Molecular cloning and characterization of a novel family VIII alkaline esterase from a compost metagenomic library

Biochemical and Biophysical Research Communications
Yong Ho KimHoon Kim

Abstract

A metagenomic library was constructed from completely fermented compost using a fosmid vector. From a total of 23,400 clones, 19 esterase-positive clones were selected on LB plates containing 1% glyceryl tributyrate as the substrate. The esterase gene of an esterase-positive clone, est2K, was on an ORF of 1299 bp and encoded a protein of 432 amino acids. Est2K had a SMTK motif and was a family VIII esterase. Unlike most family VIII esterases, Est2K had a signal peptide of 27 amino acids. The molecular mass and pI of the mature Est2K was calculated to be 44,668 Da and 4.48, respectively. The amino acid sequence of Est2K showed 72% identity with that of EstC, an esterase of an uncultured bacterium from leachate. The purified Est2K was optimally active at pH 10.0 and 50 degrees C. Est2K was stable in the presence of 30% methanol and exhibited a 2.4-fold higher activity in the presence of 5% methanol than in the presence of 1% isopropanol. Est2K preferred short to medium length p-nitrophenyl esters, especially p-nitrophenyl butyrate, as the substrate. Est2K did not hydrolyze beta-lactam antibiotics ampicillin and nitrocefin, even though Est2K showed the highest similarity to EstC.

References

Mar 11, 1992·Nucleic Acids Research·U J KimM I Simon
Feb 1, 1996·Applied and Environmental Microbiology·J ZhouJ M Tiedje
May 1, 1996·Applied and Environmental Microbiology·T BeffaM Aragno
Nov 1, 1996·Chemistry & Biology·J R KnoxJ M Frere
Oct 6, 1998·Applied and Environmental Microbiology·K NakasakiH Nagata
Sep 24, 1999·The Biochemical Journal·J L Arpigny, K E Jaeger
Apr 23, 2002·Journal of Applied Microbiology·S M Tiquia
Aug 22, 2002·Journal of the American Chemical Society·Sean F BradyJon Clardy
Sep 27, 2002·Current Opinion in Biotechnology·Karl-Erich Jaeger, Thorsten Eggert
Jun 30, 2004·Journal of Molecular Biology·Jannick Dyrløv BendtsenSøren Brunak
Sep 24, 2004·Journal of Molecular Microbiology and Biotechnology·Hiroyasu OginoHaruo Ishikawa
Dec 14, 2004·Microbiology and Molecular Biology Reviews : MMBR·Jo Handelsman
May 5, 2006·Applied and Environmental Microbiology·C ElendW R Streit
Sep 6, 2008·Applied Microbiology and Biotechnology·Jeong Ho JeonJung-Hyun Lee

❮ Previous
Next ❯

Citations

Feb 15, 2011·Applied Microbiology and Biotechnology·Eun Young YuJae Kwang Song
Nov 5, 2011·Extremophiles : Life Under Extreme Conditions·Xiao Ping HuDon Cowan
Sep 14, 2013·Extremophiles : Life Under Extreme Conditions·Ji-Eun ChoiJae Kwang Song
Feb 7, 2012·The Protein Journal·Charles C LeeWilliam J Orts
Jul 19, 2012·World Journal of Microbiology & Biotechnology·Diego Javier JiménezSandra Baena
Jan 5, 2012·Applied Biochemistry and Biotechnology·Yu Seok JeongHoon Kim
Nov 3, 2012·Applied Biochemistry and Biotechnology·Li-Ming OuyangJian-He Xu
Sep 3, 2013·Applied Biochemistry and Biotechnology·Changhyun Roh, Rolf D Schmid
Sep 13, 2011·Applied and Environmental Microbiology·Jeong Ho JeonJung-Hyun Lee
Jun 14, 2012·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Bum Han RyuT Doohun Kim
Feb 28, 2016·Enzyme and Microbial Technology·Mohammad Asadur RahmanZiduo Liu
Dec 17, 2014·Microbial Cell Factories·Viviane Paula MartiniNadia Krieger
Dec 5, 2015·Frontiers in Microbiology·Olalla López-LópezMaría-Isabel González-Siso
Aug 26, 2014·Enzyme and Microbial Technology·Shanshan ZhangZiduo Liu
Nov 5, 2016·International Journal of Biological Macromolecules·Yian HangZiduo Liu
Feb 3, 2012·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·V P MartiniN Krieger
May 14, 2020·Molecular Biology Reports·Francisco J AraujoVânia M M Melo
Feb 17, 2011·Journal of Agricultural and Food Chemistry·Hsu-Han ChuangJei-Fu Shaw

❮ 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.