Investigation, expression, and molecular modeling of ORF2, a metagenomic lipolytic enzyme

Applied Biochemistry and Biotechnology
Rosmeriana Afnis Marioto GarciaEliana Gertrudes de Macedo Lemos

Abstract

One clone exhibiting lipolytic activity was selected among 30 positives from a metagenomic library of a microbe consortium specialized in petroleum hydrocarbon degradation. From this clone, a sublibrary was constructed and a metagenome contig was assembled and analyzed using the ORF Finder; thus, it was possible to identify a potential ORF that encodes a lipolytic enzyme, denoted ORF2. This ORF is composed of 1035-bp 345 amino acids and displayed 98 % identity with an alpha/beta hydrolase from Pseudomonas nitroreducens (accession number WP024765380.1). When analyzed against a metagenome database, ORF2 also showed 76 % of sequence identity with a hypothetical protein from a marine metagenome (accession number ECT55726.1). The ProtParam analyses indicated that the recombinant protein ORF2 has a molecular mass approximately 39 kDa, as expected from its amino acid sequence, and based on phylogenetic analysis and molecular modeling, it was possible to suggest that ORF2 is a new member from family V. This enzyme exhibits the catalytic triad and conserved motifs typical from this family, wherein the serine residue is located in the central position of the conserved motif GASMGG. The orf2 gene was cloned in the expression vector pET28a...Continue Reading

References

Dec 5, 1993·Journal of Molecular Biology·A Sali, T L Blundell
Sep 1, 1997·Nucleic Acids Research·S F AltschulD J Lipman
Dec 23, 1999·Current Opinion in Structural Biology·M Nardini, B W Dijkstra
Aug 30, 2002·Applied and Environmental Microbiology·Doreen E GillespieJo Handelsman
Sep 27, 2002·Current Opinion in Biotechnology·Karl-Erich Jaeger, Thorsten Eggert
Jan 10, 2003·Nucleic Acids Research·David L WheelerLukas Wagner
Oct 11, 2003·Biotechnology Advances·R SharmaU C Banerjee
Jun 3, 2005·Nature Reviews. Microbiology·Patrick Lorenz, Jürgen Eck
May 5, 2006·Applied and Environmental Microbiology·C ElendW R Streit
Dec 16, 2006·Nucleic Acids Research·David L WheelerEugene Yaschenko
May 10, 2007·Molecular Biology and Evolution·Koichiro TamuraSudhir Kumar
Oct 22, 2008·Applied and Environmental Microbiology·Eun-Young KimJung-Hoon Yoon
Oct 30, 2008·Journal of Molecular Microbiology and Biotechnology·Helen L SteeleWolfgang R Streit
Mar 25, 2009·Journal of Microbiology and Biotechnology·Chao Wu, Baolin Sun
Dec 17, 2009·Trends in Biotechnology·Brajesh Kumar Singh
Jul 20, 2010·Bioresource Technology·Ren-Bao LiawChia-Yin Lee
Jul 19, 2011·Microbial Cell Factories·Arnaldo GlogauerNadia Krieger

❮ Previous
Next ❯

Citations

Feb 27, 2016·Applied Microbiology and Biotechnology·Thaís Carvalho MaesterEliana Gertrudes de Macedo Lemos
Jun 11, 2019·Frontiers in Pharmacology·Qirong LuMenghong Dai

❮ 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.