文章摘要
过程优化提高解脂亚洛酵母积累α-酮戊二酸
Process Optimization to Enhance the Accumulation of α-Ketoglutaric Acid in Yarrowia lipolytica
  
DOI:doi:10.3969/j.issn.1005-7021.2019.04.003
中文关键词: 解脂亚洛酵母  发酵优化  α-酮戊二酸  丙酮酸  溶氧  恒速补料
英文关键词: Yarrowia lipolytica  fermentation optimization  α-ketoglutaric acid  pyruvate  dissolved oxygen  constant feeding mode
基金项目:国家自然科学基金重大项目(21390204);国家自然科学基金项目(31770097)
作者单位
曾伟主 1.江南大学 工业生物技术教育部重点实验室江苏 无锡 2141222.江南大学 生物工程学院江苏 无锡 214122 
雷庆子 1.江南大学 工业生物技术教育部重点实验室江苏 无锡 2141222.江南大学 生物工程学院江苏 无锡 214122 
周景文 1.江南大学 工业生物技术教育部重点实验室江苏 无锡 2141222.江南大学 生物工程学院江苏 无锡 214122 
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中文摘要:
      目前,应用解脂亚洛酵母发酵生产α-酮戊二酸由于产量和底物转化率低、生产周期长等问题,仍未大规模工业化生产。为了解决这些问题,以研究室诱变选育获得的1株高产α-酮戊二酸的解脂亚洛酵母Yarrowia lipolytica WSH-Z06 C3为出发菌株,考察了该菌株在50 L发酵罐中转速、碳酸钙浓度、溶氧以及补料方式(多节点补料、恒速补料)等因素对α-酮戊二酸积累的影响。结果表明,当转速为300 r/min时,α-酮戊二酸和丙酮酸的产量分别为32.4 g/L和19.66 g/L;碳酸钙质量浓度为20 g/L时,α-酮戊二酸的产量提高至38.55 g/L,丙酮酸降低至8.28 g/L;控制溶氧水平在50%时,α-酮戊二酸产量为42.39 g/L,此时丙酮酸为6.22 g/L。比较高初始甘油浓度和不同的补料发酵策略,发现恒速补料效果最好,发酵144 h α-酮戊二酸产量达到66.27 g/L,丙酮酸产量为20.82 g/L。通过上述发酵过程参数的优化,α-酮戊二酸的产量和底物的转化率比未优化前分别提高了67.3%和4.56%,为解脂亚洛酵母工业化生产α-酮戊二酸提供一定参考。
英文摘要:
      Currently, problems of low output, low substrate conversion rate and long fermentation cycle restricted the industrial production of α-ketoglutaric acid (α-KG) by Yarrowia lipolytica in large scale. A mutant strain, Y. lipolytica WSH-Z06 C3, with capacity of high output of α-KG, obtained in previous research was taken as starting strain to solve these three problems. Accumulation effects of rotating speed, calcium carbonate concentration, dissolved oxygen and feeding methods (multi node feeding and constant speed feeding) and other factors on α-KG with Y. lipolytica WSH-Z06 C3 were investigated in 50 L fermentor. The results showed that when the rotating speed was set at300 r/min, the concentration of α-KG and pyruvate reached 32.4 g/L and 19.66 g/L respectively. The output of α-KG was increased to 38.55 g/L, and pyruvate decreased to 8.28 g/L; when the concentration of calcium carbonate was at 20 g/L. The output of α-KG was further increased to 42.39 g/L with pyruvate at 6.22 g/L by controlling the level of dissolved oxygen at 50%. With the strategies of high initial glycerol concentration and different feeding methods investigated, it was found that the constant feeding mode had the best effect. The production of α-KG and pyruvate reached 66.27 g/L and 20.82 g/L, respectively. The output of α-KG and pyruvate increased 67.3% and 456%, respectively, as compared with the process before optimization, and provided considerable reference for industrial production of α-KG with Y. lipolytica.
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