New thermophilic and thermostable esterase with sequence similarity to the hormone-sensitive lipase family, cloned from a metagenomic library

Jin Kyu Rhee, Dae Gyun Ahn, Yeon Gu Kim, Jong-Won Oh

Research output: Contribution to journalArticle

189 Citations (Scopus)

Abstract

A gene coding for a thermostable esterase was isolated by functional screening of Escherichia coli cells that had been transformed with fosmid environmental DNA libraries constructed with metagenomes from thermal environmental samples. The gene conferring esterase activity on E. coli grown on tributyrin agar was composed of 936 bp, corresponding to 311 amino acid residues with a molecular mass of 34 kDa. The enzyme showed significant amino acid similarity (64%) to the enzyme from a hyperthermophilic archaeon, Pyrobaculum calidifontis. An amino acid sequence comparison with other esterases and lipases revealed that the enzyme should be classified as a new member of the hormone-sensitive lipase family. The recombinant esterase that was overexpressed and purified from E. coli was active above 30°C up to 95°C and had a high thermal stability. It displayed a high degree of activity in a pH range of 5.5 to 7.5, with an optimal pH of approximately 6.0. The best substrate for the enzyme among the p-nitrophenyl esters (C 4 to C 16) examined was p-nitrophenyl caproate (C 6), and no lipolytic activity was observed with esters containing an acyl chain length of longer than 10 carbon atoms, indicating that the enzyme is an esterase and not a lipase.

Original languageEnglish
Pages (from-to)817-825
Number of pages9
JournalApplied and Environmental Microbiology
Volume71
Issue number2
DOIs
Publication statusPublished - 2005 Feb 1

Fingerprint

Sterol Esterase
Metagenomics
Esterases
esterases
hormone
hormones
enzyme
Enzymes
amino acid
Escherichia coli
enzymes
Lipase
ester
Pyrobaculum
Hot Temperature
esters
Metagenome
tributyrin
Amino Acids
hexanoic acid

All Science Journal Classification (ASJC) codes

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

Cite this

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title = "New thermophilic and thermostable esterase with sequence similarity to the hormone-sensitive lipase family, cloned from a metagenomic library",
abstract = "A gene coding for a thermostable esterase was isolated by functional screening of Escherichia coli cells that had been transformed with fosmid environmental DNA libraries constructed with metagenomes from thermal environmental samples. The gene conferring esterase activity on E. coli grown on tributyrin agar was composed of 936 bp, corresponding to 311 amino acid residues with a molecular mass of 34 kDa. The enzyme showed significant amino acid similarity (64{\%}) to the enzyme from a hyperthermophilic archaeon, Pyrobaculum calidifontis. An amino acid sequence comparison with other esterases and lipases revealed that the enzyme should be classified as a new member of the hormone-sensitive lipase family. The recombinant esterase that was overexpressed and purified from E. coli was active above 30°C up to 95°C and had a high thermal stability. It displayed a high degree of activity in a pH range of 5.5 to 7.5, with an optimal pH of approximately 6.0. The best substrate for the enzyme among the p-nitrophenyl esters (C 4 to C 16) examined was p-nitrophenyl caproate (C 6), and no lipolytic activity was observed with esters containing an acyl chain length of longer than 10 carbon atoms, indicating that the enzyme is an esterase and not a lipase.",
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New thermophilic and thermostable esterase with sequence similarity to the hormone-sensitive lipase family, cloned from a metagenomic library. / Rhee, Jin Kyu; Ahn, Dae Gyun; Kim, Yeon Gu; Oh, Jong-Won.

In: Applied and Environmental Microbiology, Vol. 71, No. 2, 01.02.2005, p. 817-825.

Research output: Contribution to journalArticle

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