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Homologous Expression and Characterization of α-L-rhamnosidase from Aspergillus niger for the Transformation of Flavonoids

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Abstract

Aspergillus niger has been used for homologous and heterologous expressions of many protein products. In this study, the α-L-rhamnosidase from A. niger (Rha-N1, GenBank XP_001389086.1) was homologously expressed in A. niger 3.350 by Agrobacterium tumefaciens-mediated transformation. The enzyme activity of Rha-N1 was 0.658 U/mL, which was obtained by cultivation of engineered A. niger in a 5-L bioreactor. Rha-N1 was purified by affinity chromatography and characterized. The optimum temperature and optimum pH for Rha-N1 were 60 °C and 4.5, respectively. Enzyme activity was promoted by Al3+, Li+, Mg2+, and Ba2+ and was inhibited by Mn2+, Fe3+, Ca2+, Cu2+, and organic solvents. The result indicated that rutin was the most suitable substrate for Rha-N1 by comparison with the other two flavonoid substrates hesperidin and naringin. The transformed products of isoquercitrin, hesperetin-7-O-glucoside, and prunin were identified by LC–MS and 1H-NMR.

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Funding

This research was financially supported by the National Natural Science Foundation of China (31600639), Science and Technology Project Founded by the Education Department of Jiangxi Province (GJJ202305), and Zhejiang University Student Science and Technology Innovation Activity Plan (**nMiao Talent Plan, 2021R403094).

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Authors and Affiliations

  1. Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People’s Republic of China

    Hangyu Ye, Luyuan Li, Yinjun Zhang & Jianyong Zheng

  2. Department of Fundamental Medicine, **nyu University, **nyu, 338004, People’s Republic of China

    **aojun Li

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  1. Hangyu Ye
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  2. **aojun Li
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  3. Luyuan Li
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Contributions

Jianyong Zheng and **aojun Li contributed to conceptualization; Hangyu Ye and **aojun Li performed data curation; Jianyong Zheng, **aojun Li, and Hangyu Ye contributed to funding acquisition; Hangyu Ye, Luyuan Li, and Yinjun Zhang performed investigation; Jianyong Zheng and **aojun Li were involved in supervision; Hangyu Ye and **aojun Li performed writing original draft.

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Correspondence to Jianyong Zheng.

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Ye, H., Li, X., Li, L. et al. Homologous Expression and Characterization of α-L-rhamnosidase from Aspergillus niger for the Transformation of Flavonoids. Appl Biochem Biotechnol 194, 3453–3467 (2022). https://doi.org/10.1007/s12010-022-03894-9

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