抗真菌肽研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2020-0707

摘要/Abstract

摘要：

有害真菌已造成植物真菌病害、食物污染和人类真菌感染等问题,给人们的生活和生产带来极大危害,且其逐渐增加对抗真菌药物的耐药性,导致真菌防治日益困难。传统的合成类抗真菌药物具有药物残留和毒副作用,已不能满足需求,作为生物机体天然防御分子的抗真菌肽已成为应对真菌危害及耐药性的重要研究对象。抗真菌肽能够抑制有害真菌,具有高效、广谱、安全的特性,且其独特的抑菌机理解决了真菌耐药性的问题,在真菌防治中逐渐受到关注。基于此,主要从真菌的危害以及抗真菌肽的来源、分离纯化方法、抑菌机理等几个方面进行分析与论述,以期为抗真菌肽的研究提供参考。

关键词: 抗真菌肽, 真菌危害, 分离纯化, 抑菌机理

Abstract:

The harmful fungi have caused plant fungal disease,food contamination and human fungal infection,which can bring great harm to people’s life and production. Moreover,the increasing resistance of harmful fungi to antifungal drugs has made them difficult to control. Due to the drug residues and the toxic effects,the traditional synthetic fungicides have been unable to meet the people’s needs. Antifungal peptides,as the natural defense molecules of living organisms,have become an important research object in response to fungal hazards and drug resistance. Antifungal peptides can inhibit the harmful fungi,and have the characteristics of high efficiency,broad spectrum and high safety. The unique antifungal mechanism of antifungal peptides can solve the problem of fungal drug resistance,which has gradually attracted people’s attention in fungi control. Based on this,this paper mainly analyzed and discussed the following aspects such as the harm of fungi,the source of antifungal peptides,the method of isolation and purification,the antifungal mechanism of antifungal peptides,etc.,aiming to provide a reference for the study of antifungal peptides.

Key words: antifungal peptides, fungal hazard, isolation and purification, antifungal mechanism

王志新, 鲁雷震, 周景波, 封成玲, 贾紫伟, 宁亚维, 贾英民. 抗真菌肽研究进展[J]. 生物技术通报, 2021, 37(3): 206-218.

WANG Zhi-xin, LU Lei-zhen, ZHOU Jing-bo, FENG Cheng-ling, JIA Zi-wei, NING Ya-wei, JIA Ying-min. Advance in the Research of the Antifungal Peptides[J]. Biotechnology Bulletin, 2021, 37(3): 206-218.

图/表 5

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来源	前处理	初步分离	纯化	参考文献
蜘蛛	穿刺取血（放在柠檬酸钠缓冲液中,抗凝血）,离心去除血细胞,获得上清液	均质、酸化	反相高效液相色谱	[30]
海洋蜗牛	均质、离心,获得上清液	硫酸铵沉淀、离心、脱盐	反相高效液相色谱	[25]
天南星果	果肉清洗风干、乙醇溶液均质、离心,获得上清液	有机溶剂（丙酮）沉淀	反相高效液相色谱	[35]
蛹虫草	均质,离心,获得上清液	离子交换层析、凝胶过滤层析	快速蛋白质液相色谱	[40]
尼泊尔大红豆	蒸馏水均质,获得上清液	离子交换柱层析、亲和层析	快速蛋白质液相色谱	[37]
苦荞种子	磨碎,磷酸盐缓冲液浸提、离心,获得上清液	硫酸铵沉淀、透析	阴离子交换柱层析、琼脂糖凝胶柱层析、超滤	[34]
豆树种子	Tris-HCl溶液均质,离心,获得上清液	硫酸铵沉淀、透析	离子交换色谱（DEAE-纤维柱）、凝胶过滤色谱	[41]
乳酸菌IS10	发酵液离心,过滤除菌,获得上清液	上清液冷冻干燥	分子筛层析	[64]
芽孢杆菌BH072	离心、过滤除菌,获得滤液	硫酸铵沉淀	凝胶过滤色谱、D201阴离子交换树脂	[52]
内生芽孢杆菌B21	过滤,减压浓缩	有机溶剂（甲醇）萃取,硅胶柱层析	高效液相色谱	[7]
曲霉菌	过滤、离心,获得上清液,耐热性实验	热处理,离心,超滤	反相高效液相色谱	[45]
植物乳杆菌TE10	离心,过滤除菌,获得无细胞上清液	超滤	分子筛层析	[65]
枯草芽孢杆菌CP6	离心,获得上清液,100℃灭菌,冻干	甲醇抽提	离子交换层析、分子筛层析、反相高效液相色谱	[66]
枯草芽孢杆菌1.88株	离心,获得上清液	酸沉、乙醇抽提	反相高效液相色谱	[51]
短小芽孢杆菌HN-10	离心,获得上清液	硫氨酸沉淀、透析	AB-8大孔吸附树脂、Sephadex G-100凝胶、半制备型反相高效液相色谱	[54]
解淀粉芽孢杆菌BW-13	酸碱处理,离心,获得上清液	活性炭吸附	离子交换柱层析	[50]
丝状真菌SIIA-F1108	甲醇沉淀,过滤,获得滤液	大孔树脂吸附	正相硅胶色谱层析、高效液相色谱	[67]

