Production method of tacrolimus
A seed and Streptomyces hygroscopicus technology, applied in the field of biomedicine, can solve problems such as burden, heavy patients, and low yield
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example 1
[0013] The screening of example 1 fermentation medium
[0014] Prepare shake flask fermentation medium A, B, C respectively, wherein the composition of the medium is as follows:
[0015] A: Soluble starch 2.0g / L; yeast powder 1.2g / L; glycerin 1.0g / L; corn steep liquor powder 2.5g / L; ammonium sulfate 2.2g / L, K 2 HPO 4 2.1g / L; MnCL 2 0.002g / L; ZnSO 4 0.001g / L; pH 7.4;
[0016] B: soluble starch 2.0g / L; peanut powder 1.2g / L; glycerin 1.0g / L; corn steep liquor powder 2.5g / L; ammonium sulfate 2.2g / L, K 2 HPO 4 2.1g / L; MnCL 2 0.002g / L; ZnSO 4 0.001g / L; pH 7.4;
[0017] C: soluble starch 2.0g / L; yeast powder 1.2g / L; glucose 1.0g / L; corn steep liquor powder 2.5g / L; ammonium sulfate 2.2g / L, K 2 HPO 4 2.1g / L; MnCL 2 0.002g / L; ZnSO 4 0.001g / L; pH 7.4;
[0018] Pour 20 mL of the prepared medium into a 500 mL Erlenmeyer flask, sterilize in an autoclave at 120°C for 15 minutes, insert Streptomyces hygroscopicus (ACCC No.40417) after cooling, and culture at 30°C for 6 days. , m...
example 2
[0021] The screening of example 2 seed culture medium
[0022] Prepare seed medium A, B, and C respectively, wherein the composition of the medium is as follows:
[0023] A: soluble starch 2.5g / L; yeast powder 1.5g / L; glycerin 2.5g / L; corn steep liquor powder 2.5g / L; peanut meal 2.5g / L;, K 2 HPO 4 2.1g / L; MnCL 2 0.0015g / L; ZnSO 4 0.002g / L; pH 7.4;
[0024] B: soluble starch 2.5g / L; peanut powder 1.2g / L; glycerin 1.0g / L; corn steep liquor powder 2.5g / L; peanut meal 2.5g / L; K 2 HPO 4 2.1g / L; MnCL 2 0.0015g / L; ZnSO 4 0.002g / L; pH 7.4;
[0025] C: Soluble starch 2.0g / L; Yeast powder 1.2g / L; Glycerin 2.5g / L; Corn steep liquor dry powder 2.5g / L; K 2 HPO 4 2.1g / L; MnCL 2 0.0015g / L; ZnSO 4 0.002g / L; pH 7.4;
[0026] Pour 20 mL of the prepared medium into a 500 mL Erlenmeyer flask, sterilize in an autoclave at 120°C for 15 minutes, inoculate Streptomyces hygroscopicus (ACCC No. 40417) after cooling, and incubate at 30°C for 3 days. After the culture was over, they were tr...
example 3
[0032] Strain: Streptomyces hygroscopicus (ACCC No.40417)
[0033] Fermentation method: 70L fermenter, batch fermentation
[0034] Fermentation process: 70L fermenter, liquid volume 50L, fermentation temperature controlled at 30°C, air flow rate adjusted to 0.5vvm, fermenter speed to 350rpm, pH, dissolved oxygen and glucose concentrations were recorded after 4 hours of fermentation, and The concentration of the fermentation product tacrolimus was measured after 120 hours of fermentation until the end of 144 hours of fermentation.
[0035] Fermentation results: after the fermentation, the measured concentration of tacrolimus was 250g / L, and the concentration of substrate glucose was 8g / L. The pH dropped from the initial 7.4 to 6.5, and the dissolved oxygen dropped from the initial 100% to 20%.
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