skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Cloning, preparation and preliminary crystallographic studies of penicillin V acylase autoproteolytic processing mutants

Abstract

The production, crystallization and characterization of three inactive mutants of penicillin V acylase from B. sphaericus in their respective precursor and processed forms are reported. The space groups are different for the native enzyme and the mutants. The crystallization of three catalytically inactive mutants of penicillin V acylase (PVA) from Bacillus sphaericus in precursor and processed forms is reported. The mutant proteins crystallize in different primitive monoclinic space groups that are distinct from the crystal forms for the native enzyme. Directed mutants and clone constructs were designed to study the post-translational autoproteolytic processing of PVA. The catalytically inactive mutants will provide three-dimensional structures of precursor PVA forms, plus open a route to the study of enzyme–substrate complexes for this industrially important enzyme.

Authors:
 [1]; ;  [1]; ;  [2]
  1. Division of Biochemical Sciences, National Chemical Laboratory, Pune 411008 (India)
  2. York Structural Biology Laboratory, Department of Chemistry, University of York, York YO10 5YW (United Kingdom)
Publication Date:
OSTI Identifier:
22356098
Resource Type:
Journal Article
Journal Name:
Acta Crystallographica. Section F
Additional Journal Information:
Journal Volume: 61; Journal Issue: Pt 1; Other Information: PMCID: PMC1952408; PMID: 16508111; PUBLISHER-ID: za5079; OAI: oai:pubmedcentral.nih.gov:1952408; Copyright (c) International Union of Crystallography 2005; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 1744-3091
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALS; PRECURSOR; PROCESSING; PVA; SPACE GROUPS; SUBSTRATES

Citation Formats

Chandra, P. Manish, Brannigan, James A., E-mail: jab@ysbl.york.ac.uk, Prabhune, Asmita, Pundle, Archana, Turkenburg, Johan P., Dodson, G. Guy, and Suresh, C. G., E-mail: jab@ysbl.york.ac.uk. Cloning, preparation and preliminary crystallographic studies of penicillin V acylase autoproteolytic processing mutants. United Kingdom: N. p., 2005. Web. doi:10.1107/S1744309104031227.
Chandra, P. Manish, Brannigan, James A., E-mail: jab@ysbl.york.ac.uk, Prabhune, Asmita, Pundle, Archana, Turkenburg, Johan P., Dodson, G. Guy, & Suresh, C. G., E-mail: jab@ysbl.york.ac.uk. Cloning, preparation and preliminary crystallographic studies of penicillin V acylase autoproteolytic processing mutants. United Kingdom. https://doi.org/10.1107/S1744309104031227
Chandra, P. Manish, Brannigan, James A., E-mail: jab@ysbl.york.ac.uk, Prabhune, Asmita, Pundle, Archana, Turkenburg, Johan P., Dodson, G. Guy, and Suresh, C. G., E-mail: jab@ysbl.york.ac.uk. 2005. "Cloning, preparation and preliminary crystallographic studies of penicillin V acylase autoproteolytic processing mutants". United Kingdom. https://doi.org/10.1107/S1744309104031227.
@article{osti_22356098,
title = {Cloning, preparation and preliminary crystallographic studies of penicillin V acylase autoproteolytic processing mutants},
author = {Chandra, P. Manish and Brannigan, James A., E-mail: jab@ysbl.york.ac.uk and Prabhune, Asmita and Pundle, Archana and Turkenburg, Johan P. and Dodson, G. Guy and Suresh, C. G., E-mail: jab@ysbl.york.ac.uk},
abstractNote = {The production, crystallization and characterization of three inactive mutants of penicillin V acylase from B. sphaericus in their respective precursor and processed forms are reported. The space groups are different for the native enzyme and the mutants. The crystallization of three catalytically inactive mutants of penicillin V acylase (PVA) from Bacillus sphaericus in precursor and processed forms is reported. The mutant proteins crystallize in different primitive monoclinic space groups that are distinct from the crystal forms for the native enzyme. Directed mutants and clone constructs were designed to study the post-translational autoproteolytic processing of PVA. The catalytically inactive mutants will provide three-dimensional structures of precursor PVA forms, plus open a route to the study of enzyme–substrate complexes for this industrially important enzyme.},
doi = {10.1107/S1744309104031227},
url = {https://www.osti.gov/biblio/22356098}, journal = {Acta Crystallographica. Section F},
issn = {1744-3091},
number = Pt 1,
volume = 61,
place = {United Kingdom},
year = {Sat Jan 01 00:00:00 EST 2005},
month = {Sat Jan 01 00:00:00 EST 2005}
}