Flavonoid-mediated inhibition of SARS coronavirus 3C-like protease expressed in Pichia pastoris

• Published in: Biotechnology letters Vol. 34; no. 5; pp. 831 - 838
• Main Authors: Nguyen, Thi Thanh Hanh, Woo, Hye-Jin, Kang, Hee-Kyoung, Nguyen, Van Dao, Kim, Young-Min, Kim, Do-Won, Ahn, Sul-Ah, Xia, Yongmei, Kim, Doman
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer-Verlag 01.05.2012; Springer Netherlands; Springer; Springer Nature B.V
• Subjects: Applied Microbiology; Biochemistry; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Catechin - analogs & derivatives; Catechin - metabolism; Cysteine Endopeptidases - biosynthesis; Cysteine Endopeptidases - chemistry; Cysteine Endopeptidases - genetics; drugs; energy; epigallocatechin; Flavonoids; Flavonoids - metabolism; Fundamental and applied biological sciences. Psychology; Gallates; Gene Expression; Hydrolysis; Inhibition; Inhibitory Concentration 50; Kinetics; Life Sciences; Microbiology; Molecular Weight; Original Research Paper; Pichia - genetics; Pichia pastoris; Protease; Protease Inhibitors - metabolism; Proteases; proteinases; quercetin; Recombinant; Recombinant Proteins - antagonists & inhibitors; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; SARS coronavirus; SARS Virus - enzymology; SARS Virus - genetics; Severe Acute Respiratory Syndrome; Viral Proteins - antagonists & inhibitors; Viral Proteins - biosynthesis; Viral Proteins - chemistry; Viral Proteins - genetics; Severe acute respiratory syndrome (SARS); Protease; Flavonoid; Molecular docking; Ascomycota; Enzyme; Severe acute respiratory syndrome; Pichia pastoris; Fungi; Infection; Virus; Peptidases; Coronavirus; Viral disease; Hydrolases; Inhibition; Coronaviridae; Nidovirales
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-011-0845-8

The 3C-like protease (3CLpro) of severe acute respiratory syndrome associated coronavirus (SARS-CoV) is vital for SARS-CoV replication and is a promising drug target. Recombinant 3CLpro was expressed in Pichia pastoris GS115 as a 42 kDa protein that displayed a K m of 15 ± 2 μM with Dabcyl-KTSAVLQSGFRKME-Edans as substrate. Purified 3CLpro was used for inhibition and kinetic assays with seven flavonoid compounds. The IC50 of six flavonoid compounds were 47–381 μM. Quercetin, epigallocatechin gallate and gallocatechin gallate (GCG) displayed good inhibition toward 3CLpro with IC50 values of 73, 73 and 47 μM, respectively. GCG showed a competitive inhibition pattern with K i value of 25 ± 1.7 μM. In molecular docking experiments, GCG displayed a binding energy of −14 kcal mol−1 to the active site of 3CLpro and the galloyl moiety at 3-OH position was required for 3CLpro inhibition activity.