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利用鸟枪法蛋白质组学分析抗氧化和细胞毒性作用以及生物活性蛋白的鉴定

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

作者信息

Kong Boon-Hong, Yap Chee-Sum Alvin, Razif Muhammad Fazril Mohamad, Ng Szu-Ting, Tan Chon-Seng, Fung Shin-Yee

机构信息

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

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

出版信息

Food Technol Biotechnol. 2021 Jun;59(2):201-208. doi: 10.17113/ftb.59.02.21.7151.

Abstract

RESEARCH BACKGROUND

a highly valued medicinal fungus, is close to extinction due to overexploitation. Successful cultivation of fruiting body (OCS02®) shows that 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 O 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 O scavenging (expressed as Trolox equivalents (18.4±1.1) mol/g) and potent cytotoxic activities against adenocarcinomic human alveolar basal epithelial (A549) cells (IC=(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 CONTRIBUTIONS

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 biopharmaceutically active proteins.

摘要

研究背景

作为一种具有高价值的药用真菌,因过度开发而濒临灭绝。子实体(OCS02®)的成功栽培表明该品种具有可观的营养价值和众多生物活性化合物。对OCS02®的抗氧化和抗增殖特性以及生物活性蛋白进行了研究,以探讨其开发为营养保健品的可能性。

实验方法

测定了OCS02®冷水提取物的化学成分,并通过铁还原、DPPH和超氧阴离子清除试验检测了其抗氧化活性。采用四唑盐染料(MTT)细胞毒性试验评估提取物的抗增殖活性。使用液相色谱(LC)和串联质谱(MS/MS)鉴定提取物活性部分中的生物活性蛋白。

结果与结论

OCS02®提取物表现出较强的超氧阴离子清除能力(以Trolox当量表示为(18.4±1.1) μmol/g),并对人肺泡基底上皮腺癌(A549)细胞具有强大的细胞毒性活性(IC=(58.2±6.8) μg/mL)。高分子量多糖、蛋白质和蛋白质-多糖复合物可能促成了OCS02®的抗氧化和细胞毒性选择性。LC-MS/MS分析鉴定出几种潜在的细胞毒性蛋白酶和一种可能对肾脏具有保护作用的草酸脱羧酶蛋白。

新颖性与科学贡献

研究结果表明,OCS02®因其具有可观的超氧阴离子自由基清除能力、细胞毒性作用以及存在生物制药活性蛋白,具有开发成为功能性食品的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fef4/8284113/a3f192420957/FTB-59-201-f1.jpg

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