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A technology of sialyllactose and lactose, which is applied in the field of bioengineering, can solve the problems that the simulated human milk effect cannot be satisfied, and the difference is large.
Inactive Publication Date: 2011-08-17
朱莉
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However, the composition and content of sialic acid oligosaccharides in these exogenous added substances are quite different from those of human milk, which cannot meet the effect of simulating human milk.
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Embodiment 1
[0023] Example 1 Fermentation
[0024] (1) High-density culture: use Escherichia coli CCTCC NO: M208088 as the strain, add 18L of fermentation medium to a 30L fermenter with an inoculum of 4%, maintain the temperature at 30-37°C throughout the fermentation process, and ventilate at 1- 1.5 vvm, the stirring speed is 600-800 r / min; during the fermentation process, ammonia water is added to control the pH to 6-7, and the fermentation cycle is 32-48 hours; after 2-6 hours of fermentation, feed medium is added until the end of fermentation The addition was stopped half an hour before, and the volume of the added feed medium was equivalent to the volume of the fermentation medium, and was added to the fermentation system at an even speed.
[0026] Feed medium composition g / L: sorbitol 400, ammonium sulfate 50.
[00...
Embodiment 2
[0028] The collection of embodiment 2 sialyllactose
[0029] (1) Sterilization by centrifugation: After the fermentation, the fermented liquid obtained in Example 1 was heated at 80°C for 30 minutes, and then centrifuged at 6000 rpm for 10 minutes to remove bacteria, and supernatant 1 containing sialyllactose and bacteria sludge were obtained.
[0030] (2) Heating and lysing the bacteria: Dilute the bacteria slime obtained in step (1) with water 3 to 5 times, heat in a boiling water bath for 45 minutes, and then centrifuge at 10,000 rpm for 10 minutes to remove cell debris to obtain supernatant 2 containing sialyllactose;
[0031] (3) Ion exchange resin adsorption and desorption: Combine the supernatant 1 obtained in step (1) and the supernatant 2 obtained in step (2), heat to 80-90°C, and heat at constant temperature for 15 minutes; The membrane is subjected to ultrafiltration to collect the filtrate; the filtrate is treated with a molecular weight cut-off of 500Da ultrafiltr...
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Abstract
The invention provides a preparation method of 3'-sialic acid lactose, belonging to the technical field of biological engineering. The preparation method comprises the following steps of: taking colibacillus CCTCC (colibacillus China center for type culture collection) NO: M208088 as a starting strain, adding a fermentation medium into a fermentation tank, fermenting for 2-6h, then adding a fed batch culture medium, and adding lactose in a flowing way at the mid-later period of the fermentation; and centrifuging the fermentation liquid to obtain liquid supernatant 1 and a thallus, diluting the centrifuged thallus by adding water and splitting in a heating way, recentrifuging to obtain liquid supernatant 2, merging the liquid supernatant 1 and the liquid supernatant 2, and absorbing, desorbing and vacuum drying by ion exchange resin to obtain the 3'-sialic acid lactose. A mass of the 3'-sialic acid lactose can be prepared by the CCTCC NO: M208088. The 3'-sialic acid lactose can change the current situation that the sialyloligosaccharide component is less in exogenous adding substances in conventional baby milk, and can provide the guarantee to add more efficient sialic acid trophic factors into the baby milk.
Description
technical field [0001] The invention relates to a preparation method of sialyllactose, in particular to a preparation method of 3'-sialyllactose, belonging to the technical field of bioengineering. Background technique [0002] With the rapid development of modern life science technology, glycobiology has gradually become a new hot spot in the field of life science. Among the many sugars, sialic acid sugar raw materials have become one of the most concerned sectors. Sialic acid is generally located at the end of the sugar chain in the body. This position is usually the sensitive end of physiological reactions and the first contact site for the transmission of life information, and plays a pivotal role. At present, research has found that sialic acid is closely related to many physiological and pathological occurrences in the body, and life science has derived a new research field "sialic acid" aimed at revealing the biological functions of sialic acid substances. Biology" ...
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