PURIFICATION, CHARACTERIZATION, AND SYNERGISTIC ACTIVITY OF A GLUCAN 1,3-BETA-GLUCOSIDASE AND AN N-ACETYL-BETA-GLUCOSAMINIDASE FROM TRICHODERMA-HARZIANUM
PURIFICATION, CHARACTERIZATION, AND SYNERGISTIC ACTIVITY OF A GLUCAN 1,3-BETA-GLUCOSIDASE AND AN N-ACETYL-BETA-GLUCOSAMINIDASE FROM TRICHODERMA-HARZIANUM
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DOI：
10.1094/phyto-84-398
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发表时间：
1994-04-01
期刊：
影响因子：
3.2
通讯作者：
HARMAN, GE
中科院分区：
文献类型：
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作者：
LORITO, M;HAYES, CK;HARMAN, GE
从哈茨木霉菌株 P1 的培养滤液中将葡聚糖 1,3-β-葡萄糖苷酶 (EC 3.2.1.58) 和 N-乙酰基-β-氨基葡萄糖苷酶 (EC 3.2.1.30) 纯化至同质。葡聚糖1,3-β-葡萄糖苷酶的分子量和PI分别为78 kDa和6.2，N-乙酰基-β-氨基葡萄糖苷酶的分子量和PI分别为72 kDa和4.6。针对灰葡萄孢测试了葡聚糖 1,3-β-葡萄糖苷酶和 N-乙酰基-β-氨基葡萄糖苷酶，并将它们的抗真菌活性与同样从 T. harzianum 菌株 P1 纯化的内切壳多糖酶和几丁质 1,4-β-壳二糖苷酶进行了比较。四种细胞壁降解酶也作为包含两种、三种或所有四种蛋白质的所有可能组合的混合物进行了测试。当同时使用两种、三种或四种酶时，检测到对灰霉病菌的孢子萌发和芽管伸长具有协同抑制作用。当使用含有四种不同细胞壁降解酶的溶液时，获得了最高水平的抗真菌活性。抑制分生孢子萌发的ED(50)（50%有效剂量）值低至1.6μgml(-1)，抑制存活孢子芽管伸长的ED(50)值低至1.7μgml(-1)。
A glucan 1,3-beta-glucosidase (EC 3.2.1.58) and an N-acetyl-beta-glucosaminidase (EC 3.2.1.30) were purified to homogeneity from the culture filtrate of Trichoderma harzianum strain P1. The molecular masses and the pis were 78 kDa and 6.2, respectively, for the glucan 1,3-beta-glucosidase and 72 kDa and 4.6, respectively, for the N-acetyl-beta-glucosaminidase. The glucan 1,3-beta-glucosidase and the N-acetyl-beta-glucosaminidase were tested against Botrytis cinerea, and their antifungal activity was compared with that obtained for an endochitinase and a chitin 1,4-beta-chitobiosidase also purified from T. harzianum strain P1. The four cell wall-degrading enzymes were also tested as mixtures containing two, three, or all four proteins in all possible combinations. A synergistic, inhibitory effect was detected on both spore germination and germ tube elongation of B. cinerea when two, three, or four enzymes were applied together. The highest level of antifungal activity was obtained when a solution containing four different cell wall-degrading enzymes was used. ED(50) (50% effective dose) values were as low as 1.6 mu g ml(-1) for inhibition of conidial germination and 1.7 mu g ml(-1) for inhibition of germ tube elongation of the surviving spores.