Novel chemical-and protein-mediated methods for glucosamine detection

查看全文 作　　者：Linshu [1]Chen;Pedro [1]Laborda;Zhipeng [1]Cai;Andrew Kevin [1]Hagan;Aimin [2]Lu;Josef [1]Voglmeir;Li [1]Liu 机构地区：[1]Glycomics and Glycan Bioengineering Research Center(GGBRC),College of Food Science and Technology,Nanjing Agricultural University,1 Weigang,Nanjing 210095,People's Republic of China;[2]College of Sciences,Nanjing Agricultural University,1 Weigang,Nanjing 210095,People's Republic of China 出　　处：《Food Materials Research》索引2022年第2卷第1期,共8页 基　　金：supported in parts by the National Natural Science Foundation of China (grant numbers 31471703,31671854,and 31871754);the 100 Foreign Talents Plan (grant number JSB2014012). 摘　　要：We describe two novel approaches for the determination of glucosamine(GlcN).The first approach is based on the chemical derivatization of GlcN with the non-fluorophor 1,3-diphenyl-1,3-propanedione(DPPD),which results in a condensation product with interesting fluorescent properties.The obtained compound was isolated by silica-gel chromatography and its structure elucidated by NMR and mass spectrometry.The second approach is based on a previously undescribed sensitivity of the enzyme glucosamine-6-phosphate deaminase(GPDA)towards GlcN,which resulted in the precipitation of the enzyme.Using a rational enzyme engineering approach and both liquid-based and plate-based screening methods,mutational GPDA variants with significantly improved precipitation properties were identified and characterized.These novel glucosamine detection methods may be a useful addition to the repertoire of currently available glucosamine detection sensors. 关 键 词：PROPERTIES CHEMICAL RATIONAL