Production of extracellular enzymes by different isolates of Pochonia chlamydosporia

Ivânia Esteves , Belkis Peteira , Simon D. Atkins , Naresh Magan , Brian Kerry
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引用次数: 71

Abstract

For the first time, the specific activities of chitinases, esterases, lipases and a serine protease (VCP1) produced by different isolates of the nematophagous fungus Pochonia chlamydosporia were quantified and compared. The isolates were grown for different time periods in a minimal liquid medium or media supplemented with 1 % chitin, 0.2 % gelatin or 2 % olive oil. Enzyme-specific activities were quantified in filtered culture supernatants using chromogenic p-nitrophenyl substrates (for chitinases, lipases and esterases) and a p-nitroanilide substrate (to measure the activity of the proteinase VCP1). Additionally, information on parasitic growth (nematode egg parasitism) and saprotrophic growth (plant rhizosphere colonisation) was collected. Results showed that the production of extracellular enzymes was influenced by the type of medium (p < 0.05) in which P. chlamydosporia was grown. Enzyme activity differed with time (p < 0.05), and significant differences were found between isolates (p < 0.001) and the amounts of enzymes produced (p < 0.001). However, no significant relationships were found between enzyme activities and parasitic or saprotrophic growth using Kendall's coefficient of concordance or Spearman rank correlation coefficient. The results provided new information about enzyme production in P. chlamydosporia and suggested that the mechanisms which regulate the trophic switch in this fungus are complex and dependent on several factors.

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不同分离株衣孢波氏菌胞外酶的生产
首次定量比较了不同菌种的几丁质酶、酯酶、脂肪酶和丝氨酸蛋白酶(VCP1)的比活性。分离株在少量液体培养基或添加1%甲壳素、0.2%明胶或2%橄榄油的培养基中生长不同时间。用显色对硝基苯基底物(几丁质酶、脂肪酶和酯酶)和对硝基苯胺底物(测定蛋白酶VCP1的活性)在过滤后的培养上清中定量测定酶的特异性活性。此外,还收集了寄生生长(线虫卵寄生)和腐养生长(植物根际定植)的信息。结果表明,胞外酶的产生受培养基类型的影响(p <0.05),其中衣孢假单胞菌生长。酶活性随时间不同而不同(p <0.05),分离株间差异有统计学意义(p <0.001)和酶的产生量(p <0.001)。然而,利用肯德尔一致性系数和斯皮尔曼秩相关系数,酶活性与寄生和腐养生长之间没有显著的相关关系。这些结果为衣孢单胞菌产酶提供了新的信息,并表明该真菌营养转换的调节机制是复杂的,依赖于几个因素。
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