在MTCC - 3538中进行化学诱变和高通量培养基优化可提高环孢菌素A的产量。
Chemical mutagenesis and high throughput media optimization in MTCC-3538 leads to enhanced production of cyclosporine A.
作者信息
Abrol Vidushi, Kushwaha Manoj, Mallubhotla Sharada, Jaglan Sundeep
机构信息
Fermentation & Microbial Biotechnology Division, Indian Institute of Integrative Medicine (CSIR), Canal Road, Jammu, 180001 India.
School of Biotechnology, Shri Mata Vaishno Devi University, Katra, Jammu, 182320 India.
出版信息
3 Biotech. 2022 Aug;12(8):158. doi: 10.1007/s13205-022-03219-x. Epub 2022 Jul 5.
UNLABELLED
Diethyl sulphate-based mutagenesis was performed on fungal strain MTCC-3538. Two mutant morphotypes MT1-3538 and MT2-3538 were selected for further chemo-profiling studies. LCMS/MS profiling of fungal crude extract confirmed that the wild-type and mutant strains (MT1-3538, MT2-3538) were competent to produce cyclosporine A. MT2-3538 produced 2.1 fold higher cyclosporine A in comparison to the wild type. Further, LCMS-based high throughput media optimization was performed for MT2-3538 in 20 different media combinations to increase cyclosporine A yield. On the basis of ion-intensity profiling, media combination consisting of Glucose 0.1 g/L; Peptone 0.005 g/L and Valine 0.005 g/L was selected and used for up-scaling purpose. Mutant MT2-3538 with optimized media combination increased cyclosporine yield 16 fold and could potentially be exploited for commercial outcomes.
SUPPLEMENTARY INFORMATION
The online version contains supplementary material available at 10.1007/s13205-022-03219-x.
未标记
对真菌菌株MTCC - 3538进行了基于硫酸二乙酯的诱变。选择了两种突变形态型MT1 - 3538和MT2 - 3538进行进一步的化学分析研究。真菌粗提物的液相色谱 - 质谱/质谱分析证实，野生型和突变菌株（MT1 - 3538、MT2 - 3538）都能产生环孢素A。与野生型相比，MT2 - 3538产生的环孢素A高2.1倍。此外，对MT2 - 3538在20种不同的培养基组合中进行了基于液相色谱 - 质谱的高通量培养基优化，以提高环孢素A的产量。根据离子强度分析，选择了由葡萄糖0.1 g/L、蛋白胨0.005 g/L和缬氨酸0.005 g/L组成的培养基组合用于扩大培养。具有优化培养基组合的突变体MT2 - 3538使环孢素产量提高了16倍，并且有可能用于商业生产。
补充信息
在线版本包含可在10.1007/s13205 - 022 - 03219 - x获取的补充材料。
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