Cloning, purification, crystallization and preliminary structural studies of penicillin V acylase from Bacillus subtilis

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Priya RathinaswamyC G Suresh

Abstract

Penicillin acylase proteins are amidohydrolase enzymes that cleave penicillins at the amide bond connecting the side chain to their beta-lactam nucleus. An unannotated protein from Bacillus subtilis has been expressed in Escherichia coli, purified and confirmed to possess penicillin V acylase activity. The protein was crystallized using the hanging-drop vapour-diffusion method from a solution containing 4 M sodium formate in 100 mM Tris-HCl buffer pH 8.2. Diffraction data were collected under cryogenic conditions to a spacing of 2.5 A. The crystals belonged to the orthorhombic space group C222(1), with unit-cell parameters a = 111.0, b = 308.0, c = 56.0 A. The estimated Matthews coefficient was 3.23 A3 Da(-1), corresponding to 62% solvent content. The structure has been solved using molecular-replacement methods with B. sphaericus penicillin V acylase (PDB code 2pva) as the search model.

References

Apr 28, 1968·Journal of Molecular Biology·B W Matthews
Dec 31, 1995·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·K YoshidaY Fujita
May 13, 1999·Bioinformatics·P GouetF Métoz
May 20, 1999·Nature Structural Biology·C G SureshG G Dodson
Aug 31, 2004·Acta Crystallographica. Section D, Biological Crystallography·R Suresh KumarC G Suresh
Mar 2, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·P Manish ChandraC G Suresh

❮ Previous
Next ❯

Citations

Nov 29, 2014·Critical Reviews in Biotechnology·Vellore Sunder AvinashCheravakkattu Gopalan Suresh
Oct 13, 2009·Journal of Photochemistry and Photobiology. B, Biology·Atul KumarArchana Pundle
Feb 10, 2007·Journal of Biotechnology·Demin ZhangKazuhiro Nakanishi
May 2, 2015·International Journal of Biological Macromolecules·V S AvinashArchana Pundle
Jul 16, 2013·Biochemical and Biophysical Research Communications·V S AvinashSureshkumar Ramasamy
Oct 23, 2016·Biochimica Et Biophysica Acta. General Subjects·Deepak ChandSureshkumar Ramasamy
Dec 17, 2014·Applied and Environmental Microbiology·Jesús Torres-BaceteIsabel de la Mata
Apr 4, 2021·Microorganisms·Jack W DalyCormac G M Gahan

❮ 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.