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Applied Biochemistry and Biotechnology

, Volume 189, Issue 4, pp 1245–1261 | Cite as

Target Discovery of Novel α-l-Rhamnosidases from Human Fecal Metagenome and Application for Biotransformation of Natural Flavonoid Glycosides

  • Bin-Chun LiEmail author
  • Tian Zhang
  • Yan-Qin Li
  • Guo-Bin Ding
Article

Abstract

As a green and powerful tool, biocatalysis has emerged as a perfect alternative to traditional chemistry. The bottleneck during process development is discovery of novel enzymes with desired properties and independent intellectual property. Herein, we have successfully bioprospected three novel bacterial α-l-rhamnosidases from human fecal metagenome using a combinatorial strategy by high-throughput de novo sequencing combined with in silico searching for catalytic key motifs. All three novel α-l-rhamnosidases shared low sequence identities with reported (< 35%) and putative ones (< 57%) from public database. All three novel α-l-rhamnosidases were over-expressed as soluble form in Escherichia coli with high-level production. Furthermore, all three novel α-l-rhamnosidases hydrolyzed the synthetic substrate p-nitrophenyl α-l-rhamnopyranoside and natural flavonoid glycosides rutin and naringin with some excellent properties, such as high activity in acidic pH, high activity at low or high temperature, and good tolerance for alcohols and DMSO. Our findings would provide a convenient route for target discovery of the promising biocatalysts from the metagenomes for biotransformation and biosynthesis.

Keywords

α-l-Rhamnosidase  Flavonoid glycoside  High-throughput sequencing  Catalytic key motif  Human fecal metagenome  In silico searching 

Notes

Funding information

This work was supported by the National Natural Science Foundation of China (No. 31400684) and the Natural Science Foundation of Shanxi (No. 2014021030-3).

Compliance with Ethical Standards

Our study had been approved by the Committee on the Ethics of Human and Animal Experiments of Shanxi University

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12010_2019_3063_MOESM1_ESM.docx (1.3 mb)
ESM 1 (DOCX 1281 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of BiotechnologyShanxi UniversityTaiyuanChina

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