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

IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY AMB Express Pub Date : 2024-02-15 DOI:10.1186/s13568-024-01679-3
Mengting Li, Shuhong Gao, Pengcheng Yang, Hejin Li
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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 KH2PO4) 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 KH2PO4, 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.

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提高 Cyberlindnera jadinii 的核糖核酸产量并优化发酵培养基。
为了提高核糖核酸(RNA)在工业应用中的生产率,本研究采用了菌株筛选和培养基优化的方法来提高 Cyberlindnera jadinii 的 RNA 含量。研究开发了一种结合常温和室温等离子体诱变、48深孔板发酵和微孔板阅读器检测的快速筛选方法。成功获得了高 RNA 含量的突变菌株 WB15,其 RNA 含量为 156 ± 4.5 mg/g DCW,是起始菌株 CCTCC AY 92020 的 1.4 倍。此外,通过普拉克特-伯曼设计和响应面方法,确定了影响 RNA 含量的三个重要因素(酵母提取物、大豆蛋白胨和 KH2PO4)。通过使用由 13.43 g/L 酵母提取物、12.12 g/L 大豆蛋白胨和 2.78 g/L KH2PO4 组成的最佳培养基,WB15 的 RNA 含量进一步提高到 184 ± 4.9 mg/g DCW。此外,与 AY 92020 相比,突变株 WB15 的细胞宽度更大,生长速度和单细胞 RNA 含量也分别增加了 22% 和 48.9%。核糖体组装的干扰,特别是核糖体蛋白与大亚基核糖体 RNA 比例的降低,可能间接导致 WB15 株系的 RNA 含量更高。总之,快速筛选与发酵培养基优化相结合被证明是提高 C. jadinii RNA 含量的有效方法,从而促进了 RNA 的工业化生产。
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来源期刊
AMB Express
AMB Express BIOTECHNOLOGY & APPLIED MICROBIOLOGY-
CiteScore
7.20
自引率
2.70%
发文量
141
审稿时长
13 weeks
期刊介绍: AMB Express is a high quality journal that brings together research in the area of Applied and Industrial Microbiology with a particular interest in ''White Biotechnology'' and ''Red Biotechnology''. The emphasis is on processes employing microorganisms, eukaryotic cell cultures or enzymes for the biosynthesis, transformation and degradation of compounds. This includes fine and bulk chemicals, polymeric compounds and enzymes or other proteins. Downstream processes are also considered. Integrated processes combining biochemical and chemical processes are also published.
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