来源	前处理	初步分离	纯化	参考文献
蜘蛛	穿刺取血（放在柠檬酸钠缓冲液中,抗凝血）,离心去除血细胞,获得上清液	均质、酸化	反相高效液相色谱	[30]
海洋蜗牛	均质、离心,获得上清液	硫酸铵沉淀、离心、脱盐	反相高效液相色谱	[25]
天南星果	果肉清洗风干、乙醇溶液均质、离心,获得上清液	有机溶剂（丙酮）沉淀	反相高效液相色谱	[35]
蛹虫草	均质,离心,获得上清液	离子交换层析、凝胶过滤层析	快速蛋白质液相色谱	[40]
尼泊尔大红豆	蒸馏水均质,获得上清液	离子交换柱层析、亲和层析	快速蛋白质液相色谱	[37]
苦荞种子	磨碎,磷酸盐缓冲液浸提、离心,获得上清液	硫酸铵沉淀、透析	阴离子交换柱层析、琼脂糖凝胶柱层析、超滤	[34]
豆树种子	Tris-HCl溶液均质,离心,获得上清液	硫酸铵沉淀、透析	离子交换色谱（DEAE-纤维柱）、凝胶过滤色谱	[41]
乳酸菌IS10	发酵液离心,过滤除菌,获得上清液	上清液冷冻干燥	分子筛层析	[64]
芽孢杆菌BH072	离心、过滤除菌,获得滤液	硫酸铵沉淀	凝胶过滤色谱、D201阴离子交换树脂	[52]
内生芽孢杆菌B21	过滤,减压浓缩	有机溶剂（甲醇）萃取,硅胶柱层析	高效液相色谱	[7]
曲霉菌	过滤、离心,获得上清液,耐热性实验	热处理,离心,超滤	反相高效液相色谱	[45]
植物乳杆菌TE10	离心,过滤除菌,获得无细胞上清液	超滤	分子筛层析	[65]
枯草芽孢杆菌CP6	离心,获得上清液,100℃灭菌,冻干	甲醇抽提	离子交换层析、分子筛层析、反相高效液相色谱	[66]
枯草芽孢杆菌1.88株	离心,获得上清液	酸沉、乙醇抽提	反相高效液相色谱	[51]
短小芽孢杆菌HN-10	离心,获得上清液	硫氨酸沉淀、透析	AB-8大孔吸附树脂、Sephadex G-100凝胶、半制备型反相高效液相色谱	[54]
解淀粉芽孢杆菌BW-13	酸碱处理,离心,获得上清液	活性炭吸附	离子交换柱层析	[50]
丝状真菌SIIA-F1108	甲醇沉淀,过滤,获得滤液	大孔树脂吸附	正相硅胶色谱层析、高效液相色谱	[67]

