Method for extracting L-alanine from L-alanine fermentation liquid
A technology of alanine and fermentation broth, which can be used in fermentation, chemical instruments and methods, preparation of organic compounds, etc., and can solve problems such as high cost, environmental pollution, and complicated steps.
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[0024] Preparation of fermentation broth: the preparation method of fermentation broth is as reported in the literature Xueli Zhang Kaemwich Jantama, Moore J C, et.al Production of L-alanine by metabolically engineered Eschierichia coli[J].Appl Microbiol Biotehnol, 2007, 77: 355- 366;
[0025] And Chinese patent 201110235159.8, a high-yield L-alanine XZ-A26 strain and its construction method and application. The specific method is as follows:
[0026] Strain: XZ-A26 strain preservation number (CGMCC No.4036)
[0027] The composition of seed medium and fermentation medium: glucose 120g / L, ammonium chloride 5g / L, NaH 2 PO 4 5g / L, Na 2 HPO 4 5g / L, MgSO 4 ·7H 2 O 1g / L, CaCl 2 22H 2 O 0.1g / L, trace inorganic salt 5ml / L, medium pH 6.5.
[0028] Composition of trace inorganic salts: FeCl 3 ·6H 2 O 1.5 mg, CoCl 2 ·6H 2 O 0.1 mg, CuCl 2 2H 2 O0.1mg, ZnCl 2 0.1 mg, Na 2 MoO 4 2H 2 O 0.1 mg, MnCl 2 4H 2 O 20.2mg, dilute to 1L with distilled water, filter to steriliz...
Embodiment 1
[0032] Fermented liquid 1.5L, L-alanine concentration 78.7mg / ml, after passing through 400000 molecular weight ultrafiltration membrane, obtain fermented liquid clear liquid, use H 2 SO 4 Adjust the pH of the supernatant to 5.0, add FeSO 4 ·7H 2 O 4.5g, after stirring and dissolving, keep stirring, add 30% H 2 o 210ml, and slowly raise the temperature to 50°C, maintain the temperature and stir for 3 hours, adjust the pH to 7.0 with NaOH solution, filter with diatomaceous earth, filter the reaction solution with suction and wash it with water to obtain a clear solution; add 5g of activated carbon to the clear solution, stir And raise the temperature to 80°C, keep it for 1h, remove the activated carbon by suction filtration to obtain the decolorized clear liquid, concentrate the clear liquid at 70°C under reduced pressure to 70ml, stir and crystallize the concentrated liquid for 8h, then suction filter to obtain the crystals, wash twice with deionized water , dried at 80° C....
Embodiment 2
[0034] Fermentation liquid 1.5L, L-alanine concentration 78.7mg / ml, add the mother liquor of crystallization of embodiment 1 and the combined liquid of crystal washing liquid, after crossing 100000 molecular weight ultrafiltration membrane, obtain fermented liquid supernatant liquid, use H 2 SO 4 Adjust the pH of the supernatant to 5.0, add FeSO 4 ·7H 2 O 4.5g, after stirring and dissolving, keep stirring, add 30% H 2 o 2 15ml, and slowly raised the temperature to 50°C, maintained the temperature and stirred for 3h, adjusted the pH to 7.0 with NaOH solution, filtered the reaction solution with diatomaceous earth, and washed the reaction solution with water to obtain a clear solution; add 5.3g of activated carbon to the clear solution, Stir and heat up to 80°C, maintain for 1h, remove the activated carbon by suction filtration to obtain a decolorized clear liquid, concentrate the clear liquid at 70°C under reduced pressure to 63ml, stir and crystallize the concentrated solut...
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