一株产L-苹果酸黑曲霉菌株的诱变筛选及发酵培养基优化
Mutagenesis Screening of a Producing L-Malic Acid Aspergillus niger Strain and Optimization of Fermentation Medium
刘书彤 1石冰冰 1谭奕阳 1厉成伟 2魏彩霞 2王德培 3薛鲜丽3
作者信息
- 1. 天津科技大学生物工程学院,天津 300457
- 2. 山东日照金禾博源生化有限公司,山东日照 276800
- 3. 天津科技大学生物工程学院,天津 300457;工业发酵微生物教育部重点实验室,天津 300457
- 折叠
摘要
为筛选一株产L-苹果酸能力强的黑曲霉菌株,通过紫外诱变与高浓度放线菌酮迭代诱变的方法,将野生型菌株诱变为一株产L-苹果酸能力强的黑曲霉菌株,并对其种子或发酵培养基成分进行优化.结果表明,诱变所得一株产L-苹果酸能力强的黑曲霉CGMCC NO.40550菌株,其摇瓶发酵时种子培养基最适葡萄糖浓度为 60 g/L、最适氮源为(NH4)2SO44.95 g/L,黑曲霉菌球形成最优转速220 r/min;发酵培养基最适葡萄糖和最适(NH4)2SO4浓度分别为180 g/L和4.95 g/L,CuSO4·5H2O为其生产苹果酸最佳微量元素,最适添加量为0.065 g/L.通过单因素实验的优化,优化后的培养基更有利于L-苹果酸的合成,发酵 96h时L-苹果酸产酸量达到 18.15 g/L,显著提高了245%,L-苹果酸占总酸的百分比达到71.69%.本研究为L-苹果酸的工业化生产奠定基础.
Abstract
In order to screen a strain of Aspergillus niger with high L-malic acid production ability,a wild-type strain was mutagenized into a strain with high L-malic acid production ability by iterative mutagenesis with UV mutagenesis and high concentration of actinomycinone,and the composition of its seed or fermentation medium was optimised.It showed that the optimal glucose concentration of the seed medium was 60 g/L,the optimal nitrogen source was(NH4)2SO4 with 4.95 g/L,and the optimal rotational speed for the formation of A.niger spheres was 220 r/min.The optimal concentration of glucose and(NH4)2SO4 of the fermentation medium were 180 g/L and 4.95 g/L,respectively.CuSO4·5H2O was the best trace element for the production of malic acid,with the optimal addition amount of 0.065 g/L.The optimized medium was more conducive to the synthesis of L-malic acid through the optimization of the one-way experiments.The results presented that the production of L-malic acid at 96 h reached 18.15 g/L,which was significantly improved by 245%,and the percentage of L-malic acid in the total acid reached 71.69%.This study lays the foundation for the industrial production of L-malic acid.
关键词
黑曲霉/紫外诱变/L-苹果酸/摇瓶发酵/培养条件Key words
Aspergillus niger/ultraviolet mutagenesis/L-malic acid/shake flask fermentation/culture conditions引用本文复制引用
基金项目
国家自然科学基金青年科学基金(31902193)
出版年
2024