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 Citations (Scopus)

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
Externally publishedYes

Fingerprint

Esterases
Genes
Soils
Enzymes
Amino Acids
Molecular mass
Hydrolases
Escherichia coli Proteins
Biotechnology
Screening

Keywords

  • Alkaline esterase
  • Enzymatic characterization
  • Metagenome

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Isolation and characterization of two novel gene encoded alkaline esterases from an alkaline soil metagenomic library. / Lu, Yang; Liu, Xiang Guo; Yu, Ying; Qu, He Zhi; Yang, Shuo; Ning, Bo; Wang, Xiao Ping; Hao, Dong Yun.

In: Chemical Research in Chinese Universities, Vol. 26, No. 4, 01.12.2010, p. 583-590.

Research output: Contribution to journalArticle

Lu, Yang ; Liu, Xiang Guo ; Yu, Ying ; Qu, He Zhi ; Yang, Shuo ; Ning, Bo ; Wang, Xiao Ping ; Hao, Dong Yun. / Isolation and characterization of two novel gene encoded alkaline esterases from an alkaline soil metagenomic library. In: Chemical Research in Chinese Universities. 2010 ; Vol. 26, No. 4. pp. 583-590.
@article{56e045a40afa4df6b5368698c21ec8c9,
title = "Isolation and characterization of two novel gene encoded alkaline esterases from an alkaline soil metagenomic library",
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.",
keywords = "Alkaline esterase, Enzymatic characterization, Metagenome",
author = "Yang Lu and Liu, {Xiang Guo} and Ying Yu and Qu, {He Zhi} and Shuo Yang and Bo Ning and Wang, {Xiao Ping} and Hao, {Dong Yun}",
year = "2010",
month = "12",
day = "1",
language = "English (US)",
volume = "26",
pages = "583--590",
journal = "Chemical Research in Chinese Universities",
issn = "1005-9040",
publisher = "Jilin University Press",
number = "4",

}

TY - JOUR

T1 - Isolation and characterization of two novel gene encoded alkaline esterases from an alkaline soil metagenomic library

AU - Lu, Yang

AU - Liu, Xiang Guo

AU - Yu, Ying

AU - Qu, He Zhi

AU - Yang, Shuo

AU - Ning, Bo

AU - Wang, Xiao Ping

AU - Hao, Dong Yun

PY - 2010/12/1

Y1 - 2010/12/1

N2 - 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.

AB - 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.

KW - Alkaline esterase

KW - Enzymatic characterization

KW - Metagenome

UR - http://www.scopus.com/inward/record.url?scp=80555136043&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80555136043&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:80555136043

VL - 26

SP - 583

EP - 590

JO - Chemical Research in Chinese Universities

JF - Chemical Research in Chinese Universities

SN - 1005-9040

IS - 4

ER -