Characterisation of novel angiotensin-I-converting enzyme inhibitory tripeptide, Gly-Val-Arg derived from mycelium of Pleurotus pulmonarius. (November 2017)

• Record Type: Journal Article
• Authors: Manoharan, Sivananthan; Shuib, Adawiyah Suriza; Abdullah, Noorlidah; Mohamad, Saharuddin Bin; Aminudin, Norhaniza
• Is Part Of: Process biochemistry. Volume 62(2017)
• Page Start: 215
• Page End: 222
• Publication Date: 2017-11
• Subjects: Angiotensin-I-converting enzyme -- Pleurotus pulmonarius -- ACE inhibitor -- Spontaneously hypertensive rats -- BIOPEP database; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2017.07.020
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: A novel ACE inhibitory tripeptide GVR was constructed through in silico analysis of Pleurotus pulmonarius protein sequence. In silico analysis indicated that the tripeptide GVR was not cleaved by trypsin, pepsin and alpha-chymotrypsin. Tripeptide GVR competitively inhibited ACE in vitro . Molecular docking analysis confirmed the ability of the tripeptide GVR to bind at the ACE's active site. Oral administration of the tripeptide GVR to SHRs was able to significantly reduce the systolic blood pressure of the rats. Abstract: It has been shown that fraction D6 from Pleurotus pulmonarius has the potential to inhibit ACE. After this discovery, additional studies were conducted to obtain peptides from that fraction, as ACE inhibitors. By size exclusion chromatography, single peak was resolved and termed as Psec. The IC50 of Psec was assessed to be 4.50 μg/mL, which was 2.5 times lower than that of D6 . When Psec was resolved by SDS-PAGE, three bands with estimated molecular weight of 63 kDa, 55 kDa and 11 kDa were observed. The protein bands were subjected to MALDI-Tof MS/MS for protein identification. By using the BIOPEP database for predicting in silico digestion of gastrointestinal (GI) enzymes, four stable tripeptides with ACE inhibitor potential resulting from GI enzyme digestion were identified, namely GVR, VVR, NPR, and VVL. The IC50 was estimated to be 55 μg/mL, 93 μg/mL, 110 μg/mL and >250 μg/mL individually. Based on a Lineweaver-Burk plot, tripeptide GVR was determined to be a competitive inhibitor and this was confirmed by molecular docking analysis. At 100 mg/kg of body weight (bw), the tripeptide GVR reduced SBP 33.5 mm Hg in SHRs. The results suggested that this tripeptide is potentially beneficial as an antihypertensive agent.