In the screening of antioxidant active substances for metabolites of entomogenous fungi, a strong antioxidant active substance was found in the solid culture of Paecilomyces gunnii strain RCEF4117. This study aimed to isolate and purify these active components and prepare pure products for structural identification. The antioxidant activity of methanol extracts from solid cultured of Paecilomyces gunnii mycelia was determined by 1,1-dipheny1-2-picryhydrazyl-thin layer chromatography (DPPH-TLC) and 1,1-dipheny1-2-picryhydrazyl hydrate-microplate (DPPH-Microplate). The active substances were preliminarily identified by high performance liquid chromatography-mass spectrometry DPPH (HPLC-MS-DPPH) and separated by reversed-phase high-performance liquid chromatography (RP-HPLC). We found that the methanol extract of the sample had strong antioxidant activity. Furthermore, two highly purified components, PG-1 (purity 97.31%, activity 89.62%) and PG-2 (purity 97.12%, activity 75.45%) were obtained from further separation and purification. The molecular formulas of PG-1 and PG-2 were C17H15NO6 and C16H14O6, respectively using HPLC-MS-DPPH analysis methods. In this study, two pure antioxidant components were successfully isolated and purified from Paecilomyces gunnii for the first time.
ZHANG Nengsheng
,
CHEN Xiaoling
,
HU Fenglin
. Purification and HPLC-MS analysis of antioxidant components in Paecilomyces gunnii[J]. Food and Fermentation Industries, 2020
, 46(16)
: 154
-158
.
DOI: 10.13995/j.cnki.11-1802/ts.024310
[1] 蒲蛰龙,李增智.昆虫真菌学[M].合肥:安徽科学技术出版社,1996.
[2] 黄红艳,刘九成.真菌杀虫剂在生物防治中的应用[J].北京农业,2015(6):73.
[3] 李增智.我国利用真菌防治害虫的历史、进展及现状[J].中国生物防治学报,2015,31(5):699-711.
[4] 董彩虹,李文佳,李增智,等.我国虫草产业发展现状、问题及展望——虫草产业发展金湖宣言[J].菌物学报,2016,35(1):1-15.
[5] 梁宗琦.中国真菌志(第三十二卷,虫草属)[M].北京:科学出版社,2007.
[6] LIANG Z Q, HAN Y F, CHU H L, et al. Studies on the genus Paecilomyces in China.I[J].Fungal Diversity,2005, 20: 83-101.
[7] 梁宗琦.古尼虫草分生孢子阶段的分离和鉴定[J].真菌学报,1985,4(3):32-36;67.
[8] 张泽文,傅岚,陈作红.古尼虫草无性型的分子鉴别[J].菌物学报,2005,24(3):344-348.
[9] 陈安徽,李春如,葛飞,等.古尼拟青霉小孢变种的主要有效成分分析[J].食品与发酵工业,2006,32(12):124-126.
[10] 甘卓亭,姚婷,丁璐.新古尼异虫草的化学成分、药理作用及开发应用研究进展[J].天然产物研究与开发,2019,31(6):1 109-1 115;1 108.
[11] 张倩. 古尼拟青霉主要成分的分析及药理作用初步研究[D].合肥:安徽农业大学,2011.
[12] ZHENG Y B, XU X P, ZOU X W. Biotransformation of caffeine in oolong tea by Paecilomyces gunnii[J]. International Biodeterioration & Biodegradation, 2016, 114: 141-144.
[13] LU R L, LIU X X, GAO S, et al. New tyrosinase inhibitors from Paecilomyces gunnii[J]. Journal of Agricultural and Food Chemistry, 2014, 62(49): 11 917-11 923.
[14] 章能胜. 一株古尼拟青霉提取物清除自由基活性研究[D].合肥:安徽农业大学,2010.
[15] ZHANG C, BERTIN R, FROLDI G. Ec 50 estimation of antioxidant activity in DPPH assay using several statistical programs[J]. Food Chemistry, 2013, 138(1): 414-420.
[16] DAWIDOWICZ A L, WIANOWSKA D, OLSZOWY M. On practical problems in estimation of antioxidant activity of compounds by DPPH method (Problems in estimation of antioxidant activity)[J]. Food Chemistry, 2012, 131(3): 1 037-1 043.
[17] CIES′LA L, KRYSZEN′ J, STOCHMAL A,et al.Approach to develop a standardized TLC-DPPH test for assessing free radical scavenging properties of selected phenolic compounds[J].Journal of Pharmaceutical and Biomedical Analysis, 2012, 70: 126-135.
[18] 胡丰林,陆瑞利,黄勃,等.一种被毛孢(Hirsutella sp.)培养液中自由基清除剂的分析、分离和制备[J].微生物学报,2008,48(8):1 035-1 041.
[19] LU R L, BAO G H, HU F L, et al. Comparison of cytotoxic extracts from fruiting bodies, infected insects and cultured mycelia of Cordyceps formosana[J]. Food Chemistry, 2014, 145(5): 1 066-1 071.
[20] ZHANG C, WANG W, LU R L,et al.Metabolic responses of Beauveria bassiana to hydrogen peroxide-induced oxidative stress using an LC-MS-based metabolomics approach[J]. Journal of Invertebrate Pathology, 2016, 137: 1-9.
[21] HU F L, HE Y Q, HUANG B, et al. Secondary metabolites in a soybean fermentation broth of Paecilomyces militaris[J].Food Chemistry, 2009, 116(1): 198-201.
[22] 章能胜,王金彬,胡丰林,等.玫烟色拟青霉中过氧麦角甾醇的高速逆流色谱法分离纯化及质谱分析[J].食品与发酵工业,2009,35(6):14-17.
[23] DABUR R,MITTAL A. Detection and qualitative analysis of fatty acid amides in the urine of alcoholics using HPLC-QTOF-MS[J]. Alcohol, 2016, 52: 71-78.
[24] YANG Z Z, LI Z H, LI Z. DPPH-HPLC-MS assisted rapid identification of endothelial protective substances from Xiao-Ke-An[J].Journal of Ethnopharmacology,2017,211(7): 188-196.
