采用平板法与斜面纯化法从河南红油香椿中分离内生真菌,并采用紫外分光光度法测定其次级代谢产物的α-糖苷酶抑制活性及抗氧化活性,牛津杯法测定其抗细菌活性,采用分子生物学方法鉴定高活性菌株。从香椿中共分离得到内生真菌6株,其中,菌株56-50的次级代谢产物具有最高α-糖苷酶抑制活性,为(24.98±1.89)%。菌株56-50和TS47均具有较好的抗氧化活性,其中,菌株56-50对ABTS+·和DPPH·的清除活性分别为(95.92±0.40)%和(91.77±0.45)%,TS47对DPPH·的清除活性为(94.55±0.15)%。菌株56-50和TS8分别对豪氏变形杆菌和欧文氏菌具有较强抑制活性,抑菌圈为(20.33±0.08)mm和(15.11±0.07)mm。综上,菌株56-50均具有较好的α-糖苷酶抑制活性、抗氧化和抗细菌活性,并根据18S rDNA测序结果将其鉴定为链格孢属。该研究为利用植物内生真菌发酵生产α-糖苷酶抑制剂、天然抗氧化剂和抗菌剂提供理论依据。
The endophytic fungi from fresh ‘Hongyou’ Chinese toon (Toona sinensis) in Henan province were isolated and purified, their α-glucosidase inhibitory, antibacterial and antioxidative activities were determined, and the species of the active strain was identified. A total of six endophytic fungi were isolated, and the secondary metabolites of strain 56-50 had the highest α-glucosidase inhibitory activity, which was (24.98±1.89%). Both strains 56-50 and TS47 had good antioxidative activities, as the scavenging effects of strain 56-50 on ABTS+· and DPPH· were (95.92±0.40)% and (91.77±0.45)%, respectively, while TS47 could remove (94.55±0.15)% DPPH· . Moreover, strains 56-50 and TS8 had strong inhibitory activities against Proteus hauseri and Erwinia sp., respectively, with inhibition zones of (20.33±0.08) and (15.11±0.07) mm, respectively. Strain 56-50, that exhibited strong α-glucosidase inhibitory activity, antioxidant activity and antibacterial activity, was identified as Alternaria sp. according to 18S rDNA sequencing. This study provides a theoretical basis for producing α-glucosidase inhibitors, natural antioxidants and antibacterial agents by using plant endophytic fungi.
[1] 王赵改, 陈丽娟,张乐,等. 不同采收期红油香椿营养成分和抗氧化活性分析[J]. 食品科学, 2015, 36(4): 158-163.
[2] MENG Q Q, PENG X R, LU S Y, et al. Lactam triterpenoids from the bark of Toona sinensis[J]. Natural Products and Bioprospecting, 2016, 6(5): 239-245.
[3] WANG C Y, LIN K H, YANG C H, et al. Toona sinensis extracts induced cell cycle arrest and apoptosis in the human lung large cell carcinoma[J]. The Kaohsiung Journal of Medical Sciences, 2010, 26(2): 68-75.
[4] ZHAO J, ZHOU X W, CHEN X B, et al. α-Glucosidase inhibitory constituents from Toona sinensis[J]. Chemistry of Natural Compounds, 2009, 45(2): 244-246.
[5] 任美萍, 李春红, 李蓉. 香椿总黄酮对糖尿病小鼠及正常小鼠血糖的影响[J]. 泸州医学院学报, 2012, 35(3): 261-262.
[6] ZHANG W, LI C, YOU L J, et al. Structural identification of compounds from Toona sinensis leaves with antioxidant and anticancer activities[J]. Journal of functional foods, 2014,10: 427-435.
[7] HSEU Y C, CHANG W H, CHEN C S, et al. Antioxidant activities of Toona Sinensis leaves extracts using different antioxidant models[J]. Food and Chemical Toxicology, 2008, 46(1): 105-114.
[8] WU J G, PENG W, YI J, et al. Chemical composition, antimicrobial activity against Staphylococcus aureus and a pro-apoptotic effect in SGC-7901 of the essential oil from Toona sinensis (A.Juss.) Roem. leaves[J]. Journal of Ethnopharmacology, 2014, 154(1), 198-205.