名称	来源	生物活性	分子质量/Da	参考文献
未命名	七鳃鳗血浆	抗青霉菌、曲霉菌	3 000	[24]
AcICK	蜜蜂体内	抗白僵菌、禾谷镰刀菌	6 600	[27]
LBLP	蜈蚣	抗白色念珠菌、假丝酵母等	2 757.6	[26]
Cm-p1	海洋蜗牛	抗红色毛癣菌、白色念珠菌、假丝酵母菌、灰霉菌、尖孢镰刀菌、黑曲霉等	1 485.26	[25]
Diapausin-1	烟草天蛾幼虫	抗酵母菌、黄色镰刀菌、禾谷镰刀菌等	5 000	[21]
Tn-AFP1	天南星果	抗热带假丝酵母菌	1 230	[35]
Cordymin	蛹虫草	抗玉米小斑病菌、花生球腔菌、立枯丝核菌、白色念珠菌菌丝生长、抗HIV-1逆转录酶和乳腺癌细胞增殖活性	10 906.62	[40]
Eryngin	食用菌杏鲍菇	抗尖孢镰刀菌、花生球腔菌菌丝生长	10 000	[46]
Skh-AMP1	木香叶片	抗曲霉和念珠菌属真菌	2 778.10	[38]
Pleurostrin	平菇	抗尖孢镰刀菌、花生球腔菌、苹果轮纹病菌菌丝生长	7 000	[47]
SmAMP3	鹰嘴豆草	抗腐皮镰孢菌、烟草赤星病菌、根腐离蠕孢、灰霉菌	3 363.9	[39]
未命名	苦荞种子	抗白腐菌、绿色木霉、链格孢霉	3 909	[33]
Campesin	甘蓝菜种子	抗尖孢镰刀菌、花生褐斑病菌,抗HepG2和MCF癌细胞的增殖	9 400	[32]
Ps-AFP1	植物种子	抗尖孢镰刀菌、白色念珠菌、黑曲霉、土曲霉	4 608.20	[68]
鞭毛蛋白	芽孢杆菌BH072	抗黑曲霉、尖孢镰刀菌、腐霉菌、灰霉菌	3 5615	[52]
W10	地衣芽孢杆菌	抗灰霉病菌,具有丝氨酸蛋白酶活性	48 794.16	[43]
AcAFP	曲霉菌	抗尖孢镰刀菌、腐皮镰孢菌、黑曲霉、灰霉菌、番茄早疫病菌	5 773	[45]
PcPAF	海洋桔青霉W1	抗绿色木霉、宛氏拟青霉、长柄链格孢、胶孢炭疽菌	约10 000	[69]
未命名	植物乳杆菌TE10	抗玉米种子黄曲霉MD3生长	664-2 024	[65]
未命名	芽孢杆菌AH-E-1	抗13种植物致病真菌和20种人类致病真菌	500-1 000	[6]
P-1	短小芽孢杆菌HN-10	抗粉红单端孢	1 149.14	[54]
未命名	枯草芽孢杆菌B25	抗尖孢镰刀菌、棒孢霉菌、茄链格孢菌、葡萄孢菌、胶孢炭疽菌、黑曲霉菌	38 708.67	[42]

名称	来源	生物活性	分子质量/Da	参考文献
未命名	七鳃鳗血浆	抗青霉菌、曲霉菌	3 000	[24]
AcICK	蜜蜂体内	抗白僵菌、禾谷镰刀菌	6 600	[27]
LBLP	蜈蚣	抗白色念珠菌、假丝酵母等	2 757.6	[26]
Cm-p1	海洋蜗牛	抗红色毛癣菌、白色念珠菌、假丝酵母菌、灰霉菌、尖孢镰刀菌、黑曲霉等	1 485.26	[25]
Diapausin-1	烟草天蛾幼虫	抗酵母菌、黄色镰刀菌、禾谷镰刀菌等	5 000	[21]
Tn-AFP1	天南星果	抗热带假丝酵母菌	1 230	[35]
Cordymin	蛹虫草	抗玉米小斑病菌、花生球腔菌、立枯丝核菌、白色念珠菌菌丝生长、抗HIV-1逆转录酶和乳腺癌细胞增殖活性	10 906.62	[40]
Eryngin	食用菌杏鲍菇	抗尖孢镰刀菌、花生球腔菌菌丝生长	10 000	[46]
Skh-AMP1	木香叶片	抗曲霉和念珠菌属真菌	2 778.10	[38]
Pleurostrin	平菇	抗尖孢镰刀菌、花生球腔菌、苹果轮纹病菌菌丝生长	7 000	[47]
SmAMP3	鹰嘴豆草	抗腐皮镰孢菌、烟草赤星病菌、根腐离蠕孢、灰霉菌	3 363.9	[39]
未命名	苦荞种子	抗白腐菌、绿色木霉、链格孢霉	3 909	[33]
Campesin	甘蓝菜种子	抗尖孢镰刀菌、花生褐斑病菌,抗HepG2和MCF癌细胞的增殖	9 400	[32]
Ps-AFP1	植物种子	抗尖孢镰刀菌、白色念珠菌、黑曲霉、土曲霉	4 608.20	[68]
鞭毛蛋白	芽孢杆菌BH072	抗黑曲霉、尖孢镰刀菌、腐霉菌、灰霉菌	3 5615	[52]
W10	地衣芽孢杆菌	抗灰霉病菌,具有丝氨酸蛋白酶活性	48 794.16	[43]
AcAFP	曲霉菌	抗尖孢镰刀菌、腐皮镰孢菌、黑曲霉、灰霉菌、番茄早疫病菌	5 773	[45]
PcPAF	海洋桔青霉W1	抗绿色木霉、宛氏拟青霉、长柄链格孢、胶孢炭疽菌	约10 000	[69]
未命名	植物乳杆菌TE10	抗玉米种子黄曲霉MD3生长	664-2 024	[65]
未命名	芽孢杆菌AH-E-1	抗13种植物致病真菌和20种人类致病真菌	500-1 000	[6]
P-1	短小芽孢杆菌HN-10	抗粉红单端孢	1 149.14	[54]
未命名	枯草芽孢杆菌B25	抗尖孢镰刀菌、棒孢霉菌、茄链格孢菌、葡萄孢菌、胶孢炭疽菌、黑曲霉菌	38 708.67	[42]