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
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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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