Bacterial strain for producing citric acid and method for preparing citric acid by fermenting same
A technology of citric acid and strain, applied in the field of fermentation engineering, can solve the problems of high cost, difficult removal of pigment substances, and unsatisfactory results, and achieves the effects of reducing environmental pollution and reducing production costs.
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Embodiment 1
[0031] Example 1 Screening of bacterial strains
[0032] (1) Separation and purification of strains: Aspergillus niger (Aspergillus niger, preservation number CGMCC NO.6465) was used as the starting strain, and it was diluted and separated under aseptic conditions, spread on the plate medium, and stored at 36°C Incubate at ±1°C for 40 hours;
[0033] Described plate medium is PDA potato medium, wherein 12% of potato juice, 3% of glucose, 2% of agar are prepared using neutralized waste water instead of tap water;
[0034] (2) Expansion culture of strains: After the single colonies on the plate have been cultured, select several groups of single colonies that have grown from the cultured plates and inoculate them on the slant medium of the test tube, and culture at 36°C±1°C for 5 days;
[0035] Described plate medium is PDA potato medium, wherein 13% of potato juice, 2% of glucose, 2% of agar are prepared using neutralized waste water instead of tap water;
[0036] (3) Strain ...
experiment example 1
[0040] Experimental Example 1 Investigation of Acid Production Levels in Different Shake Flask Fermentation Cultures
[0041] 1) Effects of shake flask fermentation media prepared with different solutions on the acid production level of the strain
[0042] Aspergillus niger (Aspergillus niger, preservation number CGMCC NO.6465) and the strain obtained in Example 1 were subjected to shake-flask fermentation acid production tests in shake-flask fermentation medium 1 and 2, respectively, and the specific results are shown in Table 1.
[0043] Among them, shake flask fermentation conditions: 36°C±1°C, shaker speed 320rpm / min, culture for 96h;
[0044]The shake flask fermentation medium 1: the corn flour raw material is adjusted to a powder slurry ratio of 20%, α-amylase is added, liquefied, filtered, and the obtained filtrate is used as a base material, and it is prepared by using tap water, wherein the pH before sterilization is 6 .02, pH5.59 after sterilization, total sugar 0.1...
Embodiment 2
[0054] Embodiment 2 The method for producing citric acid in a 50L fermenter by the bacterial strain obtained in Example 1
[0055] (1) Expansion culture: take the strain obtained in Example 1 as the starting strain, insert the slant of the subculture test tube for the first expansion culture, the culture condition is 36°C±1°C, and the culture time is 5d; then the cultivated slant Put the spores into the bran yeast bottle for the second expansion culture, the culture condition is 36°C±1°C, and the culture time is 7 days;
[0056] (2) Preparation of spore suspension: wash the spores cultivated in step (1) to prepare a suspension, and the number of spores per gram of bran koji is 3×10 10 , pour the spore suspension steel bottle under aseptic conditions for standby;
[0057] (3) Seed tank cultivation: Put the spore suspension obtained in step (2) into a citric acid seed tank for cultivation. The cultivation conditions are: 36°C±1°C, ventilation ratio 1:1, tank pressure 0.5MPa, T...
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