香豆素类荧光底物的合成及其在微生物检测中的应用进展

摘要

香豆素类荧光底物主要是由作为荧光团的羟基香豆素和7-氨基-4-甲基香豆素或者它们的取代物的各类衍生物，包括糖苷类、羧酸酯类、磷酸酯类、肽类等。本文介绍了这些底物的合成方法。荧光团一般通过Pechmann缩合法合成，然后再与乙酰溴代糖、酰氯、亚磷酸酯、叔丁氧基保护的氨基酸等缩合生成相应的荧光底物，像4-甲基伞形酮基-β-D-吡喃半乳糖苷、4-甲基伞形酮辛酸酯和L-丙氨酰-7-氨基-4-甲基伞形酮等。将荧光底物加入到培养基中，可以实现对微生物的快速检测。荧光底物应用于微生物检测的依据是特异性酶反应，不同的荧光底物对应的酶和所检测的微生物不同。当单一底物不能有效检测和鉴定目标微生物时，采用复合底物能明显提高检测效果。此外，本文还指出了现有荧光底物存在的缺点，并对未来发展进行了展望。%Coumarin fluorogenic substrates are mainly derivatives of fluorophore , such as hydroxycoumarin and 7-amino-4-methyl coumarin or their substitution , including glycosides , carboxylic esters,phosphate,peptides and the like. This review introduces the synthesis of these substrates. Fluorophores are generally synthesized by Pechmann condensation,then their condensation with acetyl bromide sugar,acyl chloride,phosphite ester and tert-butoxy-protected amino acids generate the corresponding fluorogenic substrate,like 4-methylum belli feryl-β-D-galactopyranoside, 4-methylumbelliferyl caprylate,L-alanyl-7-amino-4-methylumbelliferone and the like. The fluorescent substrates are added to the media to achieve rapid detection of microorganisms.The application of fluorogenic substrates in microbiological testing is based on specific enzyme reactions. Different fluorescent enzyme substrates correspond to different enzymes which target different microorganisms.When a single substrate can not effectively detect and identify the target microorganisms , the composite substrates can be used to improve the detection results. In addition,problems of the existing fluorogenic substrate in microbial detection are pointed out,and future directions are discussed.