Kriaa Mouna, Hammami Inès, Sahnoun Mouna, Azebou Manel Cheffi, Triki Mohamed Ali, Kammoun Radhouane
Laboratoire de Microorganisme et de Biomolécules, Centre de Biotechnologie de Sfax, Université de Sfax, Route de Sidi Mansour Km 6, B.P. 1177, 3018, Sfax, Tunisia.
Ecole Supérieure d'Agriculture du Kef, Université de Jendouba, boulifa, 7119, kef, Tunisia.
Bioprocess Biosyst Eng. 2015 Nov;38(11):2155-66. doi: 10.1007/s00449-015-1455-y. Epub 2015 Aug 18.
This study was carried out to evaluate the in vitro and in vivo antifungal efficiency of Aspergillus tubingensis CTM 507 glucose oxidase (GOD) against plant pathogenic fungi. GOD displayed a wide inhibitory spectrum toward different fungi at a concentration of 20 AU. The GOD had a strong inhibitor effect on mycelia growth and spore germination of Pythium ultimum. Interestingly, the GOD exhibited a potent in vivo antifungal effect against P. ultimum responsible for potato plants disease. The antifungal GOD was purified 13-fold with 27 % yield and a specific activity of 3435 U/mg. The relative molecular mass of the GOD was 180 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The GOD activity was optimum at pH 4.5 and 60 °C. It was found to be stable over a large pH range (3-9). It also displayed a marked thermostability with a 50-min half-life at 65 °C. The 10 residues of the N-terminal sequence of the purified GOD (S-K-G-S-A-V-T-T-P-D) showed no homology to the other reported GOD, identifying a novel GOD. FTIR spectroscopic analysis revealed the presence of C-O and C=O groups corresponding to a D-glucono-lactone. The findings indicated that GOD is the first A. tubingensis-produced fungicide ever reported to exhibit such promising biological properties. It could become a natural alternative to synthetic fungicides to control certain important plant microbial diseases.
本研究旨在评估泡盛曲霉CTM 507葡萄糖氧化酶(GOD)对植物病原真菌的体外和体内抗真菌效果。GOD在浓度为20 AU时对不同真菌表现出广泛的抑制谱。GOD对终极腐霉的菌丝体生长和孢子萌发具有强烈的抑制作用。有趣的是,GOD对导致马铃薯植株病害的终极腐霉表现出强大的体内抗真菌效果。抗真菌GOD经纯化后比活提高了13倍,得率为27%,比活性为3435 U/mg。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)测定,GOD的相对分子质量为180 kDa。GOD的活性在pH 4.5和60℃时最佳。发现在较大的pH范围(3 - 9)内它是稳定的。在65℃时它还表现出显著的热稳定性,半衰期为50分钟。纯化的GOD的N端序列的10个残基(S-K-G-S-A-V-T-T-P-D)与其他已报道的GOD没有同源性,确定为一种新型GOD。傅里叶变换红外光谱分析揭示了对应于D-葡萄糖酸内酯的C-O和C=O基团的存在。研究结果表明,GOD是首次报道的具有如此有前景生物学特性的泡盛曲霉产生的杀真菌剂。它可能成为控制某些重要植物微生物病害的合成杀真菌剂的天然替代品。