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- Title
Identification of a Novel Feruloyl Esterase by Functional Screening of a Soil Metagenomic Library.
- Authors
Li, Xuanxuan; Guo, Jia; Hu, Yimin; Yang, Yumeng; Jiang, Junwei; Nan, Fang; Wu, Shenglu; Xin, Zhihong
- Abstract
A cosmid metagenomic library containing 1.3 × 105 clones was created from a soil sample. A novel gene (fae-xuan) encoding a feruloyl esterase was identified through functional screening. Primary sequence analysis showed that the gene consisted of 759 base pairs and encoded a protein of 252 amino acids. The gene was expressed in Escherichia coli BL21 (DE3) and the corresponding purified recombinant enzyme exhibited a molecular weight of 29 kDa. The FAE-Xuan showed high activity (40.0 U/mg) toward methyl ferulate with an optimal temperature and pH of 30 °C and 5.0, respectively. Besides methyl ferulate, FAE-Xuan can also hydrolyze methyl sinapate and methyl p-coumarate. The substrate utilization preferences and phylogenetic analysis indicated that FAE-Xuan belongs to type A FAE. FAE-Xuan was quite stable over a broad pH range from 3.0 to 10.0. The activity reduced remarkably in presence of Cu2+. FAE-Xuan can enhance the quantity of ferulic acid from de-starched wheat bran in presence of xylanase. The work presented here highlighted the effectiveness of metagenomic strategy in identifying novel FAEs with diverse properties for potential use in industrial production.
- Subjects
METAGENOMICS; GENE expression; ESTERASES; FERULIC acid; WHEAT bran; XYLANASES; ESCHERICHIA coli
- Publication
Applied Biochemistry & Biotechnology, 2019, Vol 187, Issue 1, p424
- ISSN
0273-2289
- Publication type
Article
- DOI
10.1007/s12010-018-2832-1