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采用常压室温等离子体诱变技术选育高产黄酮类化合物的鲍姆木层孔菌菌株及液体发酵验证

Employment of ARTP to Generate Phellinus baumii (Agaricomycetes) Strain with High Flavonoids Production and Validation by Liquid Fermentation.

作者信息

Zhang He-Nan, Jiang Fuchun, Qu Dehui, Wang Wen-Han, Dong Yating, Zhang Jing-Song, Wu Di, Yang Yan

机构信息

National Engineering Research Center of Edible Fungi, Key Laboratory of Applied Mycological Resources and Utilization of Ministry of Agriculture, Shanghai Key Laboratory of Agricultural Genetics and Breeding; Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Shanghai 201403, P.R. China.

Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, National Engineering Research Center of Edible Fungi, Key Laboratory of Edible Fungi Resources and Utilization (South), Ministry of Agriculture, No. 1000 Jinqi Road, Shanghai 201403, China.

出版信息

Int J Med Mushrooms. 2019;21(12):1207-1221. doi: 10.1615/IntJMedMushrooms.2019032976.

Abstract

To obtain Phellinus baumii strain with high flavonoids yield, ARTP was employed to generate mutants of a Ph. baumii strain, which were screened for higher flavonoids content. After five rounds of screening, four mutants were identified to produce more flavonoids than the wild type strain under optimal conditions, of which A67 was the mutant with the highest flavonoids productive capacity. When cultured in shake flasks, the maximum intracellular total flavonoids production of A67 reached 0.56 g/L, 86.67% higher than the total flavonoids in CK. Antagonistic testing, RAPD, and HPLC analysis suggested that ARTP caused changes of the genetic material and metabolites in Ph. baumii. In addition, the superiority of A67 to CK was proved by liquid fermentation using unstructured kinetic models, which was performed in a 50-L fermentor. The maximum intracellular total flavonoids production and dry mycelium weight of A67 reached 0.64 g/L and 15.24 g/L, which was an increase of 88.24% and 18.23% compared with CK, respectively. This work could provide an efficient and practical strategy to obtain high flavonoids production strains and the superiority of A67 could also provide a reference to further increase flavonoids production of Ph. baumii in large-scale production mode by submerged fermentation process.

摘要

为获得黄酮产量高的鲍姆木层孔菌菌株,采用常压室温等离子体诱变技术(ARTP)对一株鲍姆木层孔菌进行诱变,筛选黄酮含量更高的突变体。经过五轮筛选,鉴定出四个在最佳条件下比野生型菌株产生更多黄酮的突变体,其中A67是黄酮生产能力最高的突变体。在摇瓶培养时,A67的最大胞内总黄酮产量达到0.56 g/L,比对照(CK)中的总黄酮高出86.67%。拮抗试验、随机扩增多态性DNA(RAPD)分析和高效液相色谱(HPLC)分析表明,ARTP引起了鲍姆木层孔菌遗传物质和代谢产物的变化。此外,使用非结构动力学模型在50-L发酵罐中进行的液体发酵证明了A67相对于CK的优势。A67的最大胞内总黄酮产量和干菌丝体重量分别达到0.64 g/L和15.24 g/L,与CK相比分别增加了88.24%和18.23%。这项工作可为获得高黄酮产量菌株提供一种高效实用的策略,A67的优势也可为通过深层发酵工艺在大规模生产模式下进一步提高鲍姆木层孔菌的黄酮产量提供参考。

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