of the sample solution, add 6.0 mL of analytically pure concentrated sulfuric acid, then add 0.1 mL of 2,7-naphthalenediol solution, and then at 100 Heat in a water bath for 20 min, take it out and cool it to room temperature, perform colorimetric measurement at 385 nm, and use distilled water as a control to calibrate the zero point of the instrument. Use the standard sample to make a standard curve first, take the malic acid content as the abscissa, and the absorption value at 385 nm, i.e. OD385, as the ordinate. Through the OD value of the unknown sample at 385 nm, the corresponding malic acid content can be found on the standard curve. .
[0034] A control group was set, wherein the control group 1: did not increase the temperature and pressure, and the rest were the same ...
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