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Biotechnol Lett. 2019 May 17. doi: 10.1007/s10529-019-02693-3. [Epub ahead of print]

Molecular cloning, expression and characterization of a novel feruloyl esterase from a soil metagenomic library with phthalate-degrading activity.

Author information

1
Key Laboratory of Food Processing and Quality Control, College of Food Science and Technology, Nanjing Agricultural University, Nanjing, 210095, People's Republic of China.
2
Key Laboratory of Food Processing and Quality Control, College of Food Science and Technology, Nanjing Agricultural University, Nanjing, 210095, People's Republic of China. xzhfood@njau.edu.cn.

Abstract

OBJECTIVES:

To discover novel feruloyl esterases (FAEs) by the function-driven screening procedure from soil metagenome.

RESULTS:

A novel FAE gene bds4 was isolated from a soil metagenomic library and over-expressed in Escherichia coli. The recombinant enzyme BDS4 was purified to homogeneity with a predicted molecular weight of 38.8 kDa. BDS4 exhibited strong activity (57.05 U/mg) toward methyl ferulate under the optimum pH and temperature of 8.0 and 37°C. Based on its amino acid sequence and model substrates specificity, BDS4 was classified as a type-C FAE. The quantity of the releasing ferulic acid can be enhanced significantly in the presence of xylanase compared with BDS4 alone from de-starched wheat bran. In addition, BDS4 can also hydrolyze several phthalates such as diethyl phthalate, dimethyl phthalate and dibutyl phthalate.

CONCLUSION:

The current investigation discovered a novel FAE with phthalate-degrading activity and highlighted the usefulness of metagenomic approaches as a powerful tool for discovery of novel FAEs.

KEYWORDS:

Feruloyl esterase; Function-driven screening; Phthalate esters; Site-directed mutagenesis; Soil metagenome

PMID:
31102076
DOI:
10.1007/s10529-019-02693-3

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