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Analysis of Airborne Actinomycete Spores with Fluorogenic Substrates

机译:带有荧光底物的气载放线菌孢子的分析

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摘要

The reactions between seven fluorogenic substrates and different groups of enzymes, esterases, lipases, phosphatases, and dehydrogenases, were studied in a search for a new method for the detection of actinomycete spores. Fluorescence measurement was chosen as a fast and sensitive method for microbial analysis. The focus of the research was on the spores of important air contaminants: Streptomyces albus and Thermoactinomyces vulgaris. For the measurement of the enzymatic activity, the chosen fluorogenic substrate was added to a mixture of spores and nutrient media, and the resulting fluorescence was measured with a spectrofluorometer. Fluorogenic substrates were found to show enzymatic activities even for dormant spores. Comparison of the enzymatic activities of dormant spores with those of vegetative cells showed similarity of the enzymatic profiles but higher activity for vegetative cells. The increase of enzymatic activity from dormant spores to vegetative cells was not linear but fluctuating. The largest fluctuations were found after 4 to 5 h of incubation. The enzymatic activities of S. albus were 10 to 50 times lower than those of T. vulgaris, except for the dehydrogenase activity, which was seven times higher. These results indicate that analysis with fluorogenic substrates has the potential for becoming a fast and sensitive method for the enumeration and identification of airborne actinomycete spores.
机译:为了寻找一种检测放线菌孢子的新方法,研究了七个荧光底物与不同组的酶,酯酶,脂肪酶,磷酸酶和脱氢酶之间的反应。选择荧光测量作为微生物分析的快速而灵敏的方法。研究的重点是重要空气污染物的孢子:白链霉菌和寻常型热放线菌。为了测量酶活性,将所选的荧光底物添加到孢子和营养培养基的混合物中,并使用分光荧光计测量所得的荧光。发现即使对于休眠的孢子,荧光底物也显示出酶活性。休眠孢子的酶活性与营养细胞的酶活性的比较显示了酶学特征的相似性,但营养细胞的活性更高。从休眠孢子到营养细胞的酶促活性的增加不是线性的而是波动的。孵育4至5小时后发现最大的波动。除脱氢酶活性高出七倍以外,白al的酶促活性比寻常红球菌低十到五十倍。这些结果表明,使用荧光底物进行分析有可能成为枚举和鉴定空气中放线菌孢子的一种快速而灵敏的方法。

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