Effective separation of prolyl endopeptidase from Aspergillus Niger by aqueous two phase system and its characterization and application.

Effective separation of prolyl endopeptidase from Aspergillus Niger by aqueous two phase system and its characterization and application.

Int J Biol Macromol. 2020 Dec 18;:

Authors: Jiang B, Wang M, Wang X, Wu S, Li D, Liu C, Feng Z, Li J

Abstract
Aspergillus niger prolyl endopeptidase (An-PEP) has become a research focus because of its advantages in specifically cleaving the C-terminal peptide bond of proline residues, especially it was an industrial food-grade acidic PEP. Aqueous two-phase system (ATPS) was first applied for separating An-PEP from fermentation broth. Via response surface method (RSM) experiment, an effectively separation of An-PEP was achieved by ATPS containing27% (w/w) ethanol and 14.5% (w/w) (NH4)2SO4 at pH 6.0 with the recovery of 90.29 ± 0.23% and purification coefficient of 15.35 ± 0.30. The purified An-PEP was characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), fourier transform infrared (FTIR) and fluorescence spectrometry. The optimum temperature and pH of An-PEP were 40 °C and 4.5-5.0, respectively. An-PEP was activated and stabilized by Ca2+ but inhibited by Fe3+. The enzymatic application of purified An-PEP was evaluated by hydrolyzing egg white protein (EWP) to prepare bioactive peptides. The obtained hydrolysates had good scavenging ability of OH and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals, angiotensin converting enzyme (ACE) inhibitory activity and anti-gout activity. This research realized a low-cost, high-efficiency and simple separation technology of An-PEP and provided a broader idea for the preparation of bioactive peptides and the application of An-PEP.

PMID: 33347934 [PubMed – as supplied by publisher]

Source: Industry