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提高嗜鞣管囊酵母中核糖核酸产量及优化发酵培养基

Improvement of ribonucleic acid production in Cyberlindnera jadinii and optimization of fermentation medium.

作者信息

Li Mengting, Gao Shuhong, Yang Pengcheng, Li Hejin

机构信息

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.

出版信息

AMB Express. 2024 Feb 15;14(1):24. doi: 10.1186/s13568-024-01679-3.

Abstract

To enhance the ribonucleic acid (RNA) productivity for industrial applications, this study employed strain screening and medium optimization to improve the content of RNA in Cyberlindnera jadinii. A rapid screening method, combining atmospheric and room temperature plasma mutagenesis, 48-deep-well plates fermentation, and microplate reader detection, was developed. A mutant strain named WB15 with high RNA content was successfully obtained, exhibiting the RNA content of 156 ± 4.5 mg/g DCW, 1.4 times of the starting strain CCTCC AY 92020. Furthermore, Plackett-Burman design and response surface methodology were employed to identify three significant factors (yeast extract, soybean peptone, and KHPO) affecting the RNA content. By utilizing the optimal medium composed of 13.43 g/L yeast extract, 12.12 g/L soybean peptone and 2.78 g/L KHPO, the RNA content of WB15 further increased to 184 ± 4.9 mg/g DCW. Additionally, the mutant strain WB15 exhibited a greater cellular width compared to AY 92020, along with increased growth rate and single-cell RNA content by 22% and 48.9%, respectively. Perturbations in ribosome assembly, specifically a reduction in the ratio of ribosomal proteins to ribosomal RNA of the large subunit, might indirectly contribute to the higher RNA content in the WB15 strain. Overall, the combination of rapid screening with fermentation medium optimization proved to be an effective approach for improving the RNA content of C. jadinii, thus facilitating the industrial production of RNA.

摘要

为提高工业应用中的核糖核酸(RNA)产量,本研究采用菌株筛选和培养基优化来提高嗜鞣管囊酵母中RNA的含量。开发了一种快速筛选方法,该方法结合了常压室温等离子体诱变、48孔板发酵和酶标仪检测。成功获得了一株名为WB15的RNA含量高的突变菌株,其RNA含量为156±4.5mg/g干细胞重,是出发菌株CCTCC AY 92020的1.4倍。此外,采用Plackett-Burman设计和响应面法来确定影响RNA含量的三个显著因素(酵母提取物、大豆蛋白胨和KHPO)。通过使用由13.43g/L酵母提取物、12.12g/L大豆蛋白胨和2.78g/L KHPO组成的优化培养基,WB15的RNA含量进一步提高到184±4.9mg/g干细胞重。此外,与AY 92020相比,突变菌株WB15的细胞宽度更大,生长速率和单细胞RNA含量分别提高了22%和48.9%。核糖体组装的扰动,特别是大亚基核糖体蛋白与核糖体RNA比例的降低,可能间接导致WB15菌株中较高的RNA含量。总体而言,快速筛选与发酵培养基优化相结合被证明是提高嗜鞣管囊酵母RNA含量的有效方法,从而促进了RNA的工业化生产。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cda7/10869677/c8e60dd40c82/13568_2024_1679_Fig1_HTML.jpg

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