Method for effectively producing 1,3-propanediol by modifying permeability of cell membrane
A propylene glycol and permeability technology, applied in 1 field, can solve the problems of long production cycle, low bacterial volume, low production intensity, etc., and achieve the effects of improving production efficiency, maintaining cell growth, and increasing permeability
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[0036] Bacteria: Klebsiella pneumoniae
[0037] Solid slant medium: beef extract 3g / L, MgSO 4 ·7H 2 O 0.2g / L, peptone 5g / L, NaCl 10g / L, agar 20g / L, glycerin 20g / L.
[0038] Seed medium: yeast extract 2g / L, MgSO 4 ·7H 2 O 0.2g / L, citric acid 0.42g / L, K 2 HPO 4 1.6g / L, K 2 HPO 4 3.4g / L, NH 4 Cl 3.0g / L, glycerol 30g / L, CaCl 2 0.5g / L.
[0039] Fermentation medium: glycerol 20-65g / L, K 2 HPO 4 ·3H 2 O 1.36g / L, NH 4 Cl 3.0g / L, yeast extract 2.0g / L, MgSO 4 ·7H 2 O 0.2g / L, CaCl 2 2.0g / L iron solution 1mL / L, inorganic ion solution 2mL / L, V B12 Solution 1mL / L, cobalt solution 1mL / L. Inorganic ions contain: CuCl 2 , H 3 BO 3 , MnCl 2 , Na 2 MoO 4 , NiCl 2 , ZnSO 4 , the content of trace elements is about 0.01-50mg / L.
[0040] Secondary seed preparation: take the fresh solid slant strain of Klebsiella pneumoniae, inoculate it in the seed medium, cultivate it on a shaker at 35°C at 150rpm for 24h, then inoculate it into the seed medium, and cultivate it at 35°...
Embodiment 1
[0044] 400ml of fermentation medium containing 45g / L industrial glycerin was placed in a 1L shake flask, and a volume of 40ml of secondary seeds was inserted, and the temperature was 37°C, and the shaker was cultivated at 180rpm. Add the concentration of 0%, 0.05%, 0.1%, 0.5%, 0.6%, 0.8%, 1% OP-10 respectively in the fermentation 8h, take samples at intervals, measure the cell concentration by spectrophotometer turbidimetry, the wavelength is 650nm , the cell concentration, that is, the amount of bacteria, is expressed as OD; the concentration of 1,3-propanediol is analyzed by gas chromatography. 24 hours to end fermentation.
[0045] Embodiment 1 has studied in fermenting glycerol to produce 1,3-propanediol technology, adds non-ionic surfactant OP-10, concentration is from 0~1.0% scope, different OP concentration is to Klebsiella pneumoniae growth and 1,3 - Effect of Propylene Glycol Yield. Figure 1 shows that at low concentrations, OP-10 mainly exhibits a promoting effect ...
Embodiment 2
[0048] 400ml of fermentation medium containing 50g / L industrial glycerol was placed in a 1L shake flask, and a volume of 40ml of secondary seeds was inserted, and the temperature was 37°C, and the shaker was cultivated at 180rpm. 0.1% OP-10 was added at 4h, 8h, 12h and 16h of fermentation, and the fermentation was finished in 24 hours. The concentration of 1,3-propanediol was analyzed by gas chromatography.
[0049]
[0050] Table 1 Fermentation 4h, 8h, 12h, 16h respectively added concentration of 0.1% OP-10 shake flask number
[0051] As can be seen from Figure 3, the addition of OP at different stages of fermentation has a greater impact on the output of 1,3-propanediol, and the effect of adding OP in 8-16 hours is the best, and the maximum output of 1,3-propanediol has increased by 12%. .
[0052] Through the above experiments, it can be proved that in the process of producing 1,3-propanediol by fermentation of glycerol, the method of increasing the permeability of the...
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