Method for catalyzed synthesizing alpha arbutin from free cells or immobilized cells

A technology of immobilized cells and free cells, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problem of large equipment investment and operating costs, long cycle of α-arbutin, and α-arbutin Low yield and other problems, to achieve the effects of shortening the fermentation cycle, enhancing tolerance, and safe production operation

Inactive Publication Date: 2006-02-01
厦门绿启源生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. The follow-up treatment process of α-arbutin product produced by fermentation liquid is complicated, and the equipment investment and operation cost are relatively large;
[000

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] (a) Preparation of bacterial cells:

[0055] Pick 1 ring of Xanthomonas campestris CGMCC No.1243 strain preserved on the slant, add it to a liquid medium with a volume of 50ml, and cultivate it at 35°C and 130r / min on a shaker for 48h to obtain first-class seeds Then inoculate 10% of the primary seed liquid into the 500ml Erlenmeyer flask containing 200ml of the above-mentioned medium, and obtain the secondary seed liquid after cultivating on a 140r / min shaker at 35°C for 36h; use 10% of the secondary seed liquid The fermenter was inoculated, and the fermented liquid was centrifuged at 6000 r / min at 4° C. for 15 minutes to obtain Xanthomonas campestris CGMCC NO.1243 cells.

[0056] The seed liquid and fermenter medium are composed of the following components in parts by weight:

[0057] 60 parts of sucrose, (NH 4 ) 2 SO 4 2 parts, 0.5 parts of magnesium sulfate, 2 parts of sodium chloride, CaCl 2 2H 2 O 0.05 parts, ZnCl 2 0.01 parts, FeCl 2 4H 2 O 0.1 parts, N...

Embodiment 2

[0068] (a) Preparation of bacterial cells:

[0069] Repeat (a) step in Example 1 to prepare thalline cells;

[0070] (b) The reaction conditions for free cells to catalyze the synthesis of α-arbutin:

[0071] In 10mmol / L phosphate buffer solution, the free cell concentration is 200g / L, the reactant hydroquinone mass concentration is 60mmol / l, the sucrose mass concentration is 200mmol / l, the reaction temperature is 45°C, and the reaction time is 72 hours, the stirring speed was 200 rpm. Under the reaction conditions, the conversion rate of hydroquinone into α-arbutin reaches 92%, and 15 g of α-arbutin is contained in every liter of reaction solution.

[0072] (c) Recovery and repeated use of free cells:

[0073] After the catalyzed synthesis of α-arbutin by the free cells, the free cells can be recovered by centrifugation at 6000r / min, 4°C for 15 minutes, and the recovered cells are washed 2-3 times with 10mmol / L phosphate buffer solution, and put into the containing reactio...

Embodiment 3

[0080] (A) Preparation of bacterial cells:

[0081] Repeat (a) step in Example 1 to prepare thalline cells;

[0082] (B) Immobilization of bacterial cells:

[0083] Boil and dissolve sodium alginate with a concentration of 2% by mass in distilled water, mix it with an equal volume of bacterial suspension with a cell concentration of 150g / L, take a latex tube with an appropriate length of 4mm×6mm, and use an inner diameter of about 4.0mm dripper. Put the latex tube into the peristaltic pump, and adjust the speed of the peristaltic pump to 100 drops / min. Under the condition of stirring, add 0.1mol / L boric acid and 1.5mol / L CaCl dropwise to the mixed solution 2 In the mixed solution, immobilized gel beads with a uniform size are formed, and the maximum difference in diameter is ≤0.5 mm. After curing for 10 hours, the gel beads are taken out to obtain gel beads with a diameter of 3 mm.

[0084] (C) The reaction conditions for the catalytic synthesis of α-arbutin by immobilized c...

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Abstract

A process for catalytically synthesizing alpha-arbutoside from the free or immobilized cells of Xanthomonas campestris (CGMCC No.1243) includes such steps as preparing somatic cells, choosing reaction system, choosing immobilizing carrier, preparing immobilized cells, optimizing reaction condition, and extracting product. Said alpha-arbuto side can be used for beautifying skin.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a preparation method for catalytically synthesizing α-arbutin by using free cells or immobilized cells of Xanthomonas campestris CGMCC NO.1243. Background technique [0002] α-Arbutin can significantly inhibit the activity of tyrosinase, reduce the deposition of tyrosinase in the skin, and has the effects of bleaching, preventing discoloration and removing spots on the skin; it is an internationally recognized high-efficiency and safe It is an ideal whitening agent for whitening, freckle removing and sunscreen cosmetics. [0003] The technology of immobilized microbial cells (immobilized cells) is a new biotechnology developed in the 1960s, and its practical application effect is remarkable in chemical industry, fermentation production, energy, medicine and other industries. It is a method of using physical or chemical means to locate free microbial cells in a limited space area and k...

Claims

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Application Information

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IPC IPC(8): C12P33/00C12R1/01
Inventor 张鹏张淑荣刘春巧杨育红
Owner 厦门绿启源生物科技有限公司
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