PDBsum entry 3qgo

Hydrolase

PDB id: 3qgo

Contents

Protein chain

316 a.a.

Ligands

0A9 ×3

DMS ×2

ACT

P6G ×2

Metals

_CA ×4

_ZN

Waters ×457

PDB id:

3qgo

Name:

Hydrolase

Title:

Structure of thermolysin in complex with l-phenylalanine methylester

Structure:

Thermolysin. Chain: a. Synonym: thermostable neutral proteinase. Ec: 3.4.24.27

Source:

Bacillus thermoproteolyticus. Organism_taxid: 1427

Resolution:

1.45Å R-factor: 0.112 R-free: 0.142

Authors:

G.Birrane,B.Bhyravbhatla,M.Navia

Key ref:

G.Birrane et al. (2014). Synthesis of Aspartame by Thermolysin: An X-ray Structural Study. Acs Med Chem Lett, 5, 706-710. PubMed id: 24944748 DOI: 10.1021/ml500101z

Date:

24-Jan-11 Release date: 04-Jan-12

Protein chain

P00800  (THER_BACTH) -  Thermolysin from Bacillus thermoproteolyticus

Seq: 548 a.a.

Struc: 316 a.a.

Enzyme reactions

• Enzyme class: E.C.3.4.24.27 - thermolysin.
• Reaction: Preferential cleavage: Xaa-|-Leu > Xaa-|-Phe.
• Cofactor: Ca(2+); Zn(2+)

DOI no: 10.1021/ml500101z

Acs Med Chem Lett 5:706-710 (2014)

PubMed id: 24944748

Synthesis of Aspartame by Thermolysin: An X-ray Structural Study.

G.Birrane, B.Bhyravbhatla, M.A.Navia.

ABSTRACT

Protease mediated peptide synthesis (PMPS) was first described in the 1930s but remains underexploited today. In most PMPS, the reaction equilibrium is shifted toward synthesis by the aqueous insolubility of product generated. Substrates and proteases are selected by trial and error, yields are modest, and reaction times are slow. Once implemented, however, PMPS reactions can be simple, environmentally benign, and readily scalable to a commercial level. We examined the PMPS of a precursor of the artificial sweetener aspartame, a multiton peptide synthesis catalyzed by the enzyme thermolysin. X-ray structures of thermolysin in complex with aspartame substrates separately, and after PMPS in a crystal, rationalize the reaction's substrate preferences and reveal an unexpected form of substrate inhibition that explains its sluggishness. Structure guided optimization of this and other PMPS reactions could expand the economic viability of commercial peptides beyond current high-potency, low-volume therapeutics, with substantial green chemistry advantages.