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Enzymatic rhamnosylation of anticancer drugs by an α-l-rhamnosidase from Alternaria sp. L1 for cancer-targeting and enzyme-activated prodrug therapy

  • Li Xu
  • Xiaohong Liu
  • Yinping Li
  • Zhenhao Yin
  • Lan Jin
  • Lili Lu
  • Jingyao Qu
  • Min XiaoEmail author
Biotechnological products and process engineering

Abstract

The synthesis of rhamnosylated compounds has gained great importance since these compounds have potential therapeutic applications. The enzymatic approaches for glycosylation of bioactive molecules have been well developed; however, the enzymatic rhamnosylation has been largely hindered by lacking of the glycosyl donor for rhamnosyltransferases. Here, we employed an α-l-rhamnosidase from Alternaria sp. L1 (RhaL1) to perform one-step rhamnosylation of anticancer drugs, including 2′-deoxy-5-fluorouridine (FUDR), cytosine arabinoside (Ara C), and hydroxyurea (Hydrea). The key synthesis conditions including substrate concentrations and reaction time were carefully optimized, and the maximum yields of each rhamnosylated drugs were 57.7 mmol for rhamnosylated Ara C, 68.6 mmol for rhamnosylated Hydrea, and 42.2 mmol for rhamnosylated FUDR. It is worth pointing out that these rhamnosylated drugs exhibit little cytotoxic effects on cancer cells, but could efficiently restore cytotoxic activity when incubated with exogenous α-l-rhamnosidase, suggesting their potential applications in the enzyme-activated prodrug system. To evaluate the cancer-targeting ability of rhamnose moiety, the rhamnose-conjugated fluorescence dye rhodamine B (Rha-RhB) was constructed. The fluorescence probe Rha-RhB displayed much higher cell affinity and cellular internalization rate of oral cancer cell KB and breast cancer cell MDA-MB-231 than that of the normal epithelial cells MCF 10A, suggesting that the rhamnose moiety could mediate the specific internalization of rhamnosylated compounds into cancer cells, which greatly facilitated their applications for cancer-targeting drug delivery.

Keywords

Rhamnosylation α-l-Rhamnosidase Alternaria sp. L1 Cancer-targeting Prodrug 

Notes

Funding information

This work was supported by the National Key Research and Development Program of China (2018YFA0902000), National Natural Science Foundation of China (31670062 and 31301155), the Science and Technology Development Project of Shandong Province (2016GGH4502 and 2017GSF21115), and the Fundamental Research Funds of Shandong University (2016JC028).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2019_10011_MOESM1_ESM.pdf (1.9 mb)
ESM 1 (PDF 1894 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.National Glycoengineering Research Center, Shandong Provincial Key Laboratory of Carbohydrate Chemistry and GlycobiologyShandong UniversityQingdaoPeople’s Republic of China
  2. 2.State Key Lab of Microbial TechnologyShandong UniversityQingdaoPeople’s Republic of China

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