A thermostable esterase from Thermoanaerobacter tengcongensis opening up a new family of bacterial lipolytic enzymes

Lang Rao; Yanfen Xue; Cheng Zhou; Jin Tao; Gang Li; Jian R. Lu; Yanhe Ma

doi:10.1016/j.bbapap.2011.08.013

A thermostable esterase from Thermoanaerobacter tengcongensis opening up a new family of bacterial lipolytic enzymes

Lang Rao, Yanfen Xue, Cheng Zhou, Jin Tao, Gang Li, Jian R. Lu, Yanhe Ma

Research output: Contribution to journal › Article › peer-review

Abstract

An unidentified α/β hydrolase gene lipA3 from thermostable eubacterium species Thermoanaerobacter tengcongensis MB4 was cloned and heterologously expressed by Escherichia coli BL21(DE3)pLysS. The purified recombinant enzyme EstA3 turned out to be a monomeric thermostable esterase with optimal activity at 70 °C and pH 9.5. The enzyme showed lipolytic activity towards a wide range of ester substrates including p-nitrophenyl esters and triacylglycerides, with the highest activity being observed for p-nitrophenyl caproate at 150 U/mg and for Triacetin at 126 U/mg, respectively. Phylogenetic analysis revealed that EstA3 did not show homology to any identified bacterial lipolytic hydrolases. Sequence alignment showed that there was a common pentapeptide CHSMG with a cysteine replacing the first glycine in most esterase and lipase conserved motif GXSXG. The catalytic triad of EstA3 is Ser92, Asp269 and His292, which was confirmed by site directed mutagenesis. Based on the enzymatic properties and sequence alignment we concluded that the esterase EstA3 represented a novel bacterial lipolytic enzyme group and in chorological order this group was assigned as Family XIV. © 2011 Elsevier B.V. All Rights Reserved.

Original language	English
Pages (from-to)	1695-1702
Number of pages	7
Journal	Biochimica et Biophysica Acta - Proteins and Proteomics
Volume	1814
Issue number	12
DOIs	https://doi.org/10.1016/j.bbapap.2011.08.013
Publication status	Published - Dec 2011

Keywords

α/β hydrolase superfamily
Enzyme
Esterase
Extremophile
Lipolytic family
Thermophile

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10.1016/j.bbapap.2011.08.013

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title = "A thermostable esterase from Thermoanaerobacter tengcongensis opening up a new family of bacterial lipolytic enzymes",

abstract = "An unidentified α/β hydrolase gene lipA3 from thermostable eubacterium species Thermoanaerobacter tengcongensis MB4 was cloned and heterologously expressed by Escherichia coli BL21(DE3)pLysS. The purified recombinant enzyme EstA3 turned out to be a monomeric thermostable esterase with optimal activity at 70 °C and pH 9.5. The enzyme showed lipolytic activity towards a wide range of ester substrates including p-nitrophenyl esters and triacylglycerides, with the highest activity being observed for p-nitrophenyl caproate at 150 U/mg and for Triacetin at 126 U/mg, respectively. Phylogenetic analysis revealed that EstA3 did not show homology to any identified bacterial lipolytic hydrolases. Sequence alignment showed that there was a common pentapeptide CHSMG with a cysteine replacing the first glycine in most esterase and lipase conserved motif GXSXG. The catalytic triad of EstA3 is Ser92, Asp269 and His292, which was confirmed by site directed mutagenesis. Based on the enzymatic properties and sequence alignment we concluded that the esterase EstA3 represented a novel bacterial lipolytic enzyme group and in chorological order this group was assigned as Family XIV. {\textcopyright} 2011 Elsevier B.V. All Rights Reserved.",

keywords = "α/β hydrolase superfamily, Enzyme, Esterase, Extremophile, Lipolytic family, Thermophile",

author = "Lang Rao and Yanfen Xue and Cheng Zhou and Jin Tao and Gang Li and Lu, {Jian R.} and Yanhe Ma",

year = "2011",

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doi = "10.1016/j.bbapap.2011.08.013",

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T1 - A thermostable esterase from Thermoanaerobacter tengcongensis opening up a new family of bacterial lipolytic enzymes

AU - Rao, Lang

AU - Xue, Yanfen

AU - Zhou, Cheng

AU - Tao, Jin

AU - Li, Gang

AU - Lu, Jian R.

AU - Ma, Yanhe

PY - 2011/12

Y1 - 2011/12

N2 - An unidentified α/β hydrolase gene lipA3 from thermostable eubacterium species Thermoanaerobacter tengcongensis MB4 was cloned and heterologously expressed by Escherichia coli BL21(DE3)pLysS. The purified recombinant enzyme EstA3 turned out to be a monomeric thermostable esterase with optimal activity at 70 °C and pH 9.5. The enzyme showed lipolytic activity towards a wide range of ester substrates including p-nitrophenyl esters and triacylglycerides, with the highest activity being observed for p-nitrophenyl caproate at 150 U/mg and for Triacetin at 126 U/mg, respectively. Phylogenetic analysis revealed that EstA3 did not show homology to any identified bacterial lipolytic hydrolases. Sequence alignment showed that there was a common pentapeptide CHSMG with a cysteine replacing the first glycine in most esterase and lipase conserved motif GXSXG. The catalytic triad of EstA3 is Ser92, Asp269 and His292, which was confirmed by site directed mutagenesis. Based on the enzymatic properties and sequence alignment we concluded that the esterase EstA3 represented a novel bacterial lipolytic enzyme group and in chorological order this group was assigned as Family XIV. © 2011 Elsevier B.V. All Rights Reserved.

AB - An unidentified α/β hydrolase gene lipA3 from thermostable eubacterium species Thermoanaerobacter tengcongensis MB4 was cloned and heterologously expressed by Escherichia coli BL21(DE3)pLysS. The purified recombinant enzyme EstA3 turned out to be a monomeric thermostable esterase with optimal activity at 70 °C and pH 9.5. The enzyme showed lipolytic activity towards a wide range of ester substrates including p-nitrophenyl esters and triacylglycerides, with the highest activity being observed for p-nitrophenyl caproate at 150 U/mg and for Triacetin at 126 U/mg, respectively. Phylogenetic analysis revealed that EstA3 did not show homology to any identified bacterial lipolytic hydrolases. Sequence alignment showed that there was a common pentapeptide CHSMG with a cysteine replacing the first glycine in most esterase and lipase conserved motif GXSXG. The catalytic triad of EstA3 is Ser92, Asp269 and His292, which was confirmed by site directed mutagenesis. Based on the enzymatic properties and sequence alignment we concluded that the esterase EstA3 represented a novel bacterial lipolytic enzyme group and in chorological order this group was assigned as Family XIV. © 2011 Elsevier B.V. All Rights Reserved.

KW - α/β hydrolase superfamily

KW - Enzyme

KW - Esterase

KW - Extremophile

KW - Lipolytic family

KW - Thermophile

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DO - 10.1016/j.bbapap.2011.08.013

M3 - Article

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VL - 1814

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JO - Biochimica et Biophysica Acta - Proteins and Proteomics

JF - Biochimica et Biophysica Acta - Proteins and Proteomics

IS - 12

ER -

A thermostable esterase from Thermoanaerobacter tengcongensis opening up a new family of bacterial lipolytic enzymes

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Keywords

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