红花醇提物特异性抑制酿酒酵母细胞壁合成研究
|
摘要
基于酿酒酵母及其葡聚糖合酶缺陷突变株组成的靶标定向全细胞筛选结果显示,葡聚糖合酶缺陷酵母Δfks1对中草药红花的醇提物更加敏感,其最低抑制质量浓度(MIC)值比野生型酵母Whu2a低8倍,红花醇提物对Δfks1的抑制作用能在山梨醇等渗溶液中得到部分挽救,酵母细胞在红花醇提物处理后β-葡聚糖含量降低了56.8%.推断红花醇提物中含有抑制真菌细胞壁葡聚糖合成的有效成分.
A whole-cell screening pattern consisting of wild-type Saccharomyces cerevisiae Whu2 a and glucan synthase-defective strain S.cerevisiae Δfks1 was used to screen against ethanol extracts of Carthamus tinctorius L.The result shows S.cerevisiae Δfks1 is more sensitive to the ethanol extract of herbal medicine C.tinctorius L.,its minimum inhibitory concentration(MIC)value is 8 times lower than S.cerevisiae Whu2 a,and the inhibition effect against S.cerevisiae Δfks1 can be partly rescued by sorbitol.After the treatment by the ethanol extract of C.tinctorius L.,the content ofβ-glucan on the cell wall decreased by 56.8%,indicating that there are active substances against the glucan synthesis of fungal cell wall.The purification and identification of the target substances will provide experimental techniques for developing new antifungal drugs.
A whole-cell screening pattern consisting of wild-type Saccharomyces cerevisiae Whu2 a and glucan synthase-defective strain S.cerevisiae Δfks1 was used to screen against ethanol extracts of Carthamus tinctorius L.The result shows S.cerevisiae Δfks1 is more sensitive to the ethanol extract of herbal medicine C.tinctorius L.,its minimum inhibitory concentration(MIC)value is 8 times lower than S.cerevisiae Whu2 a,and the inhibition effect against S.cerevisiae Δfks1 can be partly rescued by sorbitol.After the treatment by the ethanol extract of C.tinctorius L.,the content ofβ-glucan on the cell wall decreased by 56.8%,indicating that there are active substances against the glucan synthesis of fungal cell wall.The purification and identification of the target substances will provide experimental techniques for developing new antifungal drugs.
引文
[1]ZHANGL L,TIAN K,TANG Z H,et al.Phytochemistry and pharmacology of Carthamus tinctorius L[J].The american journal of chinese medicine,2016,44(2):197-226.
[2]白亮亮,邹鸣峰,王恒,等.3种红花的体外抑菌作用研究[J].食品科技,2015(2):311-314.
[3]汪倩雯,汪琨,杨霄,等.89种中草药醇提物体外抗菌活性的研究[J].浙江工业大学学报,2009,37(3):289-294.
[4]崔志峰,王燕华,马宁,等.黄连醇提物对真菌几丁质合酶的抑制作用[J].浙江工业大学学报,2014,42(1):16-19.
[5]LIU M,SEIDEL V,KATERERE D R,et al.Colorimetric broth microdilution method for the antifungal screening of plant extracts against yeasts[J].Methods,2007,42:325-329.
[6]CUI Z,DING Z,YANG X,et al.Gene disruption and characterization of a class V chitin synthase in Botrytis cinerea[J].Canadian journal of microbiology,2009,55(11):1267-1274.
[7]PEIXOTO L R,ROSALEN P L,FERREIRA G L S,et al.Antifungal activity,mode of action and anti-biofilm effects of Laurus nobilis Linnaeuses sential oil against Candidaspp[J].Archives of oral biology,2017,73:179-185.
[8]PFALLER M,RILEY J,KOERNER T.Effects of cilofungin(LY121019)on carbohydrate and sterol composition of Candida albicans[J].European journal of clinical microbiology and infectious diseases,1989(12):1067-1070.
[9]HARTL L,ZACH S,SEIDL-SEIBOTH V.Fungal chitinases:diversity,mechanistic properties and biotechnological potential[J].Applied microbiology and biotechnology,2012,93(2):533-543.
[10]CRAGG G M,NEWMAN D J.Natural products:a continuing source of novel drug leads[J].Biochimica et biophysica acta(BBA)-general subjects,2013,1830(6):3670-3695.
[11]YIM N H,HWANG E I,YUN B S,et al.Sesquiterpene furan compound CJ-01,a novel chitin synthase 2inhibitor from Chloranthus japonicus SIEB[J].Biological and pharmaceutical bulletin,2008,31(5):1041-1044.
[12]HWANG E I,LEE Y M,LEE S M,et al.Inhibition of chitin synthase 2and antifungal activity of lignans from the stem bark of Lindera erythrocarpa[J].Planta medica,2007,73(7):679-682.
[13]KANG T H,HWANG E I,YUN B S,et al.Chitin synthase2inhibitory activity of O-methyl pisiferic acid and 8,20-dihydroxy-9(11),13-abietadien-12-one,isolated fromChamaecyparis pisifera[J].Biological and pharmaceutical bulletin,2008,31(4):755-759.
