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Antioxidant and Cytotoxic Effects and Identification of Ophiocordyceps sinensis Bioactive Proteins Using Shotgun Proteomic Analysis

Boon-Hong Kong1orcid tiny, Chee-Sum Alvin Yap1, Muhammad Fazril Mohamad Razif1orcid tiny, Szu-Ting Ng2, Chon-Seng Tan2 and Shin-Yee Fung1,3,4*orcid tiny

1Medicinal Mushroom Research Group, Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia

2LiGNO Biotech Sdn. Bhd., Jalan Perindustrian Balakong Jaya 2/2, Taman Perindustrian Balakong Jaya 2, 43300 Balakong Jaya, Selangor, Malaysia

3Centre for Natural Products Research and Drug Discovery (CENAR), University of Malaya, 50603 Kuala Lumpur, Malaysia

4University of Malaya Centre for Proteomics Research (UMCPR), University of Malaya, 50603 Kuala Lumpur, Malaysia

Article history:

Received: 21 January 2021

Accepted: 28 April 2021

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Key words:

Ophiocordyceps sinensis, antioxidant activity, cytotoxic effect, bioactive proteins, protein-polysaccharide complexes


Research background. Ophiocordyceps sinensis, a highly valued medicinal fungus, is close to extinction due to overexploitation. Successful cultivation of O. sinensis fruiting body (OCS02®) shows the cultivar has a promising nutritional value and numerous bioactive compounds. Antioxidant and antiproliferative properties and biologically active proteins of the OCS02® are investigated for possible development into nutraceuticals.

Experimental approach. The chemical composition of the OCS02® cold water extract was determined, and the antioxidant activities were examined using ferric reducing, DPPH· and O2·- scavenging assays. Tetrazolium dye (MTT) cytotoxic assay was performed to assess the antiproliferative activity of the extract. Bioactive proteins in the active fraction of the extract were identified using liquid chromatography (LC) and tandem-mass spectrometry (MS/MS). 

Results and conclusions. The OCS02® extract exhibited strong O2·- scavenging (expressed as Trolox equivalents (18.4±1.1) mol/g) and potent cytotoxic activities against adenocarcinomic human alveolar basal epithelial (A549) cells (IC50=(58.2±6.8) µg/mL). High molecular mass polysaccharides, proteins and protein-polysaccharide complexes could have contributed to the antioxidant and cytotoxic selectivity of the OCS02®. LC-MS/MS analysis identified several potential cytotoxic proteases and an oxalate decarboxylase protein which may exhibit protection effects on kidneys. 

Novelty and scientific contribution. The findings demonstrate the potential of OCS02® to be developed into functional food due to its promising superoxide anion radical scavenging capacity, cytotoxic effect and presence of bio-pharmaceutically active proteins.

*Corresponding author: +603 7967 5745
  +603 7967 4957

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