[9] 陈丛瑾. 香椿有效成分的提取纯化和生物活性研究[D]. 长沙:中南林业科技大学, 2010: 168-172.
[10] LI X, WANG J, ZHANG S, et al. Distribution of fungal endophytes in roots of Stipa krylovii across six vegetation types in grassland of northern China[J]. Fungal Ecology, 2018, 31: 47-53.
[11] 曾松荣, 徐成东,王海坤,等. 药用植物内生真菌及其具宿主相同活性成分的机制初探[J]. 中草药, 2000, 31(4): 306-308.
[12] 赵凯,宇璐,金昱言,等.内生真菌紫衫醇生物合成的研究现状与展望[J].生物工程学报, 2016, 32(8): 1 038-1 051.
[13] KIM Y M, WANG M H, RHEE H I. A novel a-glucosidase inhibitor from pine bark[J]. Carbohydrate Research, 2004, 339(3): 715-717.
[14] 马燕燕, 鲁晓翔. 天然产物α-葡萄糖苷酶抑制剂筛选研究进展[J]. 粮食与油脂, 2010, 170(6): 7-10.
[15] CHEN C, YOU L J, ABBASI A M, et al. Optimization for ultrasound extraction of polysaccharides from mulberry fruits with antioxidant and hyperglycemic activity in vitro[J]. Carbohydrate Polymers, 2015, 130(5): 122-132.
[16] GUO L D, HYDE K D, LIEW E C Y. Identification of endophytic fungi from Livistona chinensis based on morphology and rDNA sequences[J]. New Phytologist, 2000, 147(3): 617-630.
[17] 赵文竹. 玉米须功能因子活性评价及其降血糖机理研究[D]. 长春:吉林大学, 2014: 63-64.
[18] OZCAN E.A novel autmated direct measurement method for total antioxidant capacity using a new generation,more stable ABTS radical cation[J].Clinical Biochemistry,2004,37(4):277-285.
[19] 赵呈雷. 芦蒿秸秆提取物抗氧化作用及其制剂学初步研究[D]. 镇江:江苏大学, 2007: 37-38.
[20] SUN T, HO C T. Antioxidant activities of buckwheat extracts[J]. Food Chemistry, 2005, 90 (4): 743-749.
[21] GOMEZ S, COSSON C, DESCHAMPS A M. Evidence for a bacteriocin-like substance produced by a new strain of Streptococcus sp., inhibitory to Gram-positive food-borne pathogens [J]. Research in Microbiology, 1997, 148(9): 757-766.
[22] 王晓敏, 王惠, 刘天行,等. 一株不产生孢子的盐生海芦笋内生真菌鉴定[J]. 食品科学, 2013, 34(17): 146-149.
[23] 谌小立, 陈文莹, 夏吉跃,等. 红酸汤烹调过程中全反式番茄红素含量变化规律研究[J]. 中国调味品, 2018, 43(2): 68-71.
[24] 谌小立, 黄先静, 杨旭芹,等. 三种食用油对红酸汤烹调中全反式番茄红素含量影响研究[J]. 中国调味品, 2019, 44(3): 91-94.
[25] 郑红梅, 叶耀辉,王婷,等. 药用植物内生真菌及其代谢产物活性的研究概况[J]. 江西中医药, 2016, 47(4): 71-73.
[26] STIERLE A, STROBEL G, STIERLE D. Taxol and taxane production by Taxomyces andreanae, an endophytic fungus of Pacific yew [J]. Science, 1993, 260(5 105): 214-216.
[27] 周生亮, 陈双林. 薯蓣内生真菌抗氧化活性的初步研究[J]. 微生物学杂志, 2006,26(1): 40-43.
[28] 刘军生, 解修超, 罗阳兰,等. Aspergillus sp. EA-LJS80的分离及其发酵液提取物七叶皂苷C的生物活性[J]. 菌物学报, 2018,37(1): 79-87.