[14]汪琨,徐峥,汪倩雯,等.肉桂醛特异性抑制酵母细胞壁合成的作用机理[J].食品与发酵工业,2012,38(3):68-72.
[15]HWANG E I,KWON B M,LEE S H,et al.Obovatols,new chitin synthase 2inhibitors of Saccharomyces cerevisiae from Magnolia obovata[J].Journal of antimicrobial chemotherapy,2002,49:95-101.
[16]LESAGE G,SDICU A M,MENARD P,et al.Analysis ofβ-1,3-glucan assembly in Saccharomyces cerevisiae using a synthetic interaction network and altered sensitivity to caspofungin[J].Genetics,2004,167(1):35-49.
[17]MONCIARDINI P,IORIO M,MAFFIOLI S,et al.Discovering new bioactive molecules from microbial sources[J].Microbial biotechnology,2014,7(3):209-220.
[18]刘玉庆,李晔,车程川,等.大肠杆菌对中草药敏感性试验及其方法研究[J].中兽医医药杂志,2003,22(1):3-5.
[19]FROST D J,BRANDT K D,CUGIER D,et al.Whole-cell Candida albicans assay for the detection of inhibitors towards fungal cell wall synthesis and assembly[J].Journal of antibiotics,1995,48(4):306-310.
[2]白亮亮,邹鸣峰,王恒,等.3种红花的体外抑菌作用研究[J].食品科技,2015(2):311-314.
[3]汪倩雯,汪琨,杨霄,等.89种中草药醇提物体外抗菌活性的研究[J].浙江工业大学学报,2009,37(3):289-294.
[4]崔志峰,王燕华,马宁,等.黄连醇提物对真菌几丁质合酶的抑制作用[J].浙江工业大学学报,2014,42(1):16-19.
[5]LIU M,SEIDEL V,KATERERE D R,et al.Colorimetric broth microdilution method for the antifungal screening of plant extracts against yeasts[J].Methods,2007,42:325-329.
[6]CUI Z,DING Z,YANG X,et al.Gene disruption and characterization of a class V chitin synthase in Botrytis cinerea[J].Canadian journal of microbiology,2009,55(11):1267-1274.
[7]PEIXOTO L R,ROSALEN P L,FERREIRA G L S,et al.Antifungal activity,mode of action and anti-biofilm effects of Laurus nobilis Linnaeuses sential oil against Candidaspp[J].Archives of oral biology,2017,73:179-185.
[8]PFALLER M,RILEY J,KOERNER T.Effects of cilofungin(LY121019)on carbohydrate and sterol composition of Candida albicans[J].European journal of clinical microbiology and infectious diseases,1989(12):1067-1070.
[9]HARTL L,ZACH S,SEIDL-SEIBOTH V.Fungal chitinases:diversity,mechanistic properties and biotechnological potential[J].Applied microbiology and biotechnology,2012,93(2):533-543.
[10]CRAGG G M,NEWMAN D J.Natural products:a continuing source of novel drug leads[J].Biochimica et biophysica acta(BBA)-general subjects,2013,1830(6):3670-3695.
[11]YIM N H,HWANG E I,YUN B S,et al.Sesquiterpene furan compound CJ-01,a novel chitin synthase 2inhibitor from Chloranthus japonicus SIEB[J].Biological and pharmaceutical bulletin,2008,31(5):1041-1044.
[12]HWANG E I,LEE Y M,LEE S M,et al.Inhibition of chitin synthase 2and antifungal activity of lignans from the stem bark of Lindera erythrocarpa[J].Planta medica,2007,73(7):679-682.
[13]KANG T H,HWANG E I,YUN B S,et al.Chitin synthase2inhibitory activity of O-methyl pisiferic acid and 8,20-dihydroxy-9(11),13-abietadien-12-one,isolated fromChamaecyparis pisifera[J].Biological and pharmaceutical bulletin,2008,31(4):755-759.
[14]汪琨,徐峥,汪倩雯,等.肉桂醛特异性抑制酵母细胞壁合成的作用机理[J].食品与发酵工业,2012,38(3):68-72.
[15]HWANG E I,KWON B M,LEE S H,et al.Obovatols,new chitin synthase 2inhibitors of Saccharomyces cerevisiae from Magnolia obovata[J].Journal of antimicrobial chemotherapy,2002,49:95-101.
[16]LESAGE G,SDICU A M,MENARD P,et al.Analysis ofβ-1,3-glucan assembly in Saccharomyces cerevisiae using a synthetic interaction network and altered sensitivity to caspofungin[J].Genetics,2004,167(1):35-49.
[17]MONCIARDINI P,IORIO M,MAFFIOLI S,et al.Discovering new bioactive molecules from microbial sources[J].Microbial biotechnology,2014,7(3):209-220.
[18]刘玉庆,李晔,车程川,等.大肠杆菌对中草药敏感性试验及其方法研究[J].中兽医医药杂志,2003,22(1):3-5.
[19]FROST D J,BRANDT K D,CUGIER D,et al.Whole-cell Candida albicans assay for the detection of inhibitors towards fungal cell wall synthesis and assembly[J].Journal of antibiotics,1995,48(4):306-310.