A kind of method for separating and extracting epothilone b from fermented broth of Cystobacter celluli
A technology of S. cellulosus and epothilone, which is applied in the extraction of epothilone B and the field of separation and extraction of epothilone B, can solve the problems of low operation efficiency, low product yield, and high process cost. Achieve the effect of increasing production, high product purity and reducing production cost
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Embodiment 1
[0030] A method for isolating and extracting epothilone B from S. cellulosus fermentation broth, comprising the following steps:
[0031] 1. Fermentation and cultivation of S. cellulosus
[0032] Shake flask culture: first, inoculate the S. cellulosus strain ATCC25532 into a Erlenmeyer flask equipped with M26 liquid medium, and culture it on a shaker at 30°C and 200 rpm for 72 hours to obtain a shake flask seed solution;
[0033] Wherein, the composition of the M26 liquid medium is: potato starch 7.5-8.0g / L, soybean peptone 1.5-2.0g / L, glucose 1.5-2.0g / L, yeast powder 1.5-2.0g / L, MgSO 4 ·7H 2 O1.0~1.2g / L, CaCl 2 1.0~1.2g / L, EDTA-Fe 3+ 1mL / L, trace elements 1mL / L, pH 7.2;
[0034] Primary seed tank culture: Inoculate the obtained shake flask seed liquid into a 5L fermenter equipped with the above-mentioned M26 liquid medium, at a temperature of 30°C, a rotation speed of 150rpm, an air flow ratio of 1:0.5, and a tank pressure of 0.04~0.1kg / cm 2 , under the condition that t...
Embodiment 2
[0047] A method for isolating and extracting epothilone B from S. cellulosus fermentation broth, comprising the following steps:
[0048] 1. Fermentation and cultivation of S. cellulosus
[0049] Shake flask culture: first, inoculate the S. cellulosus strain ATCC25569 into an Erlenmeyer flask equipped with M26 liquid medium, and culture it on a shaker at 30°C and 180rpm for 84 hours to obtain a shake flask seed solution;
[0050] Wherein, the composition of described M26 liquid culture medium is identical with embodiment 1;
[0051] Primary seed tank cultivation: Inoculate the obtained shake flask seed liquid into a 5L fermenter equipped with the above-mentioned M26 liquid medium, at a temperature of 30°C, a rotation speed of 200rpm, an air flow ratio of 1:0.5, and a tank pressure of 0.05kg / cm 2 , under the condition that the pH value is 7.2, cultivate for 84 hours to obtain the first-grade strain liquid;
[0052] Secondary seed tank cultivation: inoculate vigorously growin...
Embodiment 3
[0064] A method for isolating and extracting epothilone B from S. cellulosus fermentation broth, comprising the following steps:
[0065] 1. Fermentation and cultivation of S. cellulosus
[0066] Shake flask culture: first, inoculate the S. cellulosus strain ATCC25532 into a Erlenmeyer flask equipped with M26 liquid medium, and culture it on a shaker at 30°C and 220rpm for 96 hours to obtain a shake flask seed solution;
[0067] Wherein, the composition of described M26 liquid culture medium is identical with embodiment 1;
[0068] Primary seed tank culture: Inoculate the obtained shake flask seed liquid into a 5L fermenter equipped with the above-mentioned M26 liquid medium, at a temperature of 30°C, a rotation speed of 160rpm, an air flow ratio of 1:0.5, and a tank pressure of 0.1kg / cm 2 , under the condition that the pH value is 7.2, cultivate for 96 hours to obtain the first-grade strain liquid;
[0069] Secondary seed tank cultivation: inoculate vigorously growing prim...
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