Molecular cloning, expression and characterization of a novel feruloyl esterase from a soil metagenomic library with phthalate-degrading activity
To discover novel feruloyl esterases (FAEs) by the function-driven screening procedure from soil metagenome.
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.
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.
KeywordsFeruloyl esterase Function-driven screening Phthalate esters Site-directed mutagenesis Soil metagenome
This research was supported by the Fundamental Research Funds for the Central Universities (KYYJ201708) and Qing Lan Project of Jiangsu Province.
Compliance with ethical standards
Conflict of interest
The authors declare that there are no competing interests.
- Altschul SF, Wootton JC, Gertz EM et al (2005) Protein database searches using compositionally adjusted substitution matrices. FEBS J 272(20):5101–5109Google Scholar
- Amann RI, Ludwig W, Schleifer KH (1995) Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev 59:143–169Google Scholar
- Benjamin S, Pradeep S, Sarath Josh M et al (2015) A monograph on the remediation of hazardous phthalates. J Hazard Mater 298:58–72Google Scholar
- Brady SF (2007) Construction of soil environmental DNA cosmid libraries and screening for clones that produce biologically active small molecules. Nat Protoc 2:1297–1305Google Scholar
- Chen X, Zhang X, Yang Y et al (2015) Biodegradation of an endocrine-disrupting chemical di-n-butyl phthalate by newly isolated Camelimonas sp. and enzymatic properties of its hydrolase. Biodegradation 26:171–182Google Scholar
- Cheng F, Sheng J, Dong R et al (2012b) Novel xylanase from a holstein cattle rumen metagenomic library and its application in xylooligosaccharide and ferulic Acid production from wheat straw. J Agric Food Chem 60:12516–12524Google Scholar
- Haase-Aschoff P, Linke D, Nimtz M et al (2013) An enzyme from Auricularia auricula-judae combining both benzoyl and cinnamoyl esterase activity. Process Biochem 48:1872–1878Google Scholar
- Hancock JM, Bishop MJ (2004) Dictionary of bioinformatics and computational biology. ORF Finder (Open Reading Frame Finder). Wiley, New JerseyGoogle Scholar
- Handelsman J (2004) Metagenomics: application of genomics to uncultured microorganisms. Microbiol Mol Biol Rev 68:669–685Google Scholar
- Li X, Guo J, Hu Y et al (2018) Identification of a novel feruloyl esterase by functional screening of a soil metagenomic library. Appl Biochem Biotechnol 187(1):1–14Google Scholar
- Lu Y, Tang F, Wang Y et al (2009) Biodegradation of dimethyl phthalate, diethyl phthalate and di-n-butyl phthalate by Rhodococcus sp. L4 isolated from activated sludge. J Hazard Mater 168:938–943Google Scholar
- MacOn MB, Fenton SE (2013) Endocrine disruptors and the breast: early life effects and later life disease. J Mammary Gland Biol Neoplasia 18(1):43–61Google Scholar
- Ohlhoff CW, Kirby BM, Van Zyl L et al (2015) An unusual feruloyl esterase belonging to family VIII esterases and displaying a broad substrate range. J Mol Catal B Enzym 118:79–88Google Scholar
- Prates JA, Tarbouriech N, Charnock SJ et al (2001) The structure of the feruloyl esterase module of xylanase 10B from Clostridium thermocellum provides insights into substrate recognition. Structure 9(12):1183–1190Google Scholar
- Record E, Asther M, Sigoillot C et al (2003) Overproduction of the Aspergillus niger feruloyl esterase for pulp bleaching application. Appl Microbiol Biotechnol 62(4):349–355Google Scholar
- Reyes-Duarte D, Ferrer M, García-Arellano H (2012) Functional-based screening methods for lipases, esterases, and phospholipases in metagenomic libraries. Methods Mol Biol 861:101Google Scholar
- Schloss PD, Handelsman J (2005) Metagenomics for studying unculturable microorganisms: cutting the Gordian knot. Genome Biol 6:1–4Google Scholar
- Shu LS, Gang L, Xiao PH, Yu HL (2011) Molecular cloning, overexpression and characterization of a novel feruloyl esterase from a soil metagenomic library. J Mol Microbiol Biotechnol 20:196–203Google Scholar
- Tamura K, Stecher G, Peterson D et al (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30(12):2725–2729Google Scholar
- Torsvik V, Goksøyr J, Daae FL (1990) High diversity in DNA of soil bacteria. Appl Environ Microbiol 56:782–787Google Scholar
- Torsvik V, Daae FL, Sandaa RA, Ovreås L (1998) Novel techniques for analysing microbial diversity in natural and perturbed environments. J Biotechnol 64:53–62Google Scholar
- Wang L, Li Z, Zhu M et al (2016) An acidic feruloyl esterase from the mushroom Lactarius hatsudake: a potential animal feed supplement. Int J Biol Macromol 93:290–295Google Scholar
- Williamson G, Kroon PA, Faulds CB (1998) Hairy plant polysaccharides: a close shave with microbial esterases. Microbiology 144:2011–2023Google Scholar
- Wu M, Abokitse K, Grosse S et al (2012) New feruloyl esterases to access phenolic acids from grass biomass. Appl Biochem Biotechnol 168(1):129–143Google Scholar
- Yang J, Roy A, Zhang Y (2013a) BioLiP: a semi-manually curated database for biologically relevant ligand–protein interactions. Nucleic Acids Res 41:1096–1103Google Scholar
- Yang J, Roy A, Zhang Y (2013b) Protein–ligand binding site recognition using complementary binding-specific substructure comparison and sequence profile alignment. Bioinformatics 29:2588–2595Google Scholar
- Yu P, McKinnon JJ, Christensen DA (2011) Hydroxycinnamic acids and ferulic acid esterase in relation to biodegradation of complex plant cell walls. Can J Anim Sci. 85(3):255–267Google Scholar