PDBsum entry 3iae

Lyase

PDB id: 3iae

Contents

Protein chains

554 a.a. *

Ligands

D7K ×2

Metals

_CA ×2

Waters ×210

* Residue conservation analysis

PDB id:

3iae

Name:

Lyase

Title:

Structure of benzaldehyde lyase a28s mutant with benzoylphosphonate

Structure:

Benzaldehyde lyase. Chain: a, b. Engineered: yes. Mutation: yes

Source:

Pseudomonas fluorescens. Organism_taxid: 294. Strain: biovar i. Gene: bznb. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.30Å R-factor: 0.207 R-free: 0.234

Authors:

G.S.Brandt,G.A.Petsko,D.Ringe,M.J.Mcleish

Key ref:

G.S.Brandt et al. (2010). Active-site engineering of benzaldehyde lyase shows that a point mutation can confer both new reactivity and susceptibility to mechanism-based inhibition. J Am Chem Soc, 132, 438-439. PubMed id: 20030408

Date:

13-Jul-09 Release date: 02-Mar-10

Protein chains

Q9F4L3  (Q9F4L3_PSEFL) -  Benzaldehyde lyase from Pseudomonas fluorescens

Seq: 563 a.a.

Struc: 554 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.4.1.2.38 - benzoin aldolase.
• Reaction: benzoin = 2 benzaldehyde
• Cofactor: Thiamine diphosphate

Thiamine diphosphate (Bound ligand (Het Group name = D7K) matches with 66.67% similarity)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

J Am Chem Soc 132:438-439 (2010)

PubMed id: 20030408

Active-site engineering of benzaldehyde lyase shows that a point mutation can confer both new reactivity and susceptibility to mechanism-based inhibition.

G.S.Brandt, M.M.Kneen, G.A.Petsko, D.Ringe, M.J.McLeish.

ABSTRACT

Benzaldehyde lyase (BAL) from Pseudomonas putida is a thiamin diphosphate (ThDP)-dependent enzyme that catalyzes the breakdown of (R)-benzoin. Here we report that a point mutant, BAL A28S, not only catalyzes the decarboxylation of benzoylformate but, like benzoylformate decarboxylase (BFDC), is also inactivated by the benzoylformate analogues methyl benzoylphosphonate (MBP) and benzoylphosphonate (BP). The latter has no effect on wild-type BAL, and the inactivation of the A28S variant is shown to result from phosphorylation of the newly introduced serine residue. This lends support to the proposal that an appropriately placed nucleophile facilitates the expulsion of carbon dioxide from the active site in many ThDP-dependent decarboxylases.