Crystallization and preliminary X-ray crystallographic analysis of EstE1, a new and thermostable esterase cloned from a metagenomic library

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Jung-Sue ByunHyun-Soo Cho

Abstract

EstE1, a new thermostable esterase, was isolated by functional screening of a metagenomic DNA library from thermal environment samples. This enzyme showed activity towards short-chain acyl derivatives of length C4-C6 at a temperature of 303-363 K and displayed a high thermostability above 353 K. EstE1 has 64 and 57% amino-acid sequence similarity to est(pc)-encoded carboxylesterase from Pyrobaculum calidifontis and AFEST from Archaeoglobus fulgidus, respectively. The recombinant protein with a histidine tag at the C-terminus was overexpressed in Escherichia coli strain BL21(DE3) and then purified by affinity chromatography. The protein was crystallized at 290 K by the hanging-drop vapour-diffusion method. X-ray diffraction data were collected to 2.3 A resolution from an EstE1 crystal; the crystal belongs to space group P4(1)2(1)2, with unit-cell parameters a = b = 73.71, c = 234.23 A. Assuming the presence of four molecules in the asymmetric unit, the Matthews coefficient VM is calculated to be 2.2 A3 Da(-1) and the solvent content is 44.1%.

References

Apr 1, 1992·Protein Engineering·D L OllisJ Schrag
Sep 24, 1999·The Biochemical Journal·J L Arpigny, K E Jaeger
Dec 23, 1999·Current Opinion in Structural Biology·M Nardini, B W Dijkstra
Jul 31, 2002·Applied and Environmental Microbiology·Yuji HottaTadayuki Imanaka
Dec 17, 2002·Current Opinion in Biotechnology·Uwe T Bornscheuer
Apr 5, 2003·Bioresource Technology·G D Haki, S K Rakshit

❮ Previous
Next ❯

Citations

Jun 23, 2009·Extremophiles : Life Under Extreme Conditions·Mark LevissonServé W M Kengen
Dec 1, 2009·Biotechnology Journal·Marla TuffinDon A Cowan
May 21, 2013·Microbiological Research·Yanbing ZhuLuhong Hou

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Microbiology and Molecular Biology Reviews : MMBR
C Vieille, G J Zeikus
Acta Crystallographica. Section D, Biological Crystallography
T C Terwilliger
Proceedings of the National Academy of Sciences of the United States of America
Igor N Berezovsky, Eugene I Shakhnovich
© 2021 Meta ULC. All rights reserved