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

Journal Article · Sat Jan 01 04:00:00 EST 2005 · Acta Crystallographica. Section F

DOI:https://doi.org/10.1107/S1744309104031227· OSTI ID:22356098

Chandra, P. Manish ^[1]; Prabhune, Asmita; Pundle, Archana ^[1]; Turkenburg, Johan P.; Dodson, G. Guy ^[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)

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.

• OSTI ID: 22356098
• Journal Information: Acta Crystallographica. Section F, Journal Name: Acta Crystallographica. Section F Journal Issue: Pt 1 Vol. 61; ISSN ACSFCL; ISSN 1744-3091
• Country of Publication: United Kingdom
• Language: English

Related Subjects

75 CONDENSED MATTER PHYSICS
SUPERCONDUCTIVITY AND SUPERFLUIDITY
CRYSTALLIZATION
CRYSTALS
PRECURSOR
PROCESSING
PVA
SPACE GROUPS
SUBSTRATES