通过培养基和发酵条件的优化提高红曲霉R”-30的洛伐他汀产量,并通过分离纯化制备高纯度的洛伐他汀。研究碳源、氮源、金属离子、培养基初始pH值、接种菌龄、接种量、装液量、摇床转速以及温度等对洛伐他汀产量的影响;进一步采用硅胶柱和Sephadex LH20柱层析对发酵液中洛伐他汀进行分离纯化。结果表明,最佳培养基配方为:甘油30 g/L,黄豆粉15 g/L,ZnSO4·7H2O 6.6 g/L,KH2PO4 0.17 g/L,MgSO4·7H2O 1.5 g/L;最适发酵条件为:培养基初始pH 5.0,接种龄64 h,接种量8%(体积分数),温度30 ℃,转速150 r/min,装液量50 mL/250 mL;在此条件下发酵12 d,洛伐他汀产量达615.3 mg/L,比优化前(312 mg/L)提高了97.2%;发酵液经NaOH碱提,柱层析分离纯化后,洛伐他汀纯度达93%,得率达45%。
This study aimed to improve the lovastatin yield with Monascus sp. R″-30 by optimizing the culture media and fermentation conditions and to prepare lovastatin with high purity by isolation and purification. The effects of carbon sources, nitrogen sources, metal ions, initial pH value of the culture medium, inoculation age, inoculation size, working fermentation volume, shaking speed, and temperature on lovastatin production were studied. The optimal medium formula for producing lovastatin were 30 g/L glycerol, 15 g/L soybean powder, 6.6 g/L ZnSO4·7H2O, 0.17 g/L KH2PO4 and 1.5 g/L MgSO4·7H2O. The optimal fermentation conditions were as follows: the initial pH 4.0, inoculation age of 64 hours, 8% inoculation size (V/V), fermentation temperature at 30 ℃, rotation speed at 150 r/min and the working volume of 50 mL/250mL (V/V). The maximal lovastatin production of 615.3 mg/L was obtained after 12 days’ fermentation, which was 97.2% increased. After separating and purifying the lovastatin in the beer by NaOH, silica gel column and LH20 column, the purity of lovastatin reached to 93% with the recovery of 45%.
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