Method and bacterial strains for preparing (S)-epichlorohydrin through microbial transformation
A technology for the conversion of epichlorohydrin and microorganisms, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of severe reactions, low yields, and large pollution, and achieve low pollution and high yields , The effect of reducing raw material costs and production costs
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Embodiment 1
[0033] (1) Incline culture: medium: potato 200g / L, glucose 20g / L, agar 20g / L, adjust the pH to 7.0 with 1.0mol / L HCl or 1.0mol / L NaOH solution. After sterilizing at 121°C for 20min , cooling to prepare slant, inoculate Aspergillus niger CCTCC NO: M 2010275 strain stored in a 4°C refrigerator into the slant medium, and culture at 30°C until a single colony grows to obtain a slant;
[0034] (2) Seed culture: medium: glucose 20g / L, yeast extract powder 15g / L, peptone 15g / L, NaH 2 PO4· 2 h 2 O 1.0g / L, adjust the pH to 7.0 with 1.0mol / L HCl or 1.0mol / L NaOH solution. The filling volume is 50mL of liquid in a 250mL triangular flask. Sterilize at 121°C for 20 minutes, then cool the slanted seeds obtained in step (1) after sterilization, shake at 180r / min, and cultivate at 30°C for 24 hours as the seed solution;
[0035] (3) Fermentation culture: medium: starch 10g / L, bean cake powder 10g / L, KH 2 PO 4 1.0g / L, K 2 HPO 4 0.8g / L, MgSO 4 ·7H 2 O 0.4g / L, adjust the pH to 7.0 wi...
Embodiment 2
[0042] (1) Aspergillus niger (Aspergillus niger) CCTCC NO: M 2010275 According to the method in Example 1, after culturing and centrifuging to collect the thallus, mix the wet thallus and water at a mass ratio of 1:10 to prepare a bacterial suspension, and The bacterial suspension was mixed with 3% sodium alginate solution at a volume ratio of 1:2, and stirred evenly to obtain a sodium alginate mixture containing bacteria, and the mixed solution was dripped with 0.5% CaCl with a 0.5mm syringe needle 2 In the aqueous solution, solidify at 30°C for 10 hours, filter with sterile gauze, and rinse with 50mL sterile water to obtain immobilized cell particles;
[0043] (2) Biocatalysis: Measure about 1.5L of the prepared immobilized cell particles with a sterile graduated cylinder, and put them into a sterile fixed-bed bioreactor (S212-S-1L, Shanghai Hongjing Bio-Instrument Manufacturing Co., Ltd.). The bed of immobilized cell particles, the total volume of the reactor is 2.0L, and t...
Embodiment 3
[0048] (1) Aspergillus niger (Aspergillus niger) CCTCC NO: M 2010275 According to the method in Example 1, after culturing and centrifuging to collect the thalline, mix the wet thallus with water at a mass ratio of 1:15 to prepare a bacterial suspension, and The bacterial suspension was mixed with 5% sodium alginate solution at a volume ratio of 1:2, and stirred evenly to obtain a sodium alginate mixture containing bacteria, and the mixed solution was dripped with 1% CaCl with a 0.5mm syringe needle 2 In the aqueous solution, solidify at 30°C for 15 hours, filter with sterile gauze, and rinse with 50mL sterile water to obtain immobilized cell particles;
[0049] (2) Biocatalysis: Measure about 1.5L of the prepared immobilized cell particles with a sterile graduated cylinder, and put them into a sterile fixed-bed bioreactor (S212-S-1L, Shanghai Hongjing Bio-Instrument Manufacturing Co., Ltd.). The bed of immobilized cell particles, the total volume of the reactor is 2.0L, and t...
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