柠檬皮中柠檬苦素对黑曲霉的抑菌特性和机理
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摘要
本研究以柠檬皮中柠檬苦素为原料,对其抑制黑曲霉的抑菌活性、抑菌活性的稳定性和抑菌机理进行了研究。结果表明:柠檬皮中柠檬苦素对黑曲霉的最小抑菌浓度(MIC)和最小杀菌浓度(MFC)分别为625μg/mL、5000μg/mL;对孢子萌发和菌丝生长抑制的半最大效应浓度(EC50)分别为311.03μg/mL和291.63μg/mL。柠檬苦素对黑曲霉的抑菌活性在紫外光照下受到抑制;在pH值为6～7时受到抑制;金属离子Fe3+、Fe2+可增强其抑菌活性。在柠檬皮中柠檬苦素作用下,黑曲霉的胞外碱性磷酸酶(AKP)活力、可溶性蛋白和菌丝体总糖含量发生很大变化;扫描电子显微镜观察发现黑曲霉孢子发生了变形、破裂、溶出物附着的现象。因此,柠檬苦素破坏了黑曲霉细胞壁和细胞膜的完整性,造成大量内容物渗出,影响了细胞正常代谢,进而抑制了菌体细胞的生长繁殖。
The antifungal activity,stability and mechanism of limonin from lemon peel against Aspergillus niger were investigated.The results showed that the minimal inhibitory concentration(MIC) and the minimal fungicidal concentration(MFC) of limonin from lemon peel on Aspergillus Niger were 625 μg/mL and 5000 μg/mL respectively,and the half-effective concentration(EC50) on spore germination and mycelium growth were 311.03 μg/mL and 291.63 μg/mL respectively.The antifungal activity of limonin on Aspergillus niger was weakened under UV light and pH 6～7.Fe3+ and Fe2+ could enhance the antifungal activity.The limonin from lemon peel could greatly change extracellular alkaline phosphatase(AKP) activity,soluble protein and total sugar content of mycelium of Aspergillus niger.Therefore,limonin destroyed the integrity of cell wall and cell membrane of Aspergillus niger,resulting in a large amount of content exudation,affecting the normal metabolism of cells,thereby inhibiting the growth and reproduction of cells.
The antifungal activity,stability and mechanism of limonin from lemon peel against Aspergillus niger were investigated.The results showed that the minimal inhibitory concentration(MIC) and the minimal fungicidal concentration(MFC) of limonin from lemon peel on Aspergillus Niger were 625 μg/mL and 5000 μg/mL respectively,and the half-effective concentration(EC50) on spore germination and mycelium growth were 311.03 μg/mL and 291.63 μg/mL respectively.The antifungal activity of limonin on Aspergillus niger was weakened under UV light and pH 6～7.Fe3+ and Fe2+ could enhance the antifungal activity.The limonin from lemon peel could greatly change extracellular alkaline phosphatase(AKP) activity,soluble protein and total sugar content of mycelium of Aspergillus niger.Therefore,limonin destroyed the integrity of cell wall and cell membrane of Aspergillus niger,resulting in a large amount of content exudation,affecting the normal metabolism of cells,thereby inhibiting the growth and reproduction of cells.
引文
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[14]徐亚超,盛杰,刘安军.白酒发酵副产物黄水抑菌特性及稳定性研究[J].食品科学,2017,38(15):122-126XU Ya-Chao,CHENG Jie,LIU An-jun.Antimicrobial properties and stability of yellow water,a byproduct of Chinese liquor fermentation[J].Food Science,2017,38(15):122-126
[15]许女,史改玲,张浩,等.植物乳杆菌KF1对奶牛乳房炎金黄色葡萄球菌的抑菌机制[J].中国食品学报,2016,16(10):19-27XU Nyu,SHI Gai-ling,ZHANG Hao,et al.Antibacterial mechanism of lactobacillus plantarum KF1 on Staphylococcus aureus isolated from subclinical mastitis milk[J].Journal of Chinese Institute of Food Science and Technology,2016,16(10):19-27
[16]何梦影.基于脂质体模型对黄酮类化合物抑菌机理的研究[D].华中农业大学,2014HE Meng-yin.Antimicrobial mechanism of flavonoids by model membrane study[D].Huazhong Agricultural University,2014
[2]晏敏,周宇,贺肖寒,等.柑橘籽中柠檬苦素及类似物的生物活性研究进展[J].食品与发酵工业,2018,44(2):290-296YAN Min,ZHOU Yu,HE Xiao-han,et al.Research progress on the bioactivity of limonin and analogues in Citrus[J].Food and Fermentation Industries,2018,44(2):290-296
