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Bacillus subtilis for producing D-psicose as well as culture method and application thereof

A technology of Bacillus subtilis and psicose, which is applied in the field of Bacillus subtilis for producing D-psicose and its cultivation, can solve the problems of poor conversion ability, improve efficiency, reduce production cost, and adapt to the range of pH wide effect

Active Publication Date: 2022-03-22
BAOLINGBAO BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problem of poor conversion ability of D-psicose 3-epimerase in the preparation of D-psicose in the prior art, the present invention provides a Bacillus subtilis for producing D-psicose and Its cultivation method and application, the Bacillus subtilis bbl-12-B can high-yield D-psicose 3-epimerase, has the ability of high conversion fructose to generate D-psicose, improves production efficiency, and significantly reduce manufacturing cost

Method used

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  • Bacillus subtilis for producing D-psicose as well as culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Isolation and identification of strains

[0033] Plate culture medium: composed of the following components by weight percentage, 1% tryptone, 0.5% yeast extract powder, 1% sodium chloride, 2% agar powder, and the balance deionized water.

[0034] Slant culture medium: composed of the following components in weight percentage, 1% tryptone, 0.5% yeast extract powder, 1% sodium chloride, 2% agar powder, and the balance deionized water;

[0035] Shake flask culture medium: composed of the following components by weight percentage, peptone 2.5%, corn steep liquor powder 1%, glycerin 1%, anhydrous magnesium sulfate 0.05%, disodium hydrogen phosphate 0.05%, sodium dihydrogen phosphate 0.4%, the balance For deionized water, pH6.8;

[0036] (1) Strain source and screening

[0037] Select the soil near the special sugar workshop of Shandong Baolingbao Biological Co., Ltd., remove the topsoil with a shovel, take about 10g of the soil 10-15cm above the ground, and put it into a ...

Embodiment 2

[0045] Seed culture medium: composed of the following components by weight percentage, peptone 1%, yeast extract powder 0.5%, sodium chloride 1%, balance water, pH6.8;

[0046] Fermentation medium: composed of the following components by weight percentage, peptone 2.5%, corn steep liquor powder 1%, glycerin 1%, anhydrous magnesium sulfate 0.05%, disodium hydrogen phosphate 0.05%, sodium dihydrogen phosphate 0.4%, and the balance is Deionized water, pH6.8;

[0047] (1) Take Bacillus subtilis ( Bacillus subtilis ) blb-12-B, inoculated in the seed medium, and cultured for 10 hours at 37°C to obtain the seed solution;

[0048] (2) Take the seed solution prepared in step (1), inoculate it in the fermentation medium at a ratio of 5% by volume, ferment and cultivate it at 33° C., continuously replenish the fermentation medium during the fermentation process, and ferment for 52 hours to obtain Bacterial fermentation broth, centrifuged to obtain D-psicose 3-epimerase cells, then reco...

Embodiment 3

[0050] The effect of pH on the stability of enzyme activity:

[0051] For the D-psicose 3-epimerase serum obtained in Example 2, its enzyme activity at different pHs was tested to investigate the optimum pH of the enzyme. The reaction temperature was controlled at 55°C, respectively at pH4. The buffer solution of 0-8.5 configures the substrate to measure the enzyme activity, and the result is as follows figure 1 shown by figure 1 It can be seen that when the pH is between 5.5-7.5, the relative enzyme activity remains above 85%, when the pH value is between 5.0-8.0, the relative enzyme activity remains above 70%, and the enzyme activity is the highest at pH 6.5 , reaching 680U / ml.

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Abstract

The invention discloses bacillus subtilis for producing D-psicose as well as a culture method and application of the bacillus subtilis, the number of the bacillus subtilis is blb-12-B, the bacillus subtilis is preserved in the China General Microbiological Culture Collection Center (CGMCC) on December 8, 2021, and the preservation number is CGMCC No.24054. The invention further discloses a preparation method of the bacillus subtilis for producing D-psicose. The enzyme activity of D-psicose3-epimerase in the fermentation liquor of the bacillus subtilis obtained through separation is larger than or equal to 550 U / ml, the enzyme conversion efficiency is greatly improved, the production cost is remarkably reduced, when the pH value of the enzyme obtained through fermentation is 5.5-7.5, the enzyme activity is higher than 85%, the pH application range is wide, the color value and the conductivity of feed liquid can be reduced, the post-purification process of products is better facilitated, and the method is suitable for industrial production. The production efficiency of the product is improved, the production cost of the D-psicose is remarkably reduced, and industrial production control is facilitated.

Description

technical field [0001] The invention belongs to the technical field of microbes and their applications, and in particular relates to a bacillus subtilis for producing D-psicose and a culture method and application thereof. Background technique [0002] Rare sugar is a type of monosaccharide (the smallest unit of sugar) and sugar alcohol that exists in nature but is very small. Its taste is similar to sucrose, but it has low calories, high stability, sweetness and harmony, and no hygroscopicity. It can make up for the shortcomings of traditional sweeteners and play an important role in improving the diet of special groups. There are more than 50 rare sugars known so far. The typical representatives of rare sugar sweeteners are allulose and tagatose (respectively the C-3 and C-4 epimers of fructose), which not only have low calorie characteristics, but also inhibit blood sugar rise. High body fat accumulation, scavenging free radicals, neuroprotection, modifying drugs or acti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/02C12R1/125
CPCC12N1/20C12P19/02
Inventor 李克文薛雅莺熊小兰栾庆民张莉袁世英刘峰森茂治武昌
Owner BAOLINGBAO BIOLOGY
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