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Preparation method for high-purity D-psicose

一种阿洛酮糖、种子的技术，应用在高纯度D-阿洛酮糖的制备领域，能够解决酶活低、D-阿洛酮糖转化率低、易分解等问题，达到扩大应用范围、降低生产成本、生产成本降低的效果

Active Publication Date: 2017-03-22
SHANDONG BAILONG CHUANGYUAN BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the enzyme activity of the enzyme is still low and cannot meet the needs of actual production
[0005] However, at present, D-psicose has problems such as low conversion rate and easy decomposition, which greatly limits the application field of D-psicose, and it is difficult to effectively utilize the characteristics of the product itself

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] A Bacillus subtilis (Bacillus subtilis) BLCY-005 was preserved on October 26, 2016 in the General Microbiology Center of the China Committee for the Collection of Microbial Cultures, address: Microbiological Research, Chinese Academy of Sciences, No. 1, Yard 1, Beichen West Road, Chaoyang District, Beijing Institute, accession number CGMCC No.13152;

[0042] The original bacterial strain of Bacillus subtilis (Bacillus subtilis) BLCY-005 of the present invention was isolated from the soil near the production workshop of Bailong Chuangyuan, Dezhou, Shandong, and obtained after mutagenesis. The specific isolation process is as follows:

[0043] enrichment culture

[0044]Select the soil near the R&D pilot workshop of Bailong Chuangyuan, Dezhou, Shandong, remove the topsoil with a small shovel, take about 10g of soil 5-15cm above the ground, dilute it 10 times with sterile water, add LB medium for enrichment culture, Incubate at 30-38°C for 24 hours.

[0045] purebred seg...

Embodiment 2

[0057] The culture method of bacillus subtilis (Bacillus subtilis) BLCY-005 described in embodiment 1, the steps are as follows:

[0058] (a) Bacillus subtilis BLCY-005 was inoculated in LB medium, and activated for 12 hours at 35°C to obtain an activated strain;

[0059] (b) taking the activated bacterial strain prepared in step (a), inoculating it in the seed medium, and proliferating and culturing it for 12 hours at 35° C. to obtain the seed liquid;

[0060] Described seed culture medium component is as follows, is weight percent:

[0061] 1% peptone, 0.5% yeast extract powder, 1% sodium chloride, 0.01% anhydrous magnesium sulfate, 0.02% potassium dihydrogen phosphate, the rest water, pH6.8;

[0062] (c) taking the seed solution prepared in step (b), inoculating it in the fermentation medium at a ratio of 5% by volume, and expanding the cultivation for 48 hours at 35° C. to obtain the fermentation solution of Bacillus subtilis;

[0063] Described fermentation medium compo...

Embodiment 3

[0066] A preparation method of D-psicose, the steps are as follows:

[0067] (1) After centrifuging the fermentation liquid of Bacillus subtilis prepared in Example 2 for 30 minutes at 20° C. and a rotating speed of 3000 r / min, the thalline was homogenized for 10 minutes at a temperature of 20° C. and a pressure of 30 mpa to obtain A mixed solution containing D-psicose 3-epimerase;

[0068] (2) Prepare a fructose solution with a mass concentration of 60%, add the mixed solution containing D-psicose 3-epimerase, and add the mixed solution containing D-psicose 3-epimerase The addition amount is 5% of the volume percentage of the fructose solution, the pH value is adjusted to 5.5, and cobalt chloride is added in a proportion of 0.005% by mass, and the temperature is kept at 40°C for 30 hours, and then the fructose solution is added to the reaction solution to maintain the reaction The mass concentration of fructose in the solution is 20%, the reaction is continued for 30 hours, ...

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Abstract

The invention relates to a preparation method for high-purity D-psicose. The preparation method comprises the following steps: (1) subjecting the fermentation broth of Bacillus subtilis to centrifugation and then subjecting obtained thalli to homogenization so as to obtain a mixed solution containing D-psicose 3-epimerase; (2) preparing a fructose solution, adding the mixed solution containing D-psicose 3-epimerase, adjusting a pH value, carrying out a reaction under a heat insulation condition, then adding the fructose solution into the reaction solution in a fed-batch manner, continuing the reaction, and then stopping the reaction so as to prepare a crude D-psicose solution; and (3) subjecting the crude D-psicose solution to decoloring, filtering, ion exchange, chromatographic separation and concentration, and then to crystallization or drying so as to prepare D-psicose. The preparation method provided by the invention uses Bacillus subtilis fermentation broth for direct production and omits the process of enzyme extraction, so production cost is greatly reduced, a reduction of 25% or so compared with traditional production methods, so the competitiveness of prepared D-psicose is greatly improved.

Description

technical field [0001] The invention relates to a preparation method of high-purity D-psicose, which belongs to the field of biotechnology. Background technique [0002] D-Psicose is an important rare sugar, which exists in very small amounts in sugarcane molasses, dried fruits, sugar products, wheat and stingray plants in nature. Its name originates from the fact that a small amount of D-psicose can also be isolated from the antibiotic psicose adenosine. D-psicose is absorbed into the blood circulation through the small intestine in the human body, and will not be metabolized into energy after absorption in the small intestine, and has low fermentation availability for intestinal microorganisms. D-Psicose has a variety of important physiological functions: neuroprotection, reducing blood sugar, reducing fat, scavenging reactive oxygen species, anti-oxidation, inhibiting cancer cell proliferation, low-calorie sweetener, etc. [0003] D-Allulose can protect nerves by increa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/90C12P19/24C12P19/02
CPCC12N9/90C12P19/02C12P19/24C12Y503/01C07H1/08C07H3/02C12Y501/03C07B2200/13
Inventor 张明站干昭波窦光鹏邵先豹郝梅杜倩杨腾腾
Owner SHANDONG BAILONG CHUANGYUAN BIO TECH
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