[3]李科静.大米中霉菌的微波杀菌工艺及机理研究[D].吉林:吉林大学,2015LI Ke-jing.Studies on microwave sterilization process and mechanism of molds in rice[D].Jilin:Jilin University,2015
[4]李长平,杨俊,黄河,等.源于凡纳滨对虾血蓝蛋白化学合成肽段的抗黑曲霉活性[J].中国水产科学,2018,25(1):189-194LI Chang-ping,YANG Jun,HUANG He,et al.Anti-aspergillus niger activities on the chemical synthesis peptides derived from hemocyanin in shrimp Litopenaeus vannamei[J].Journal of Fishery Sciences of China,2018,25(1):189-194
[5]马欢欢,林洋,吕欣然,等.96孔板法筛选抗黑曲霉性乳酸菌及抑菌机理研究[J].食品工业科技,2017,38(12):171-175MA Huan-huan,LIN Yang,LYU Xin-ran,et al.Screening and inhibition mechanism of lactic acid bacteria against Aspergillus niger using 96-well microtiter plates[J].Science and Technology of Food Industry,2017,38(12):171-175
[6]李彪,施蕊,熊智,等.柚子皮中柠檬苦素提取工艺及其抑菌活性研究[J].食品工业科技,2012,33(5):198-200LI Biao,SHI Rui,XIONG Zhi,et al.Study on extraction and antibacterial activities of limonin in peel of Citrus maxima[J].Science and Technology of Food Industry,2012,33(5):198-200
[7]Amit Vikram,Guddadarangavvahally K,Jayaprakasha,Palmy R,et al.Limonin 7-methoxime interferes with Escherichia coli biofilm formation and attachment in type 1pili and antigen 43 dependent manner[J].Food Control,2012,26(2)
[8]周梦娇,万春鹏,陈金印.柑橘绿霉病中草药高效抑菌剂的筛选及抑菌机理研究[J].现代食品科技,2014,30(3):144-149,75ZHOU Meng-jiao,WAN Chun-peng,CHEN Jin-yin.Screening and antifungal mechanism of Chinese herb extracts against green mold of Citrus[J].Modern Food Science and Technology,2014,30(3):144-149,75
[9]Qiu M,Wu C,Ren G,et al.Effect of chitosan and its derivatives as antifungal and preservative agents on postharvest green Asparagus[J].Food Chemistry,2014,155(15):105-111
[10]蓝蔚青,车旭,谢晶,等.复合生物保鲜剂对荧光假单胞菌的抑菌活性及作用机理[J].中国食品学报,2016,16(8):159-165LAN Wei-qing,CHE Xu,XIE Jing,et al.Antibacterial activity and mechanism of composite biological preservatives against Pseudomonas fluorescens[J].Journal of Chinese Institute of Food Science and Technology,2016,16(8):159-165
[11]任先伟,魏晓璐,黄鑫,等.核桃青皮提取物抑菌活性及抑菌机理研究[J].食品工业科技,2015,36(18):93-98REN Xian-wei,WEI Xiao-lu,HUANG Xin,et al.Antibacterial activity and mechanism of walnut green husk’extract[J].Science and Technology of Food Industry,2015,36(18):93-98
[12]韩亚珊.食品化学实验指导[M].北京:中国农业出版社,1996HAN Ya-shan.Experimental Guide for Food Chemistry[M].Beijing:China Agriculture Press,1996
[13]张传军,刘超,姜晓坤.薤白乙醇提取物的抑菌特性[J].食品科学,2011,32(5):119-122ZHANG Chuan-jun,LIU Chao,JIANG Xiao-kun.Antibacterial effect of ethanol extract from the bulbs of Allium macrostemon Bunge[J].Food Science,2011,32(5):119-122
[14]徐亚超,盛杰,刘安军.白酒发酵副产物黄水抑菌特性及稳定性研究[J].食品科学,2017,38(15):122-126XU Ya-Chao,CHENG Jie,LIU An-jun.Antimicrobial properties and stability of yellow water,a byproduct of Chinese liquor fermentation[J].Food Science,2017,38(15):122-126
[15]许女,史改玲,张浩,等.植物乳杆菌KF1对奶牛乳房炎金黄色葡萄球菌的抑菌机制[J].中国食品学报,2016,16(10):19-27XU Nyu,SHI Gai-ling,ZHANG Hao,et al.Antibacterial mechanism of lactobacillus plantarum KF1 on Staphylococcus aureus isolated from subclinical mastitis milk[J].Journal of Chinese Institute of Food Science and Technology,2016,16(10):19-27
[16]何梦影.基于脂质体模型对黄酮类化合物抑菌机理的研究[D].华中农业大学,2014HE Meng-yin.Antimicrobial mechanism of flavonoids by model membrane study[D].Huazhong Agricultural University,2014
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