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论文类型：期刊论文

发表时间：2018-11-20

发表刊物：ANALYTICAL CHEMISTRY

收录刊物：SCIE、Scopus

卷号：90

期号：22

页面范围：13341-13347

ISSN号：0003-2700

关键字：Fluorescence; Fluorescence spectroscopy; Glycosylation; Photons; Probes, Bioactive compounds; Fluorescent probes; Glucosyltransferases; Intracellular imaging; Molecular analysis; O-glycosylation; Real-time detection; Robust capability, Fungi

摘要：As is well-known, fungi are an important biocatalysis model of glucosylation and have been widely applied for bioactive compounds glucosylation mediated by the intracellular glucosytransferases (GTs). However, there is no efficient method for the real-time detection of GTs and the rapid isolation of the target fungi strains with the high expression of GTs. In the present work, we first developed a two-photon ratiometric fluorescent probe N-(n-butyl)-4-hydroxy-1,8-naphthalimide (NHN) for detecting the glucosyl-transferases activity and intracellular imaging of GTs. Under UV light (365 nm), the transformed product of NHN mediated by intracellular glucosyltransferase displayed blue emission to guide the rapid isolation of fungal strains possessing overexpression of GTs from complex soil samples. Finally, by using the fluorescent probe, two target fungi were isolated and identified to be Rhizopus oryzae and Mucor circinelloides by molecular analysis, and they exhibited a robust capability for regio- and stereospecific O-glycosylation. Our results fully demonstrated that NHN may be a promising tool for guiding real-time GTs activity in fungal strains and even for developing natural fungal strains with GTs overexpression.