5CT8

G158E/K44E/R57E/Y49E Bacillus subtilis lipase A with 0% [BMIM][Cl]


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 1.29 Å
  • R-Value Free: 0.143 
  • R-Value Work: 0.126 
  • R-Value Observed: 0.127 

wwPDB Validation   3D Report Full Report


This is version 1.2 of the entry. See complete history


Literature

Crystallographic Investigation of Imidazolium Ionic Liquid Effects on Enzyme Structure.

Nordwald, E.M.Plaks, J.G.Snell, J.R.Sousa, M.C.Kaar, J.L.

(2015) Chembiochem 16: 2456-2459

  • DOI: https://doi.org/10.1002/cbic.201500398
  • Primary Citation of Related Structures:  
    5CRI, 5CT4, 5CT5, 5CT6, 5CT8, 5CT9, 5CTA, 5CUR

  • PubMed Abstract: 

    We present the first crystallographic insight into the interactions of an ionic liquid (IL) with an enzyme, which has widespread implications for stabilizing enzymes in IL media for biocatalysis. Structures of Bacillus subtilis lipase A (lipA) and an IL-stable variant (QM-lipA) were obtained in the presence of increasing concentrations of 1-butyl-3-methylimidazolium chloride ([BMIM][Cl]). These studies revealed that the [BMIM] cation interacts with surface residues through hydrophobic and cation-π interactions. Of specific interest was the disruption of internal stacking interactions of aromatic side chains by [BMIM], which provides structural evidence for the mechanism of enzyme denaturation by ILs. The interaction of [BMIM] and Cl ions with lipA was reduced by the stabilizing mutations Y49E and G158E in QM-lipA. Ultimately, these findings present the molecular basis for stabilizing enzymes from IL-induced inactivation, as well as the selection of ILs that are less denaturing.


  • Organizational Affiliation

    Department of Chemical and Biological Engineering, University of Colorado, Campus Box 596, Boulder, CO, 80309, USA.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
Quadruple mutant lipase A180Bacillus subtilisMutation(s): 4 
Gene Names: lipAQX56_01625
UniProt
Find proteins for P37957 (Bacillus subtilis (strain 168))
Explore P37957 
Go to UniProtKB:  P37957
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupP37957
Sequence Annotations
Expand
  • Reference Sequence
Small Molecules
Ligands 1 Unique
IDChains Name / Formula / InChI Key2D Diagram3D Interactions
SO4
Query on SO4

Download Ideal Coordinates CCD File 
B [auth A]SULFATE ION
O4 S
QAOWNCQODCNURD-UHFFFAOYSA-L
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 1.29 Å
  • R-Value Free: 0.143 
  • R-Value Work: 0.126 
  • R-Value Observed: 0.127 
  • Space Group: P 21 21 21
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 48.09α = 90
b = 55.69β = 90
c = 64.56γ = 90
Software Package:
Software NamePurpose
PHENIXrefinement
iMOSFLMdata reduction

Structure Validation

View Full Validation Report



Entry History 

Deposition Data

Revision History  (Full details and data files)

  • Version 1.0: 2015-11-04
    Type: Initial release
  • Version 1.1: 2016-02-03
    Changes: Database references
  • Version 1.2: 2023-09-27
    Changes: Data collection, Database references, Derived calculations, Refinement description