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Enzymatic Characterization of Three Entomopathogenic Fungi with the API ZYM System
以API ZYM系統測定蟲生真菌之酵素特性
摘要
The enzymatic profiles of Beauveria bassiana (Bb), Metarihizium anisopliae (Ma), Verticillium lecanii (VI), and their subcultures were studied by using the API ZYM system. Isolates all showed no reaction to substrates with β-glucuronidase, or cystein aminopeptidase except for Ma (str. 195). All isolates showed little activity or were completely without activity to substrates with lipase, trysin, chymotrysin, α-glucosidase, or α-fucosidase. However, Bb Ho 42 reacted strongly to substrate with α-mannosidase. Both isolates of M. anisopliae (strs. 194 and 195) reacted strongly to substrates with leucine aminopeptidase, acid phosphatase, phosphoamidase, or β-glucosaminidase. Additionally, M. anisopliae str. 194 reacted strongly to the substrates with esterase (C_4), or esterase lipase, while M. anisoplaie str. 195 moderately reacted to the substrates with β-glucosidase. Results showed that the enzymatic activity of B. bassiana against substrates with phosphatase alkaline, leucine aminopeptidase, α-galactodase, or α-galactodase declined after serial passages from subcultures of Ho. 212 F_3 to F_9. V. lecanii exhibited the tendency of declining activity against substrates with leucine aminopeptidase, valine aminopeptidase, phosphosamidase, β-glucosidase, orβ-glucosaminidase from subcultures of F_4 to F_6. On the other hand, the reaction to the substrate with phosphatase alkline increased. The results of this study indicate that API ZYM techniques provide a rapid and reproducible semi-quantitative method for revealing enzymatic activity.
並列摘要
本實驗利用半定量之API ZYM系統來研究三種蟲生真菌，白殭菌Beauveria bassiana，黑殭菌Metarhizium anisopliae，蠟蚧幹枝孢菌Verticillium lecanii的酵素剖圖。所有的分離株對β-glucuronidase及cysteine aminopeptidase的基質無反應，但黑殭菌195菌株對後者有反應。所有的分離株對Lipase, trypsin, chymotrypsin, α-glucos idase及α-fucosidase的基質無反應或只有輕微的反應。而白殭菌Col 42品系對α-mann osidase基質有很強的反應。黑殭菌的二個分離株(194及195)對leucine aminopeptidase, phosphatase acid, phosphoamidase, β-glucosaminidase具有強烈之反應。除此外，黑殭菌194對esterase, esterase lipase和β-glucosidase的基質反應較強而黑殭菌195則只有中度的反應。就繼代培養而言，白殭菌第九代之培養對phosphatase alkaline, leucine aminopeptidase, α-galatosidase, β-galactosidase的基質反應活性會下降。蠟蚧幹枝孢菌第六代之培養對leucine aminopeptidase, valine aminopeptidase, phosphoamidase, β-glucosidase，及β-glucosaminidase反應活性會下降。相反的對phosphatase alkaline反應有些微上升。以上的結果顯示API ZYM測試方法可提供快速的半定量的方法來辨識不同分離株及繼代培養的酵素活性。