Isolation and characterization of two novel gene encoded alkaline esterases from an alkaline soil metagenomic library

Yang Lu, Xiang Guo Liu, Ying Yu, He Zhi Qu, Shuo Yang, Bo Ning, Xiao Ping Wang, Dong Yun Hao

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

The development of industrial biotechnology has created an increasing demand for alkaline lipolytic enzymes with functional diversity. In this study, an alkaline soil metagenomic library was constructed to search for new lipolytic enzymes. Two novel gene encoded alkaline esterases(designated as estA and estB) were isolated by functional screening from the library. The estA gene consisted of 834 bp and coded for 277 amino acids with a molecular mass of 29998. Amino acid sequence homology analysis indicates that EstA belongs to α/β hydrolase fold family 4.4(abH4.4), with EstA being the smallest member of that family yet reported. The estB gene consisted of 1185 bp and encoded 394 amino acids with a theoretical molecular mass of 40090. Its conserved domain analysis shows that EstB belongs to the GDSL hydrolase superfamily. Both EstA and EstB exhibit only moderate identity(<38%) in amino acid sequence to the known lipolytic enzyme genes in the database. The two genes were respectively expressed in Escherichia coli and the protein products were purified for functional characterization. While the expressed EstA did not exhibit the functional properties that were superior to those of other esterases previously reported, the EstB was stable at temperature up to 45 °C and its maximum activity was measured to be 53.6 U/mg at pH=10. Both the enzymes have further enriched the diversity of the lipolytic enzymes database and also appear to be promising biocatalysts for potential biotechnological application.

Original languageEnglish (US)
Pages (from-to)583-590
Number of pages8
JournalChemical Research in Chinese Universities
Volume26
Issue number4
StatePublished - Dec 1 2010

Keywords

  • Alkaline esterase
  • Enzymatic characterization
  • Metagenome

ASJC Scopus subject areas

  • Chemistry(all)

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    Lu, Y., Liu, X. G., Yu, Y., Qu, H. Z., Yang, S., Ning, B., Wang, X. P., & Hao, D. Y. (2010). Isolation and characterization of two novel gene encoded alkaline esterases from an alkaline soil metagenomic library. Chemical Research in Chinese Universities, 26(4), 583-590.