title: Highly efficient synthesis of endomorphin-2 under thermodynamic control catalyzed by organic solvent stable proteases with in situ product removal.
abstract: An efficient enzymatic synthesis of endomorphin-2 (EM-2) was achieved using organic solvent stable proteases in nonaqeous media, based on thermodynamic control and an in situ product removal methodology. The high stability of biocatalysts in organic solvents enabled the aleatoric modulation of the nonaqueous reaction media to shift thermodynamic equilibrium toward synthesis. Peptide Boc-Phe-Phe-NH2 was synthesized with a high yield of 96% by the solvent stable protease WQ9-2 in monophase medium with an economical molar ratio of the substrate of 1:1. The tetrapeptide Boc-Tyr-Pro-Phe-Phe-NH2 was synthesized with a yield of 88% by another organic solvent tolerant protease PT121 from Boc-Tyr-Pro-OH and Phe-Phe-NH2 in an organic-aqueous biphasic system. The reaction-separation coupling in both enzymatic processes provides "driving forces" for the synthetic reactions and gives a high yield and high productivity without purification of the intermediate, thereby making the synthesis more amenable to scale-up.
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, 30 Puzhunan Road, Nanjing, Jiangsu, China.
authors.forename: Jiaxing
authors.initials: J
authors.lastname: Xu
authors.forename: Honglin
authors.initials: H
authors.lastname: Sun
authors.forename: Xuejun
authors.initials: X
authors.lastname: He
authors.forename: Zhongzhong
authors.initials: Z
authors.lastname: Bai
authors.forename: Bingfang
authors.initials: B
authors.lastname: He
mesh_terms: D004795:Enzyme Stability; D006358:Hot Temperature; D009842:Oligopeptides / Q000138:chemical synthesis*; D009930:Organic Chemicals / Q000737:chemistry*; D010447:Peptide Hydrolases / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry*; D013816:Thermodynamics
chemical_list: D009842:Oligopeptides; D009930:Organic Chemicals; D010455:Peptides; D012997:Solvents; C105294:endomorphin 2; D010447:Peptide Hydrolases