Strain and method for producing succinic acid by fermenting xylose mother liquor
A technology for producing xylose mother liquor and succinic acid, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems of low yield, complicated process, high raw material cost, etc., and achieve reduction of production cost, production method Simple, effective food security
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Embodiment 1
[0028] Breeding and identification of Actinobacillus succinate
[0029] 1. The method of directed evolution breeding of Actinobacillus succinogenes is as follows: first use 10g / L glycerol to replace the glucose in the seed solution as a carbon source as a directed medium, place it in an anaerobic incubator after inoculation, and keep it at 37°C Cultivate for 12-18 hours for subculture. After 7-10 days of cultivation, the concentration of glycerol increases to 20g / L, and so on. After about 90 days of domestication, the concentration of glycerol in the medium can increase to 80g / L, and a succinic acid-producing actinomycetes can be obtained by breeding. Bacillus PZ, the strain can grow well in the culture concentration of glycerol concentration 80g / L. At the same time, it was found that Actinobacillus succinogenes PZ could grow better in 100g / L xylose mother liquor, and could ferment and produce high concentration of succinic acid.
[0030]2. The 16S rDNA gene identification of...
Embodiment 2
[0033] A method for producing succinic acid by fermenting xylose mother liquor, comprising the following steps:
[0034] Inoculate the single colony of Actinobacillus succinogenes PZ on the plate into the seed medium, and culture it in an anaerobic incubator at 37°C for 16 hours, and the number of bacteria will reach more than 300 million, and then proceed according to the inoculation amount of 5% (V / V) Secondary multiplication and cultivation for 8 hours, when the number of bacteria reaches more than 300 million, liquid seeds are obtained; the liquid seeds are inoculated into a 250mL anaerobic bottle containing a fermentation medium at an inoculation volume of 5% (V / V). 150mL, use pH buffer to adjust the pH of the fermentation broth to maintain at 6.5-7.0, and fill it with N 2 In an environment with a rotating speed of 100r / min and a temperature of 36°C, ferment in batches for 30 hours to obtain succinic acid.
[0035] The composition and concentration of the seed medium are...
Embodiment 3
[0046] A method for producing succinic acid by fermenting xylose mother liquor, comprising the following steps:
[0047] Inoculate the single colony of Actinobacillus succinogenes PZ on the plate into the seed medium, and cultivate it in an ordinary incubator at 38°C for 20 hours, and the number of bacteria reaches more than 300 million. Level multiplication culture 8h, bacterial number reaches more than 300,000,000, promptly obtains liquid seed; By 15% (V / V) inoculum size, liquid seed is inoculated in the 250mL anaerobic bottle that fermentation medium is housed, and liquid capacity is 150mL, use the pH buffer to adjust the pH of the fermentation broth to maintain at 6.5-7.0, 2 In an environment with a rotating speed of 200r / min and a temperature of 38°C, ferment in batches for 72 hours to obtain succinic acid.
[0048] The composition and concentration of the seed medium are: glucose 8g / L, yeast powder 12g / L, corn steep liquor 4g / L, NaHCO 3 4g / L, NaH 2 PO 4 9.6g / L, K 2 ...
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