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Bacterial strain and method for preparing glucosyl group apigenin by glycosylation in nonaqueous phase

A glucosyl apigenin, non-aqueous phase technology, applied in microorganism-based methods, biochemical equipment and methods, bacteria and other directions, can solve problems such as high cost, cumbersome steps, etc., and achieves low cost, simple separation, and glycosylation. product single effect

Inactive Publication Date: 2012-10-03
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the water solubility of glycosylated apigenin derivatives has been greatly improved compared with apigenin. At present, the preparation of glycosylated apigenin derivatives mainly adopts chemical synthesis, which is costly and complicated.

Method used

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  • Bacterial strain and method for preparing glucosyl group apigenin by glycosylation in nonaqueous phase
  • Bacterial strain and method for preparing glucosyl group apigenin by glycosylation in nonaqueous phase
  • Bacterial strain and method for preparing glucosyl group apigenin by glycosylation in nonaqueous phase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Screening and identification of organic solvent-resistant natural strains of glycosylated apigenin by biological method

[0036] Using sucrose as the carbon source for the growth of organic solvent-resistant bacteria, adding 10% to 25% (V / V) organic solvents such as methyl sulfoxide and dimethylformamide as the screening pressure, screening and separating from chemically polluted soil samples Obtaining resistance to organic solvents for extremophiles. The formulation of the screening medium was: sucrose 5 g / L, yeast extract 2.5 g / L, corn steep liquor 3 mL / L, KH 2 PO 4 2.0 g / L, MgSO 4 ·7H 2 O 1.0 g / L, initial pH 7.0.

[0037] Inoculate the screened microorganisms resistant to organic solvents into sieve medium, and culture them at 30° C. and 180 rpm for 12 to 24 hours to obtain fermentation broth or bacterial cell suspension. Configure reaction solution: dimethyl sulfoxide 5% (V / V), apigenin 0.05 g / L, sucrose or maltose or lactose 10 g / L, 50 mM pH 6.5 b...

Embodiment 2

[0040] Example 2 Fermentation of Staphylococcus saprophyticus CQ16 CCTCC M2012099 and Preparation of Resting Cells

[0041] Staphylococcus saprophyticus CQ16 CCTCC M2012099 was inoculated into the seed medium: yeast extract 5.0 g / L, peptone 10.0 g / L, NaCl 10.0 g / L, pH 7.0, cultured at 30°C, 180 rpm for 12 hours.

[0042] Expansion medium, fermentation medium, its components and contents are: sucrose 15g / L, yeast powder 10 g / L, KH 2 PO 4 0.8 g / L, CaCl 2 0.5g / L. The pH was adjusted to 8.0 with NaOH. The seed solution was inoculated into the expansion medium and fermentation medium at 0.5% (V / V), and cultivated at 30°C and 180 rpm for 12 hours.

[0043] After centrifuging at 8000-10000 rpm for 15 minutes, the bacterial cells were collected and washed 1-2 times with normal saline to obtain resting cells of Staphylococcus saprophyticus CQ16.

Embodiment 3

[0045] Resting cells of Staphylococcus saprophyticus CQ16 CCTCC M2012099 were obtained according to Example 2.

[0046] Configure reaction solution with dimethyl sulfoxide, apigenin, sucrose, phosphate buffer, the ratio of organic solvent dimethyl sulfoxide in the reaction solution is 15% (V / V), apigenin 0.2 g / L, the mole of phosphate buffer The concentration is 150 mmol / L, the pH of phosphate buffer is 8.3, and the ratio of sucrose to apigenin is 150:1 (w / w).

[0047] Disperse the bacteria obtained above in the reaction solution, add to the reactor, wet the cells at 1% (W / V), at 30 ℃, 180 rpm, react for 36 hours, then centrifuge at 8000-10000 rpm for 10 minutes to obtain the reaction solution The supernatant was analyzed by HPLC.

[0048] Take an appropriate amount of AB-8 macroporous adsorption resin, wet-pack the column (chromatographic column 40×600mm), pretreat the resin with 5% (V / V) hydrochloric acid, 2% sodium hydroxide solution, and wash with distilled water until ne...

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Abstract

The invention relates to a bacterial strain for preparing glucosyl group apigenin by glycosylation in a nonaqueous phase, which is Staphylococcus saprophyticus CQ16 with a collection number of CCTCC (China Center For Type Culture Collection) NO: M2012099. The invention also provides an application of the bacterial strain in preparing the glucosyl group apigenin and a method for preparing the glucosyl group apigenin by the glycosylation in the nonaqueous phase by the bacterial strain. According to the bacterial strain separated with the method disclosed by the invention, the apigenin can be efficiently catalyzed to prepare the glucosyl group apigenin. Meanwhile, the insoluble substrate apigenin can generate glycosylation reaction with a glucose donor in the presence of biological catalyst under relatively high concentration in the nonaqueous phase, reaction catalytic efficiency is improved, the problem that apigenin glucoside compound is scarce is solved, the solubility is improved, and a foundation is laid for further researching the biological activity of flavonoid compounds.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, relates to a strain and a method for preparing glucosyl apigenin, in particular to a strain and a method for preparing glucosyl apigenin by glycosylation in a non-aqueous phase. Background technique [0002] Flavonoids are a large class of phenolic compounds that widely exist in nature. These compounds are widely found in plants. So far, more than 5,000 kinds of plants have been found to contain flavonoids, and the types of flavonoids have reached more than 9,000. , the basic structure of flavones is 2-phenylchromone (2-phenylchromone), which has a C6-C3-C6 basic configuration. Specifically, it can be divided into flavonoids, flavonols, isoflavones, flavanones, and tea catechins. The general chemical structure is as follows: [0003] [0004] A large number of studies have shown that flavonoids have a wide range of biological activities and various pharmacological activities, suc...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/60C12R1/44
Inventor 何冰芳蒋微储建林吴斌柏中中
Owner NANJING UNIV OF TECH
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