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Method of synthesizing trehalose by virtue of whole cell catalysis

A trehalose, whole cell technology, applied in the field of functional sugar alcohol synthesis

Active Publication Date: 2013-06-12
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The technical problem to be solved by the present invention is to overcome the defects of existing cell permeability treatment methods, provide a new cell permeability treatment method, and improve the utilization of whole cells Catalyze the efficiency of trehalose synthesis, thereby further reducing the production cost of trehalose

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] This example illustrates the source of strains.

[0041] One of the strains used in this patent is derived from Streptomyces griseochromogenes Streptomyces griseochromogenes The Escherichia coli with the trehalose synthase gene, wherein the construction method of the strain adopts the method described in the patent CN201210160403, and the strain number is BL21-tre1.

[0042] Another bacterial species used in this patent is derived from Deinococcus desert Deinococcus deserti The Escherichia coli with the trehalose synthase gene, wherein the construction method of the strain adopts the method described in the patent CN2012101604029, and the strain number is BL21-tre2.

Embodiment 2

[0044] This example illustrates the specific synthesis steps and synthesis effects of catalytic synthesis of trehalose.

[0045] The BL21-tre1 strain obtained in Example 1 was used for trehalose synthesis catalysis.

[0046] The biosurfactant used in this example was purchased from sigma company, the product number is L511218-10MG, and it is a glycolipid biosurfactant with a purity of 90%-94%.

[0047] Firstly, the genetic engineering bacteria of Escherichia coli were cultivated on the solid LB medium with ampicillin by streaking method to obtain single colonies of Escherichia coli. The formula of the solid medium with ampicillin was: peptone 10 g / L, yeast powder 5 g / L, sodium chloride 10 g / L, ampicillin antibiotic 0.1 g / L, agar 20 g / L. The specific operation process is to prepare the medium according to the requirements, sterilize the prepared culture at 121 °C for 30 minutes, pour it on the plate after cooling, inoculate with the streaking method, and cultivate in a constan...

Embodiment 3

[0061] This example illustrates the specific synthesis steps and synthesis effects of the catalyzed synthesis of trehalose by cells without permeabilization treatment.

[0062] The BL21-tre1 strain obtained in Example 1 was used for trehalose synthesis catalysis.

[0063] In the implementation step, take the fermentation broth that has been induced to produce enzymes in Example 2 (complete the fourth step of Example 2), take the same operation as the fifth step of Example 2 medium-volume fermentation broth, and use an equal volume of The phosphate buffer replaced the buffer added with the ester peptide biosurfactant to treat the bacteria, and the subsequent implementation steps remained the same as in Example 2.

[0064] After calculation, the results showed that the conversion rate of maltose to cells treated by lipopeptide surfactant permeabilization was 8.2%.

[0065] From the comparison of Example 2 and Example 3, it can be found that the treatment of E. coli cells with b...

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Abstract

The invention relates to a method of synthesizing trehalose by virtue of whole cell catalysis. The method comprises the following steps that: recombinant Escherichia coli cells used for massively producing trehalose synthase are cultivated in a fermenting manner, permeable treatment is carried out on the recombinant cells by using an ecological biosurfactant, and the trehalose is synthesized by the treated cells in a phosphate buffering system by using maltose as a substrate. According to the method disclosed by the invention, the Escherichia coli after the permeable treatment is used for synthesizing the trehalose by using the 25-3 percent of maltose as the substrate, the conversion rate of the substrate can reach 55-60 percent after reaction for 16-20 hours at the temperature of 20-25 DEG C; in addition, compositions of a reaction solution are very simple, thus greatly simplifying an extraction and purification technology of the permeable treatment. By utilizing the method provided by the invention, the trehalose with high quality can be efficiently produced in a large scale with low cost.

Description

technical field [0001] The invention relates to a method for synthesizing trehalose by using whole cells to catalyze and belongs to the technical field of functional sugar alcohol synthesis. Background technique [0002] Trehalose (Trehalose), also known as Tan sugar, is a non-reducing disaccharide composed of glucose molecules connected by α,α-1,1-glycosidic bonds. It widely exists in microorganisms, shrimps, insects, vertebrates Among animals and plants. Trehalose has a good non-specific protective effect on organisms, biological macromolecules, and biofilms, making cells highly resistant to adverse environmental conditions (such as high temperature, freezing, dehydration, high osmotic pressure, etc.), so it is Known as "sugar of life". In addition, trehalose has the characteristics of stable properties, low sweetness, slow human absorption and no caries formation. Therefore, trehalose is widely used in the fields of food, medicine, cosmetics and other fine chemicals. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/12C12R1/19
Inventor 黄和江凌崔怀言唐飞李霜徐娴
Owner NANJING TECH UNIV
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