(Hyper)thermophilic Enzymes: Production and Purification

P. Falcicchio, M. Levisson, S.W.M. Kengen, S. Koutsopoulos

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)

Abstract

The discovery of thermophilic and hyperthermophilic microorganisms, thriving at environmental temperatures near or above 100 °C, has revolutionized our ideas about the upper temperature limit at which life can exist. The characterization of (hyper)thermostable proteins has broadened our understanding and presented new opportunities for solving one of the most challenging problems in biophysics: how is structural stability and biological function maintained at high temperatures where “normal” proteins undergo dramatic structural changes? In our laboratory we have purified and studied many thermostable and hyperthermostable proteins in an attempt to determine the molecular basis of heat stability. Here, we present methods to express such proteins and enzymes in E. coli and provide a general protocol for overproduction and purification. The ability to produce enzymes that retain their stability and activity at elevated temperatures creates exciting opportunities for a wide range of biocatalytic applications
LanguageEnglish
Pages487-496
JournalMethods in molecular biology
Volume1129
DOIs
Publication statusPublished - 2014

Fingerprint

Temperature
Enzymes
Proteins
Biophysics
Hot Temperature
Escherichia coli

Cite this

@article{019734423bfa4488831e2eac0808736f,
title = "(Hyper)thermophilic Enzymes: Production and Purification",
abstract = "The discovery of thermophilic and hyperthermophilic microorganisms, thriving at environmental temperatures near or above 100 °C, has revolutionized our ideas about the upper temperature limit at which life can exist. The characterization of (hyper)thermostable proteins has broadened our understanding and presented new opportunities for solving one of the most challenging problems in biophysics: how is structural stability and biological function maintained at high temperatures where “normal” proteins undergo dramatic structural changes? In our laboratory we have purified and studied many thermostable and hyperthermostable proteins in an attempt to determine the molecular basis of heat stability. Here, we present methods to express such proteins and enzymes in E. coli and provide a general protocol for overproduction and purification. The ability to produce enzymes that retain their stability and activity at elevated temperatures creates exciting opportunities for a wide range of biocatalytic applications",
author = "P. Falcicchio and M. Levisson and S.W.M. Kengen and S. Koutsopoulos",
year = "2014",
doi = "10.1007/978-1-62703-977-2_34",
language = "English",
volume = "1129",
pages = "487--496",
journal = "Methods in molecular biology",
issn = "1064-3745",
publisher = "Springer Verlag",

}

(Hyper)thermophilic Enzymes: Production and Purification. / Falcicchio, P.; Levisson, M.; Kengen, S.W.M.; Koutsopoulos, S.

In: Methods in molecular biology, Vol. 1129, 2014, p. 487-496.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - (Hyper)thermophilic Enzymes: Production and Purification

AU - Falcicchio, P.

AU - Levisson, M.

AU - Kengen, S.W.M.

AU - Koutsopoulos, S.

PY - 2014

Y1 - 2014

N2 - The discovery of thermophilic and hyperthermophilic microorganisms, thriving at environmental temperatures near or above 100 °C, has revolutionized our ideas about the upper temperature limit at which life can exist. The characterization of (hyper)thermostable proteins has broadened our understanding and presented new opportunities for solving one of the most challenging problems in biophysics: how is structural stability and biological function maintained at high temperatures where “normal” proteins undergo dramatic structural changes? In our laboratory we have purified and studied many thermostable and hyperthermostable proteins in an attempt to determine the molecular basis of heat stability. Here, we present methods to express such proteins and enzymes in E. coli and provide a general protocol for overproduction and purification. The ability to produce enzymes that retain their stability and activity at elevated temperatures creates exciting opportunities for a wide range of biocatalytic applications

AB - The discovery of thermophilic and hyperthermophilic microorganisms, thriving at environmental temperatures near or above 100 °C, has revolutionized our ideas about the upper temperature limit at which life can exist. The characterization of (hyper)thermostable proteins has broadened our understanding and presented new opportunities for solving one of the most challenging problems in biophysics: how is structural stability and biological function maintained at high temperatures where “normal” proteins undergo dramatic structural changes? In our laboratory we have purified and studied many thermostable and hyperthermostable proteins in an attempt to determine the molecular basis of heat stability. Here, we present methods to express such proteins and enzymes in E. coli and provide a general protocol for overproduction and purification. The ability to produce enzymes that retain their stability and activity at elevated temperatures creates exciting opportunities for a wide range of biocatalytic applications

U2 - 10.1007/978-1-62703-977-2_34

DO - 10.1007/978-1-62703-977-2_34

M3 - Article

VL - 1129

SP - 487

EP - 496

JO - Methods in molecular biology

T2 - Methods in molecular biology

JF - Methods in molecular biology

SN - 1064-3745

ER -