摘要
Objective:A novel protein was first purified from Panax quinquefolius L.(AGNP),and in vitro antioxidant activities of AGNP were first studied in this work.Methods:AGNP was purified by Ion-exchange chromatography and Gel-filtration chromatography.The chemical characterizations of AGNP were tested by sodium dodecyl sulfate–polyacrylamide gel electrophoresis(SDS-PAGE),high-pressure gel-filtration chromatography,MALDI-TOF-MS and HPLC.In vitro antioxidant effects were tested in simple antioxidant assay including 2,2-diphenylpicrylhydrazyl radical scavenging,superoxide radical(O2^-)scavenging,hydroxyl radical(OH)scavenging,and ferric-reducing ability.Results:The results showed which the content of AGNP measured by Bradford method was 2.42 mg/m L and the subunit molecular weight of AGNP measured by sodium dodecyl sulfate–polyacrylamide gel electrophoresis(SDS-PAGE)was 15 kD.The AGNP molecular weight was 15,114 Da both of SDS-PAGE and mass spectrum purity.The result of high-pressure gel-filtration chromatography demonstrated that the molecular weight of AGNP was 31,086 Da,which implied that AGNP was a homodimer.The in vitro Antioxidant results indicated that AGNP had obvious effects to remove the free radicals in vitro.Conclusion:In conclusion,AGNP had more powerful antioxidant capacity and it can be used as an effective natural antioxidant to alleviate oxidative stress.
Objective: A novel protein was first purified from Panax quinquefolius L.（AGNP）, and in vitro antioxidant activities of AGNP were first studied in this work. Methods: AGNP was purified by Ion-exchange chromatography and Gel-filtration chromatography. The chemical characterizations of AGNP were tested by sodium dodecyl sulfate–polyacrylamide gel electrophoresis（SDS-PAGE）, high-pressure gel-filtration chromatography, MALDI-TOF-MS and HPLC. In vitro antioxidant effects were tested in simple antioxidant assay including 2,2-diphenylpicrylhydrazyl radical scavenging, superoxide radical（O2-） scavenging, hydroxyl radical（OH） scavenging, and ferric-reducing ability. Results: The results showed which the content of AGNP measured by Bradford method was 2.42 mg/m L and the subunit molecular weight of AGNP measured by sodium dodecyl sulfate–polyacrylamide gel electrophoresis（SDS-PAGE） was 15 kD. The AGNP molecular weight was 15, 114 Da both of SDS-PAGE and mass spectrum purity. The result of high-pressure gel-filtration chromatography demonstrated that the molecular weight of AGNP was 31,086 Da, which implied that AGNP was a homodimer. The in vitro Antioxidant results indicated that AGNP had obvious effects to remove the free radicals in vitro. Conclusion: In conclusion, AGNP had more powerful antioxidant capacity and it can be used as an effective natural antioxidant to alleviate oxidative stress.
基金
financial support from the Jilin Provincial Key Laboratory of Biomacromolecules of Chinese Medicine, Local Standard Project of Chinese Medicinal Materials in Jilin Province (No. PZ-2016-03 and No. JLPZGF-2018-004)
The Science and Technology Development Plan Project of Jilin Province (No. 20190101010JH, No. 20190304095YY)
The National Key Research and Development Program of China (2017YFC1702106)
the National Traditional Chinese Medicine Standardization Project of China (No. ZYBZH-C-JL-22 and No. ZYBZH-C-HEB-11)
