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The disruption of the MAPKK gene triggering the synthesis of flavonoids in endophytic fungus Phomopsis liquidambaris

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Abstract

Flavonoids, which are mainly extracted from plants, are important antioxidants and play an important role in human diseases. However, the growing market demand is limited by low productivity and complex production processes. Herein, the flavonoids biosynthesis pathway of the endophytic fungus Phomopsis liquidambaris was revealed. The mitogen-activated protein kinase kinase (MAPKK) of the strain was disrupted using a newly constructed CRISPR-Cas9 system mediated by two gRNAs which was conducive to cause plasmid loss. The disruption of the MAPKK gene triggered the biosynthesis of flavonoids against stress and resulted in the precipitation of flavonoids from fermentation broth. Naringenin, kaempferol and quercetin were detected in fed-batch fermentation with yields of 5.65 mg/L, 1.96 mg/L and 2.37 mg/L from P. liquidambaris for dry cell weigh using the mixture of glucose and xylose and corn steep powder as carbon source and nitrogen source for 72 h, respectively. The biosynthesis of flavonoids was triggered by disruption of MAPKK gene in P. liquidambaris and the mutant could utilize xylose.

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Acknowledgements

This study was supported by research grants from Key Technologies Research and Development Program of China (2018YFC1604206), National Natural Science Foundation of China (Grant Number: 32070058) and the National Key R&D Program of China (Grant Number: 2017YFD0800705). We thank Dr. Ling Lu (Nanjing Normal University) for kindly providing plasmid pX330 and FM-3.

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  1. Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, College of Life Sciences, Nanjing Normal University, Nanjing, 210023, Jiangsu, China

    Qian Yang, Mei Wu, Ya-Li Zhu, Ya-Qiong Yang, Yan-Zhen Mei & Chuan-Chao Dai

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  1. Qian Yang
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Correspondence to Yan-Zhen Mei or Chuan-Chao Dai.

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Yang, Q., Wu, M., Zhu, YL. et al. The disruption of the MAPKK gene triggering the synthesis of flavonoids in endophytic fungus Phomopsis liquidambaris. Biotechnol Lett 43, 119–132 (2021). https://doi.org/10.1007/s10529-020-03042-5

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